bacoside a: role in cigarette smoking induced changes in brain · 2018. 11. 27. · bacoside a:...

Review ArticleBacoside A Role in Cigarette Smoking InducedChanges in Brain

G Vani K Anbarasi and C S Shyamaladevi

Department of Biochemistry University of Madras Guindy Campus Chennai 600 025 India

Correspondence should be addressed to C S Shyamaladevi cssdevigmailcom

Received 19 October 2014 Revised 11 January 2015 Accepted 26 February 2015

Academic Editor Andrea Zangara

Copyright copy 2015 G Vani et al This is an open access article distributed under the Creative Commons Attribution License whichpermits unrestricted use distribution and reproduction in any medium provided the original work is properly cited

Cigarette smoking (CS) is amajor health hazard that exerts diverse physiologic and biochemical effectsmediated by the componentspresent and generated during smoking Recent experimental studies have shown predisposition to several biological consequencesfrom both active and passive cigarette smoke exposure In particular passive smoking is linked to a number of adverse healtheffects which are equally harmful as active smoking A pragmatic approach should be considered for designing a pharmacologicalintervention to combat the adverse effects of passive smoking This review describes the results from a controlled experimentalcondition testing the effect of bacoside A (BA) on the causal role of passivesecondhand smoke exposure that caused pathologicaland neurological changes in rat brain Chronic exposure to cigarette smoke induced significant changes in rat brain histologicallyand at the neurotransmitter level lipid peroxidation states mitochondrial functions membrane alterations and apoptotic damagein rat brain Bacoside A is a neuroactive agent isolated from Bacopamonnieri As a neuroactive agent BAwas effective in combatingthese changes Future research should examine the effects of BA atmolecular level and assess its functional effects onneurobiologicaland behavioral processes associated with passive smoke

1 Introduction

Cigarette smoking is an intractable and preventable publichealth problem It is an important risk factor involved in thepathogenic pathways of a variety of disorders TheWHO hasdeclared global tobacco epidemic and planned ldquoFrameworkConvention for Tobacco Controlrdquo [1] Tobacco smoke isa toxic air contaminant and secondhand tobacco smoke(SHS) is a formidable health hazard [2] Epidemiologicalstudies show increased risk for behavioral and cognitiveproblems and a greater incidence of mental disorders inchildren exposed to environmental tobacco smoke [3ndash5]Prenatal maternal exposure to cigarette smoke has been welldocumented to induce neurological as well as many otherlasting health effects [6 7] Etiological evidences supportthe involvement of neurotransmitter systems oxidative andnitrogen stress mitochondrial dysfunction and neuroge-netic and epigenetic changes in secondhandpassive smokinginduced brain changes and the associated pathways have beenextensively reviewed [8ndash11]

Despite the significant health impacts arising from pas-sive smoking little attention is paid to combat the neurologi-cal changes associated with itThis review critically examinesand summarizes the study made on the neuroprotective roleof BA in rats exposed to passive cigarette smoke and itssequelae with focus on the neurotransmitter systems oxida-tive and lipid peroxidative mitochondrial dysfunction andapoptotic changes in rat brainThese results can be integratedwith other theories in holistically combating passive smokinginduced neurological changes

2 Physical and Biochemical Properties ofCigarette Smoke

Cigarette smoke is divided into two phases a tar phase anda gas phase The tar or particulate phase is defined as thematerial that is trapped when the smoke stream is passedthrough the Cambridge glass-fiber filter that retains 999 ofall particulate material with a size sim01m [12] The gas phaseis the material that passes through the filter The particulate

Hindawi Publishing CorporationEvidence-Based Complementary and Alternative MedicineVolume 2015 Article ID 286137 16 pageshttpdxdoiorg1011552015286137

2 Evidence-Based Complementary and Alternative Medicine

(tar) phase of cigarette smoke contains sim1017 free radicalsgand the gas phase contains sim1015 free radicalspuff [12] Theradicals associated with the tar phase are long-lived (hours tomonths) whereas the radicals associated with the gas phasehave a shorter life span (seconds) [12ndash14]

Cigarette smoke that is drawn through the tobacco intoan active smokerrsquos mouth is known as mainstream smoke(MS) and the smoke emitted from the burning ends of acigarette is the sidestream smoke (SS) Mainstream cigarettesmoke comprises 8 of tar and 92 of gaseous compo-nents [12] Environmental tobacco smoke (ETS) results fromthe combination of sidestream smoke (85) and a smallfraction of exhaled mainstream smoke (15) from smokers[13] Importantly the concentration of numerous toxins isdramatically (up to 100-fold) elevated in SS when comparedwith MS and the complex mixture of toxins is attributedrather to a specific component of cigarette smoke to thepotential adverse impact of passive smoke on health [15]

Aside from specific chemical constituents certain physic-ochemical properties of smoke may participate in diseaseprocesses The pH of the smoke affects the site and degreeof nicotine absorption as well as the smokerrsquos depth ofinhalation The oxidation-reduction state of the smoke isimportant because oxidants influence the maturation ofcholesterol-laden plaques in the coronary arteries and otherblood vessels In short cigarette smoke is farmore than a triadof tar nicotine and carbon monoxide [16]

Although SS and MS smoke have qualitatively similarchemical compositions the respective quantities of individ-ual smoke constituents are different [17] The exposure to SSsmoke depends on the distance from the burning cigaretteand conditions of ventilation the higher concentrations ofcertain toxic and carcinogenic chemicals in SS smoke result inmeasurable levels of these chemicals in nonsmokers exposedto ETS [18]

3 Pharmacodynamics of Smoking

Although most of the toxicity of smoking is related to othercomponents of cigarette smoke it is primarily the pharmaco-logic effects of nicotine that produce the addiction to tobaccoAn understanding of how nicotine produces addiction andinfluences smoking behavior provides a necessary basis foroptimal smoking cessation intervention Cigarette smokecontains 10ndash14mg of nicotine [19] of which 1ndash15mg isabsorbed systemically in the lungs through inhalation [20]Nicotine rapidly enters the pulmonary venous circulationreaches the brain within 10ndash20 s and readily diffuses intobrain tissue [21] and binds to nicotine acetylcholine receptors(nAChRs) [22] Acutely cigarette smoking induces posi-tive reinforcing effects including mild euphoria heightenedarousal reduced appetite and reduced stress anxiety andpain [23]

Nicotine in cigarette smoke is alkaline and readilycrosses the blood brain barrier It mediates the stimulationof mesolimbic dopamine system It is also involved inmodulating other classical neurotransmitters in the brainincluding catecholamines serotonin GABA and glutamate

[24] It induces addiction as it shares many properties ofpsychostimulant drugs such as cocaine and amphetamine[25]With repeated exposure to nicotine toxicity gives way totolerance and addiction Overtime tolerance is accompaniedby increased intake of nicotine to attain the same effectsinitially taken which leads to physical dependence com-pelling higher intake lest it induces withdrawal symptomThis leads to reinforcement mechanisms finally leading toaddiction [26] Substances other than nicotine present incigarette smoke are also involved in mediating the harmfuleffects in nervous system Other tobacco smoke constituentsaffect the structural and functional integrity of blood brainbarrier [27 28]

4 Neuropharmacological Effects ofCigarette Smoking

Cigarette smoking is an important environmental agingaccelerator [29] partly because it induces oxidative stress inmultiple organs including the brain and is presented in manydiseases including cognition-related or neurodegeneration-related pathological changes [30]This should be presented todemonstrate a direct linkage between smoking and cognitiveimpairment

The incidence of cerebrovascular diseases (CVDs)increases with cigarette smoking as cessation of smokingdecreases its incidence [31] Smoking is a modifiablerisk factor for stroke [32] primarily due to hypertension[33] Other neurological diseases for which smoking isa risk factor include hypoxia cerebral ischemia cerebralhemorrhage brain infarction subarachnoid hemorrhageand tardive dyskinesia [34] Neuroleptic Parkinsonismresembling Idiopathic Parkinsonrsquos disease is associated withsmoking as higher doses of nicotine exert an inhibitoryeffect [35] Cerebral symptoms like brain atrophy and ataxiaare exaggerated with smoking [36] Reports also suggestthat cigarette smoking is protective in the development ofParkinsonrsquos disease [37] but is an important risk factor inAlzheimerrsquos disease [38] as it accelerates cognitive declineand dementia [39]

5 Role of Passive Smoking

The evidence that active smoking is a risk factor forcardiovascular disease (CAD) and the leading cause ofpreventable death is overwhelming However exposure topassive cigarette smoke also exerts detrimental effects onvascular homoeostasis [40] Importantlymost of these effectsappear to be characterized by a rapid onset For example therelatively low doses of toxins inhaled by passive smoking aresufficient to elicit acute endothelial dysfunction and theseeffects may be related at least in part to the inactivation ofnitric oxide (NO) Moreover passive smoking may directlyimpair the viability of endothelial cells and reduce thenumber and functional activity of circulating endothelialprogenitor cells In addition platelets of nonsmokers appearto be susceptible to proaggregatory changes with everypassive smoke exposure Overall passive smoke induces

Evidence-Based Complementary and Alternative Medicine 3

oxidative stress and promotes vascular inflammation Apartfrom vasoconstriction and thrombus formation however themyocardial oxygen balance is further impaired by adrenergicstimulation and autonomic dysfunction [41] These datastrongly suggest that passive smoking is capable of precipitat-ing acutemanifestations as it increases the odds of developingcognitive impairment [42] and 3-fold increase in the riskfor dementia causing neurofibrillary changes depictive ofAlzheimerrsquos disease [43]

Hence one generally overlooked factor contributing tothe escalation of tobacco abuse is passive smoking Nicotinefrom secondhand smoke exposure results in an increase inplasma nicotine concentration of sim02 ngmL and amountsto substantial brain 12057241205732 nAChR occupancy (19) in bothsmokers and nonsmokers compared with 087 ngmL and50 12057241205732 nAChR occupancy from actively smoking onecigarette [44 45] Secondhand smoking is clearly linked toserious illnesses among nonsmokers including asthma heartdisease sudden infant death syndrome and cancer [46]

6 Pharmacological Intervention inCigarette Smoking

With the available understanding of the biological effects ofcigarette smoking many treatment strategies are availableThe primary target is nicotine hence nicotine replacementtherapy or nicotine substitution in the form of chewing gumstransdermal patch and spray are employed in smoking ces-sation aid [47ndash49] Other pharmacological therapies includereceptor antagonists and nicotinic antagonists (mecamy-lamine and chlorisondamine) [50] opiate antagonists andnaloxone and naltrexone [51] nonreceptor antagonists [52]serotonin uptake inhibitors (zimelidine and citalopram) andmonoamine oxidase inhibitor [53] and antidepressants [5455] Angiotensin converting enzymes (ACE) inhibitors andcalcium antagonists are also effective in combating smokinginduced toxicity [56] These drugs are mainly indicated toreduce the severity of tobacco withdrawal but not as an aidto stop smoking [57]

Currently the first line therapy for smoking cessationincludes bupropion (amfebutamone) an atypical antide-pressant that inhibits norepinephrine uptake and dopamineuptake [58] The metabolite of bupropion (2S3S) hydrox-ybupropion is an antagonist on the 12057241205732 (nAChR) [59]Another agent is varenicline which is a highly selective par-tial agonist of the 12057241205732 (nAChR) that stimulates dopaminerelease in the nucleus accumbens (nAC) but to a much lessextent than nicotine itself [60]

These interventions are improbable and ineffective inrendering protection against secondhand smoke Due topoor pharmacological management of passive smokingit was hypothesized that intervening with a neuroactiveagent can prevent or minimize neurological changes Studieson supplementation with vitamins E C and A [61 62]antioxidants glutathione N-acetyl cysteine and superoxidedismutase [63] and fish oil curcumin and green tea [64ndash66]have reported to offer protection against smoking induceddamages

In this context this study evaluated the effect of BAan active constituent isolated from Bacopa monnieri againstsmoking induced damages in rat brain Bacopa monnieriexerts neuropharmacological effects [67] and is effective inthe treatment ofmental illness and epilepsy [68] Its biologicaleffects include free radical scavenging [69] vasodilatory [7071] and mast cell stabilizing [72] activities The variousbiological activities of BA have been reviewed in detail [7374]

Bacoside A is 3-(a-L-arabinopyranosyl)-O-b-D-glucopy-ranoside-10 20-dihydroxy-16-keto-dammar-24-ene [75] andis the major chemical entity responsible for neuropharmaco-logical effects and the nootropic action or antiamnestic effectof Bacopa monniera Bacoside A cooccurs with bacoside Bthe latter differing only in optical rotation and is probablyan artefact produced during the process of isolating BA [76]On acid hydrolysis bacosides yield a mixture of aglyconesbacogenins A1 A2 and A3 [77] which are artefacts two gen-uine sapogenins jujubogenin and pseudojujubogenin andbacogeninA4 identified as ebelin lactone pseudojujubogenin[78]

7 Methods

71 Isolation of Bacoside A The plant Bacopa monniera wascollected in and around Chennai India and authenticated byDr P Brindha Central Research Institute (Siddha) ChennaiIndiaThe dammarane type triterpenoid saponin BAwas iso-lated from the plant by the standard procedure The purity ofthe isolated BA was identified by thin layer chromatography(TLC) and infrared (IR) spectrum analysis using standard BA[79]

72 Experimental Setup Adult male albino rats of Wistarstrain (120ndash200 g) were used for the present study The ratswere provided with standard pelleted rat feed and water adlibitum They were acclimatized to the laboratory conditionsand maintained under 12 h light and dark cycles The exper-iments were carried out in accordance with the guidelinesprovided by the Institutional Animal Ethical Committee [79]

The animals were divided into four groups of 6 animalseach Group I control Group II (CS) rats exposed tocigarette smoke Group III (BA) rats administered with BA(10mgkg bwday po) Group IV (CS + BA) rats exposedto cigarette smoke and simultaneously administered with BAGroup II and Group IV rats were exposed to cigarette smokefollowing a standardmethod as described [79] for a period of12 weeks

The rats were exposed to side stream cigarette smoke inwhole body smoke exposure chamber The rats were exposedtwice daily as described [80 81] The experimental periodlasted for 12 weeks Drug control animals received aqueoussuspension of BA in 1 gum acacia orally at a dosage of10mgkg bwday for 12 weeks whereas experimental animalsexposed to cigarette smoke (Scissors Standard Cigarette)were simultaneously administered with BA at the same doseControl animals received a corresponding volume of thevehicle suspended in normal salineThe same brand of locally


Table 1 Constituents of the cigarette smoke

Smoke constituents ConcentrationcigaretteNicotine 18mgCarbon monoxide 20mgTotal particulate matter 32mgAcetaldehyde 09mgHydrogen cyanide 225mgBenzene 38mgNrsquonitrosonorcotine 240mg

available cigarette was used throughout the experiment(Scissors Standard WD amp HOWills Hyderabad DeccanCigarette Factory) Control animals were subjected to thesame handling and time in the smoke exposure chamber withair replacing smokeair mixtureThe composition of cigarettesmoke was analyzed at Tamil Nadu Pollution Control BoardChennai and the constituents present are listed in Table 1

8 Results and Discussion

81 Structural Brain Changes and Clinical CorrelatesCigarette smoking is associated with diverse structuralchanges in brain probably as a consequence of toxicity oras an adaptive response causing a reduction in integrity ofcerebral white matter microstructure [82] and gray mattervolumes [83 84] and these changes appear correlated withthe magnitude of cigarette exposure Smoking inducedstructural changes in brain are associated with cognitivedeficits [85] as well with the integrity of white matter andglial proliferation [86] In gross the microstructural changesin key brain regions and white matter tracts have a negativeimpact in cigarette smokers

In the present study histological changes were preva-lent in brain of rats exposed to cigarette smoke that wereinflammatory and edematous in the cerebrum (Figure 1)Smoking induced inflammatory changes were also markedby increased activity of CK-MB isoenzyme in serum[79] an early marker for pathological changes like cere-bral damage [87] 4-N-Methyl-N-nitrosamino-1-(3-pyridyl)-1-butanone (NNK) is a major nitrosamine present in sub-stantial concentration in MS and SS that causes oxidativestress and triggers neuroinflammation in brain [88 89]Inflammation plays a pivotal role in extremely wide arrayof disease conditions ranging from viral diseases of CNSto neurodegenerative disorders NKK mediated microglialactivation leads to profound increase in inflammatory medi-ators The inflamed milieu may cause neuronal damage [90]A decrease in the inflammatory changes was noted in BAtreated rats exposed to cigarette smoke which could be dueto the anti-inflammatory effect of BA [91] and the reductionin cerebral inflammatory changes in treated rats were alsoreflected in lowered levels of CK-MB as against untreated rats[79]

Electroencephalography (EEG) of rat brain monitoredfrontal and parietal regional changes in brain as electricalchanges as 120572 120573 120575 and 120579 waves Cigarette smoke exposed

rats presented depressed 120575 and increased 120572 waves (Figure 2)A desynchronized and electrically active EEG pattern isnoted in smokers [92] Acute smoking accelerates dominantfrequency fast waves 120572 and 120573 with a reduction in slow wave120575 and 120579 waves illustrate a stimulant action [93] whereaschronic smoking induces less 120572 wave and more 120573 wave [94]In rats treated with BA and exposed to cigarette smoke theEEG pattern was devoid of desynchronization and lackedstimulatory wave an effect also noted among cholinergicagonists mecamylamine and scopolamine This shows theanticholinergic effect of BA and effective against smokinginduced stimulation of brain

82 Neurotransmitter Systems Neurotransmitters mediatediverse pharmacological effects on central and peripheralnervous system and participate in reinforcing mood eleva-tion and cognitive functions [95] A balance in their rate ofsynthesis and utilization constitutes the regulatory mecha-nism in neurotransmission Smokers have positive effects likepleasure arousal and relaxation as well as negative effectslike depression and anxiety The functional antagonismpresented in cigarette smoking is related to desensitizationof nAChR Nicotine in cigarette smoke upregulates nAChR(pre- and postsynaptic) which in turn interacts with thenoradrenergic cannabinoid dopaminergic cholinergic andserotonergic systems [96] and increases the levels of nore-pinephrine dopamine acetylcholine and serotonin [97]

Cigarette smoking upregulates nAChR in the brainincluding the common 12057241205732 nAChR subtype [23] In thepresent study an upregulation of 1205724 subunit was evident inrats exposed to cigarette smoke (Figure 3) Chronic adminis-tration of nicotine also upregulates nAChRs [98 99] causingan increased receptor function and sensitivity to nicotineThis results in increased trafficking of nAChRs to the cellsurface increased receptor assembly andor maturationor other mechanisms [100] In smokers abstinence fromsmoking normalizes the nAChR upregulation to the levels ofnonsmokers [101 102] Similarly commonly used treatmentsfor smoking cessation also decrease 12057241205732 nAChR to nearnormal levels as in nonsmokers In the exploratory analysesdecreases in 12057241205732 nAChR levels are associated with decreasein the perceived rewarding properties of nicotine [103 104]Hence a downregulation of 1205724 nAChR in BA treated smokeexposed rats could be associated with diminished rewardfrom cigarettes (presumablymediated at least in part throughdopamine release) Taken together these findings indicatethat the role of BA on nAChR regulation could be vital inmodulating nicotine response and reward pathway in chroniccigarette smoking However the mechanism on how BAinfluences the upregulation remains to be understood

Nicotine is cholinergic by increasing the release of acetyl-choline (ACh) from axonal stores and inhibits its clearance byinhibiting acetylcholine esterase (AChE) [105ndash107] Increasedaccumulation of ACh increases the electrical activity in ratbrain [107] This accounts for the increase in most of theneurotransmitters in rats exposed to cigarette smoke (Figures4ndash6) In BA treated rats the activities of AChEwere increased


(a)times10)(H amp E

(b)

times40)(H amp E

(H amp E times40)(c) (d)

times40)(H amp E

Figure 1 Sections of rat brain cerebellum (a) Control rats showing normal architecture (b) CS rats showing mild gliosis edema necrosisand Purkinje cell damage (c) BA rats showing normal architecture with no significant changes (d) CS + BA rats showing normalmorphologyof Purkinje cells

120572

120573

120575

120579

Control CS BA CS + BA

Control CS BA


Control

Parietal regions Frontal regions

CS BA CS + BA

CS + BA

Figure 2 Electroencephalographic pattern of frontal and parietalregions of rat brain

(Figure 7) which could have decreased the lowered levels ofACh This confirms the anticholinergic effect of BA [108]

Increases in plasma catecholamines are known to occurwith smoking [109] Upregulation of nAChR increases therelease of catecholamines epinephrine and norepinephrinean effect mediated through the tyrosine hydroxylase activity[110] Vasoconstrictor effects observed in smoking are related

to increases in norepinephrine [111] In the present studysmoking induced an increase in the levels of epinephrineand norepinephrine in rat brain (Figure 8) However BAadministration maintained the levels of norepinephrine intreated rats The observed lowering could be due to thedownregulation of nAChR by BA Apart from its ability toinduce downregulation of nAChR expression BA could haveinteracted with tyrosine hydroxylase [112] andmodulated therelease of catecholamines

Nicotine also influences the release of serotonin and ithas been reported to have a dual role as it induces both anincrease and decrease [113 114] In the present study cigarettesmoking increased the serotonin level in rats Serotonergicdysfunction has also been in smokers [115] Serotonergicdysfunction is associated with clinical depression and depres-sion is far more prevalent among smokers [116] suggestinga possible link Further compounds that increase dopamineand its metabolites concentration have abuse potential likeopiates and cocaine whereas those which lower dopamineinduce cognitive behavioral andmotor coordination defects[117]The role of BA on serotonin [118] could havemaintainedthe levels in treated animals (Figure 8) Physiologically highlevel of neuronal dopamine induces greater oxidative stressderived from dopamine [119]These results confirm the effectof Bacopa monnieri extract in normalizing norepinephrineserotonin and dopamine in cortex and hippocampus of ratsin both acute and chronic unpredictable stress [120] Inthe cigarette smoke exposed rats an increase in dopaminelevels was observed but in BA administered rats the levels


times100

(a)

times100

(b)

times100

(c)times100

(d)

Figure 3 Immunohistochemical analysis of nAChR (1205724) expression in rat brain cerebellum (a) Control rats showing normal expression ofnAChR (b) CS rats showing increased expression of nAChR (c) BA rats showing normal expression of nAChR (d) CS + BA rats showingdecreased expression of nAChR

NS

0

5

10

15

20

25

30

35

Control CS BA

(nm

oles

g ti

ssue

)

CS + BA

F valueACh = 14948

lowast

lowast

Figure 4 Levels of acetylcholine in brain of control and experi-mental animals Values are expressed as Mean plusmn SD Significance isindicated for comparisons between control and CS and BA GroupCS versus CS + BA with Dunnettrsquos T3 post hoc multiple comparisontest lowast119875 lt 0001 NS nonsignificant

were maintained at near normal This reflects the safety andsubsequent tolerability of BA in preclinical models as it didnot induce any untoward and toxic effect

Most of the nicotine-mediated release of neurotrans-mitters occurs via modulation by presynaptic nAChRsalthough direct release of neurotransmitters also occurs [121]

Dopamine release is facilitated by nicotine-mediated aug-mentation of glutamate release and with long term treatmentby the inhibition of GABA release [122] In addition todirect and indirect stimulation of neurotransmitter releasechronic cigarette smoking (but not nicotine administration)reduces brain monoamine oxidases A and B (MAO-A andMAO-B) activity which would be expected to increasemonoaminergic neurotransmitter levels such as dopamineand norepinephrine in synapses thus augmenting the effectsof nicotine and contributing to addiction [123] Inhibition ofMAO facilitates acquisition of nicotine self-administration inrats supporting the idea that MAO inhibition interacts withnicotine to reinforce tobacco dependence [124] Decreasedactivity of MAO in cigarette smoking exposed rats (Figure 8)confirms reports that have shown downregulation of MAOexpression including MAO-A and MAO-B in the brain[125 126] as well as influencing methylation of MAO pro-moter genes [127] This lowering could have resulted in anincrease in dopamine content in cigarette smoke exposedrats Increases inMAO activities in BA treated rats (Figure 8)confirm the reports of recent studies which have shown theinfluences of Bacopamonnieri on the activities ofMAO [128]

Polyamines play a key role in brain cell replicationdifferentiation and regulation of nAChRs and they influencesynaptic transmission [129 130] Alterations in polyaminegating of cholinergic synaptic signaling contribute to adverseneurobehavioral effects of numerous neuroteratogens [130]


NS

NS

NSNS

0

10

20

30

40

50

60

Control CS BA

(ng

g tis

sue)

Norepinephrine cortex Norepinephrine hippocampusEpinephrine cortex Epinephrine hippocampus

Norepinephrine

Epinephrine

CS + BA

F value

lowast

lowast

lowastlowast

lowast

lowast

lowast lowast

Cortex = 1121Hippocampus = 3203


Figure 5 Levels of norepinephrine and epinephrine in brain ofcontrol and experimental animals Values are expressed as Mean plusmnSD Significance is indicated for comparisons between control andCS and BA Group CS versus CS + BA with Dunnettrsquos T3 post hocmultiple comparison test lowast119875 lt 0001 NS nonsignificant

NS

NS

NSNS

0

10

20

30

40

50

60

Control CS BA

(ng

g tis

sue)

Dopamine hypothalamus Dopamine striatumSerotonin hypothalamus Serotonin striatum

lowast

lowastlowast

lowast

lowast

lowast

lowast

lowast

CS + BA

Figure 6 Levels of dopamine and serotonin in brain of controland experimental animals Values are expressed as Mean plusmn SDSignificance is indicated for comparisons between control and CSand BA CS versus CS + BA with Dunnettrsquos T3 post hoc multiplecomparison test lowast119875 lt 0001 NS nonsignificant

Ornithine decarboxylase (ODC) is the rate limiting enzymein the maintenance of polyamine levels Inhibition of ODCinhibits growth and induces gross dysmorphology upreg-ulating the 1205727 and 12057241205732 nAChR This is accompaniedby abnormalities in macromolecular indices of cell packingdensity and cell membrane surface area In chronic cigarettesmoking exposed rats ODC activity increased significantly(Table 2)

Excitotoxic challenge induces neuronal proliferation andinduces ODC [131] Induction of ODC is neuroprotectivein cerebral ischemia [132] and however is also a commonresponse in various pathological stimuli in brain such as

NS

0100200300400500600700800900

Control CS BA

min

mg

prot

ein)

(120583

mol

es o

f thi

ocho

line l

iber

ated

CS + BA

F valueAChE = 1015

lowast

lowast

Figure 7 Activities of acetylcholine esterase (AChE) in brain ofcontrol and experimental animals Significance is indicated forcomparisons between control and CS and BA CS versus CS + BAwith Dunnettrsquos T3 post hoc multiple comparison test lowast119875 lt 0001NS nonsignificant

NS

0010203040506070809

Control CS BA

liber

ated

min

mg

prot

ein)

(120583m

oles

of b

enzla

dehy

de

CS + BA

F valueMAO = 1015

lowast

lowast

Figure 8 Activities of monoamine oxidase (MAO) in brain ofcontrol and experimental animals Significance is indicated forcomparisons between control and CS and BA Group CS versusCS + BA with Dunnettrsquos T3 post hoc multiple comparison testlowast119875 lt 0001 NS nonsignificant

physical chemical thermal and metabolic injuries [133] Arelatively long lasting increase in ODC and consequently itsproduct putrescine are causally related to neurodegeneration[134] In the present study cigarette smoking increased theactivities ofODCBA treated rats recorded a decrease inODCactivity confirming its role in inhibiting neurodegenerativeprocess following cigarette smoking induced excitotoxicity inbrain

83 Nicotine and Cotinine Levels Cigarette smokingincreases the levels of nicotine and its metabolite cotinine topharmacologically active concentrations that are responsiblefor mediating the aspects of nicotine dependence In ratsexposed to cigarette smoke accumulation of cotinine inbrain was noted (Table 3) and the levels were lowered in BAtreated rats The decrease in the levels could have probablyresulted from the increased clearance of cotinine by the


Table 2 Levels of nicotine and cotinine in brain of control and experimental animals Values are expressed as Mean plusmn SD

Parameter Control Cigarette smoke(CS)

Bacoside A(BA)

Cigarette smoke + bacoside A(CS + BA) 119865 value

Nicotine (nggtissue) nd 180 plusmn 12 nd 89 plusmn 5 87008

Cotinine (nggtissue) nd 210 plusmn 15 nd 120 plusmn 8 71814

Significance is indicated for comparisons between control and CS and BA CS versus CS + BAwith Dunnettrsquos T3 post hocmultiple comparison test lowast119875 lt 0001nd not detected

CYP system Although Bacopa monnieri extract reportedlyinhibits CYP enzymes [135] increased clearance of cotinineas noted from a decrease in cotinine levels in BA treated ratsconfirms that purified bacosides do not inhibit CYP insteadthe constituents in crude extract exert an inhibitory effect[136 137]

Cigarette smoking accelerates the metabolism of drugsespecially the ones primarily metabolized by CYP1A2 [138]It delays the clearance of nicotine [139] In smokers nicotineclearance is increased by 14 in 4-day smoking abstinenceand by 36 higher in 7-day smoking abstinence comparedto overnight abstinence Apart from nicotine substance(s)in cigarette smoke as yet unidentified also affect themetabolism of nicotine For instance cotinine slows themetabolism of nicotine since both are metabolized by thesame enzyme [140] However carbon monoxide in cigarettesmoke has no effect on nicotine and cotinine clearance[141] but 120573-nicotyrine a minor alkaloid in cigarette smokeeffectively inhibits CYP2A6 in vitro [142] Thus reducednicotine clearance may also result from downregulation ofCYP expression and not inhibition [143]

Cigarette smoking also induces glucuronidation ofsome drugs such as propranolol and oxazepam andUGT1A9 is the inducible component of 31015840-hydroxycotinineO-glucuronidation [143] Excretion of 31015840-hydroxycotinineO-glucuronide is induced by smoking but the extent ofnicotine and cotinine N-glucuronidation is not significantlyaffected In rats exposed to cigarette smoke increase inUDP-GT was noted and the activities remained unalteredin BA treated rats [144] The adaptogenic role of Bacopamonnieri is evident from increased cotinine clearance [145]

84 Oxidative and Peroxidative Changes Free radicals medi-ated oxidative stress has been implicated in the pathogenesisof smoking-related diseases and antioxidant nutrients arereported to prevent the oxidative damage induced by smok-ing Cigarette smoking modulates antioxidant status in vari-ous organs by increasing lipid peroxidation and prooxidativestate [146] Increased basal and induced lipid peroxidationwere observed in cigarette smoke exposed rat brain [147]Acute exposure to cigarette smoke enhances the productionof antioxidant enzymes as a result of adaptive response thatmitigates the damage [148] but chronic exposure decreasesthe inherent antioxidant defense in brain [149 150]

The constituents of cigarette smoke affect the individualcellular antioxidants differently The quinonesemiquinoneradicals from the tar phase of cigarette smoke inactivate

superoxide dismutase [151] and inhibit catalase in brain [152]Acetaldehyde a major aldehyde from the smoke depletes cellof cellular glutathione [153]Other cellular antioxidants toco-pherols carotenoids and retinol are destructed by cigarettesmoke [154]

Further the cigarette tar contains large amounts ofmetalscomplexed to some components of tar such as odiphenols[155] which can mobilize reactive iron from ferritin andcopper from copper binding protein inducing damage tobrain [156] The heavy metal cadmium in cigarette smokedecreases the bioavailability of selenium (Se) and zinc (Zn)and thus depletes the antioxidant status [157] The role of BAas chelator of transition metal inhibition of free radicals andtermination of lipid peroxidation at the initiation level itself[69] accounts for its protection in cigarette smoke inducedlipid peroxidative damage and combative against oxidativedamage

85 Mitochondrial Functions Mitochondria are the site ofcellular oxidation and provide ATP for various metabolicprocesses and hence are vulnerable to free radical attackMitochondrial damage is prevalent in both heart and brainfollowing cigarette smoke exposure [158 159] Exposure tocigarettes can lead to mitochondrial dysfunction as demon-strated by increased levels of cholesterol lipid peroxides andincreased cholesterolphospholipid ratio in conjunctionwithdecreasedmitochondrial enzymes in rats exposed to cigarettesmoke [160] Chronic cigarette smoking prevented exercise-induced improvement in brain mitochondrial function andneurotransmission [161] Perturbedmitochondrial energeticsis critical in normal brain development [6 162] Cerebellarperturbation can broadly impact regulation of behavioral andcognitive domains [163]

Aerobic demands increase postnatally with heightedsynaptic development requiring more ATP to maintainmembrane polarity Exposure to cigarette smoke perturbedthemitochondria and associated aerobic pathwaysThe effectof BA in regulating the key aerobic ATP production probablyby preventing the peroxidative changes in mitochondriacould be crucial in mitochondrial mediated neurotransmis-sion pathways Brain energetics is highly regulated processand further studies in themechanistics can provide an insightinto the role of BA

86 Membrane Integrity and Electrolyte Balance Derange-ment of membrane bound enzymes and modifications oflipid bilayer alterations following cigarette smoke exposure


Table 3 Activities of ornithine decarboxylase in brain of control and experimental animals Values are expressed as Mean plusmn SD


Bacoside A(BA)


Ornithine decarboxylasenM of 14CO2 releasedhrgtissue

20 plusmn 012 565 plusmn 052 223 plusmn 022 245 plusmn 0023 3489

Significance is indicated for comparisons between control and CS and BA Group CS versus CS + BA with Dunnettrsquos T3 post hoc multiple comparison testlowast119875 lt 0001

nd not detected

resulted in significant decrease in the activities of ATPases[164] Free radicals in cigarette smoke deplete cell proteinsulfhydryl groups and increase in protein carbonyl formation[165] and so does acetaldehyde in cigarette smoke [166]Membrane bound ATPases are thiol-dependent enzymesand modification of thiol groups within the active sites ofthese enzymes lowers their activities in cigarette smoke ratsThe antioxidant role of BA prevented the membrane damageand restored the activities of ATPases Also the restitutionof ATP levels by altering the mitochondrial dysfunctionmaintained the activities of ATPases

Inhibition of Na+K+-ATPase and elevation of Na+ inchronic exposure to cigarette smoke are attributed to theincreased cholesterolphospholipid ratio [167] followed byneuronal apoptotic death mediated by intracellular depletionof K+ and accumulation of Na+ and Ca2+ [168] Plasmamem-brane Ca2+-ATPase (PMCA) is a regulator of intracellularcalcium which undergoes early developmental changes inrat brain as a function of its maturity [169] PMCA is verysensitive to the inhibitory effect of reactive oxygen species(ROS) due to the age dependent oxidative modification ofPMCA and the related chronic oxidative stress [170]

In addition to generation of free radicals cellular degen-eration that is involved in cigarette smoking is related tothe accumulation of advanced glycosylation end-products(AGE) Activities of several enzymes are inhibited due toenzyme protein glycation [171 172] The changes in the Ca2+ATPase can be related to the increased glycation found incigarette smoke exposed rats that in turn may lead to theenzyme protein glycation [173] Alterations in the capacity tomaintain normal calcium homeostasis underlies the reducedcellular function bound with the aging process In thebrain multiple methionines within the calmodulin moleculebecome oxidized to methionine sulfoxides resulting in aninability to activate a range of target proteins includingplasma membrane Ca2+-ATPase [174]

Mg2+-ATPase is not uniformly distributed and differs inrespect to affinity forATP in rat brain regions [175] and is acti-vated by millimolar concentrations of Mg2+ Comparison ofNa+ K+-ATPase and Mg2+-ATPase activities in the synapticplasma membrane from various regions of rat brain revealsthat moderate hypoxia increases the activity of synaptosomalMg2+-ATPase whereas activities of both Ca2+- ATPase andNa+ K+-ATPase are decreased [176]

Increased concentrations of Ca2+ by stimulatingNa+Ca2+ exchanger produce cellular Mg2+ depletion sinceexcessive calcium displaces magnesium from its binding

sites [177] Decrease in Mg2+ in turn inhibits Na+K+-ATPase further as ATP-Mg complex is the actual substratefor the enzyme [178] Rats exposed to cigarette smokeshowed a decrease in the activity of brain Mg2+-ATPaseThe restoration of membrane bound ATPases maintainedthe electrolyte homeostasis in brain impairing electrolytebalance in cigarette smoking

87 Apoptotic and Neurogenic Changes Dysregulation ofapoptosis is an important factor in the pathogenesis ofcigarette smoking [179] Nicotine is involved in both stim-ulation and inhibition of neuronal apoptosis [180ndash182]Apoptosis is suggested as a possible contributing factor inthe pathogenesis of smoking-induced toxicity Exposure tocigarette smoke induced apoptosis as characterized by DNAladdering increased TUNEL-positive cells and apoptoticfeatures evident ultrasctructurally in the brain Adminis-tration of BA prevented expression of hsp70 and neuronalapoptosis during cigarette smoking [183] Extract of BMreduced oxidative stress by improving Nrf2 expression andresults in improvement in antiapoptotic (Bcl2) expressionand decreased proapoptotic (Bax and caspase-3 activity)indicating neuroprotection [184]

88 Therapeutic Implications of BA in Passive Smoking Aninsight into these observations supports the role of BA asa supplement for secondhand smoking Its role on nAChRexpression may underpin its effect on cigarettes inducedneurochemical alteration Generally antidepressants are non-competitive inhibitors of nAChRs [185] and so it is possiblethat the role of BA as a noncompetitive inhibitor to nAChRscould potentially help in controlling the nAChR mediatedupregulation of neurotransmitters and nicotine dependence[186] apart from its role on nAChR expression at thetranscriptional level

Other potential sites of action for BAworthy for consider-ation include its ability to control inflammation and oxidativestress Antioxidants and anti-inflammatory drugs potentiallynegate the anxiolytic behaviors [187 188] a feature alsoprevalent in passive smokers Exploitation of the antioxidantproperty of BA could aid in overcoming oxidative anxietydisorders

9 Conclusion

A number of admonitions exist in the data presented Theinterpretations are drawn from a study involving chronic


exposure of rats to cigarette smoke and not acute cigarettesmoke The cross-sectional nature of this work is hamperedfrom conclusions not drawn from molecular pathwaysFuture research efforts in this area should attempt to addressthese shortcomings It would be useful to ascertain theeffects of BA on individual components of cigarette smokeconstituents involving multiple pathways Given that passivesmoking affects multiple pathways and may increase risk ofdeveloping anxiety triangulation of potential effects involv-ing a combination of animal and humanmodels will likely berequired As the role of BA appears to be multifaceted it mayrepresent a future therapeutic means for secondary smoke Inaddition to its neuroactive role BA as an anti-inflammatoryand antioxidant agent may assist in improving the symptomsas they may do in other conditions pertaining to oxidativestress Further studies addressing this area may elicit insightsinto new therapeutic opportunities

Conflict of Interests

The authors declare that there is no conflict of interestsregarding the publication of this paper

Acknowledgment

This work was supported by the Council for Scientific andIndustrial Research New Delhi

References

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[2] California Environmental Protection Agency ProposedIdentification of Environmental Tobacco Smoke as a Toxic AirContaminant 2005 httprepositoriescdliborgtcsurveysCALEPA2005

[3] L Anderko J Braun and P Auinger ldquoContribution of tobaccosmoke exposure to learning disabilitiesrdquo Journal of ObstetricGynecologic and Neonatal Nursing vol 39 no 1 pp 111ndash1172010

[4] F C Bandiera A Kalaydjian Richardson D J Lee J-P He andK R Merikangas ldquoSecondhand smoke exposure and mentalhealth among children and adolescentsrdquo Archives of Pediatricsand Adolescent Medicine vol 165 no 4 pp 332ndash338 2011

[5] Z Kabir G N Connolly and H R Alpert ldquoSecondhand smokeexposure and neurobehavioral disorders among children in theUnited Statesrdquo Pediatrics vol 128 no 2 pp 263ndash270 2011

[6] S P Doherty J Grabowski C Hoffman S P Ng and JT Zelikoff ldquoEarly life insult from cigarette smoke may bepredictive of chronic diseases later in liferdquo Biomarkers vol 14supplement 1 pp 97ndash101 2009

[7] J R Pauly and T A Slotkin ldquoMaternal tobacco smoking nico-tine replacement and neurobehavioural developmentrdquo ActaPaediatrica International Journal of Paediatrics vol 97 no 10pp 1331ndash1337 2008

[8] M Berk F Kapczinski A C Andreazza et al ldquoPathwaysunderlying neuroprogression in bipolar disorder focus oninflammation oxidative stress and neurotrophic factorsrdquo Neu-roscience and Biobehavioral Reviews vol 35 no 3 pp 804ndash8172011

[9] S Moylan M Maes N R Wray and M Berk ldquoThe neuro-progressive nature of major depressive disorder pathways todisease evolution and resistance and therapeutic implicationsrdquoMolecular Psychiatry vol 18 no 5 pp 595ndash606 2013

[10] K J Ameringer and A M Leventhal ldquoApplying the tripartitemodel of anxiety and depression to cigarette smoking anintegrative reviewrdquo Nicotine and Tobacco Research vol 12 no12 pp 1183ndash1194 2010

[11] O N Niedermaier M L Smith L A Beightol Z Zukowska-Grojec D S Goldstein and D L Eckberg ldquoInfluence ofcigarette smoking on human autonomic functionrdquo Circulationvol 88 no 2 pp 562ndash571 1993

[12] W A Pryor K Stone C E Cross L Machlin and L PackerldquoOxidants in cigarette smoke radicals hydrogen peroxide per-oxynitrate and peroxynitriterdquo Annals of the New York Academyof Sciences vol 686 pp 12ndash28 1993

[13] C J Smith and T H Fischer ldquoParticulate and vapor phase con-stituents of cigarette mainstream smoke and risk of myocardialinfarctionrdquo Atherosclerosis vol 158 no 2 pp 257ndash267 2001

[14] W A Pryor K Stone L-Y Zang and E Bermudez ldquoFraction-ation of aqueous cigarette tar extracts fractions that containthe tar radical cause DNA damagerdquo Chemical Research inToxicology vol 11 no 5 pp 441ndash448 1998

[15] A E Taylor D C Johnson and H Kazemi ldquoEnvironmentaltobacco smoke and cardiovascular disease a position paperfrom the council on cardiopulmonary and critical care Ameri-can Heart Associationrdquo Circulation vol 86 no 2 pp 699ndash7021992

[16] J E Harris ldquoCigarette smoke components and disease cigarettesmoke is morethan a triad of tar nicotine and carbon monox-iderdquo httpcancercontrolcancergovbrpTCRBmonographs7m7 5

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[18] M S Jaakkola and J J K Jaakkola ldquoAssessment of exposure toenvironmental tobacco smokerdquo European Respiratory Journalvol 10 no 10 pp 2384ndash2397 1997

[19] L T Kozlowski N Y Mehta C T Sweeney et al ldquoFilterventilation and nicotine content of tobacco in cigarettes fromCanada the United Kingdom and the United Statesrdquo TobaccoControl vol 7 no 4 pp 369ndash375 1998

[20] A K Armitage C T Dollery C F George T H HousemanP J Lewis and D M Turner ldquoAbsorption and metabolism ofnicotine from cigarettesrdquo British Medical Journal vol 4 no5992 pp 313ndash316 1975

[21] A Cohen and O George ldquoAnimal models of nicotine exposurerelevance to second-hand smoking electronic cigarette use andcompulsive smokingrdquo Frontiers in Psychiatry vol 4 article 412013

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[24] A L Brody ldquoFunctional brain imaging of tobacco use anddependencerdquo Journal of Psychiatric Research vol 40 no 5 pp404ndash418 2006


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[26] I P Stolerman andM J Jarvis ldquoThe scientific case that nicotineis addictiverdquo Psychopharmacology vol 117 no 1 pp 2ndash10 1995

[27] T J Abbruscato S P Lopez K S Mark B T Hawkins and T PDavis ldquoNicotine and cotinine modulate cerebral microvascularpermeability and protein expression of ZO-1 through nicotinicacetylcholine receptors expressed on brain endothelial cellsrdquoJournal of Pharmaceutical Sciences vol 91 no 12 pp 2525ndash25382002

[28] J R Paulson K E Roder G McAfee D D Allen C J VanDer Schyf and T J Abbruscato ldquoTobacco smoke chemicalsattenuate brain-to-blood potassium transport mediated by theNaK2Cl-cotransporter during hypoxia-reoxygenationrdquo Jour-nal of Pharmacology and ExperimentalTherapeutics vol 316 no1 pp 248ndash254 2006

[29] D Bernhard C Moser A Backovic and G Wick ldquoCigarettesmokemdashan aging acceleratorrdquo Experimental Gerontology vol42 no 3 pp 160ndash165 2007

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[31] P Mazzone W Tierney M Hossain V Puvenna D Janigroand L Cucullo ldquoPathophysiological impact of cigarette smokeexposure on the cerebrovascular system with a focus on theblood-brain barrier expanding the awareness of smokingtoxicity in an underappreciated areardquo International Journal ofEnvironmental Research and Public Health vol 7 no 12 pp4111ndash4126 2010

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[36] J A Johnsen and V T Miller ldquoTobacco intolerance on multiplesystem atrophyrdquo Neurology vol 36 no 7 pp 986ndash988 1986

[37] G D Mellick ldquoCYP450 genetics and Parkinsonrsquos disease Genex environment interactions hold the keyrdquo Journal of NeuralTransmission Supplementum no 70 pp 159ndash165 2006

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4120573

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[57] P Pentel and D Malin ldquoA vaccine for nicotine dependencetargeting the drug rather than the brainrdquo Respiration vol 69no 3 pp 193ndash197 2002

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[79] K Anbarasi G Vani K Balakrishna and C S S DevildquoCreatine kinase isoenzyme patterns upon chronic exposureto cigarette smoke protective effect of Bacoside Ardquo VascularPharmacology vol 42 no 2 pp 57ndash61 2005

[80] E Madhukumar and P L Vijayammal ldquoInfluence of cigarettesmoke on cross-linking of dermal collagenrdquo Indian Journal ofExperimental Biology vol 35 no 5 pp 483ndash486 1997

[81] WorldHealthOrganizationWHOReport on the Global TobaccoEpidemic 2011 Warning about the Dangers of Tobacco WorldHealth Organization Geneva Switzerland 2011

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[84] X Zhang E A Stein and L E Hong ldquoSmoking andschizophrenia independently and additively reduce white mat-ter integrity between striatum and frontal cortexrdquo BiologicalPsychiatry vol 68 no 7 pp 674ndash677 2010

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[89] Z Jin F Gao T Flagg and X Deng ldquoTobacco-specificnitrosamine 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanonepromotes functional cooperation of Bcl2 and c-Myc throughphosphorylation in regulating cell survival and proliferationrdquoThe Journal of Biological Chemistry vol 279 no 38 pp40209ndash40219 2004

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[99] X Zhang J-Y Tian A-L Svensson Z-H Gong B Meyersonand A Nordberg ldquoChronic treatments with tacrine and (minus)-nicotine induce different changes of nicotinic and muscarinicacetylcholine receptors in the brain of aged ratrdquo Journal ofNeural Transmission vol 109 no 3 pp 377ndash392 2002

[100] P J Whiting and J M Lindstrom ldquoCharacterization of bovineand human neuronal nicotinic acetylcholine receptors usingmonoclonal antibodiesrdquo Journal of Neuroscience vol 8 no 9pp 3395ndash3404 1988

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[102] K P Cosgrove I Esterlis S A McKee et al ldquoSex differences inavailability of 1205732lowast-nicotinic acetylcholine receptors in recentlyabstinent tobacco smokersrdquo Archives of General Psychiatry vol69 no 4 pp 418ndash427 2012

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and relevance to nicotine addictionrdquo Biochemical Pharmacol-ogy vol 78 no 7 pp 756ndash765 2009

[104] A W Bergen H S Javitz R Krasnow et al ldquoNicotinic acetyl-choline receptor variation and response to smoking cessationtherapiesrdquo Pharmacogenetics and Genomics vol 23 no 2 pp94ndash103 2013

[105] W B Pickworth R M Keenan and J E Henningfield ldquoNico-tine effects and mechanismrdquo in Handbook of NeurotoxicologyL W Chang and R S Dyer Eds pp 808ndash818 Marcel DekkerNew York NY USA 1995

[106] A K Armitage and G H Hall ldquoMode of action of intravenousnicotine in causing a fall of blood pressure in the catrdquo EuropeanJournal of Pharmacology vol 7 no 1 pp 23ndash30 1969

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[109] J H Markovitz L Tolbert and S E Winders ldquoIncreasedserotonin receptor density and platelet GPIIbIIIa activationamong smokersrdquo Arteriosclerosis Thrombosis and VascularBiology vol 19 no 3 pp 762ndash766 1999

[110] S N Mitchell K M Smith M H Joseph and J A GrayldquoIncreases in tyrosine hydroxylase messenger RNA in the locuscoeruleus after a single dose of nicotine are followed by time-dependent increases in enzyme activity and noradrenalinereleaserdquo Neuroscience vol 56 no 4 pp 989ndash997 1993

[111] W G Mayhan ldquoAcute infusion of nicotine potentiatesnorepinephrine-induced vasoconstriction in the hamstercheek pouchrdquo Journal of Laboratory and Clinical Medicine vol133 no 1 pp 48ndash54 1999

[112] D M Rajathei J Preethi H K Singh and K E RajanldquoMolecular docking of bacosides with tryptophan hydroxylasea model to understand the bacosides mechanismrdquo NaturalProducts and Bioprospecting vol 4 no 4 pp 251ndash255 2014

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[114] M Reuben and P B S Clarke ldquoNicotine-evoked [3H] 5-hydroxytryptamine release from rat striatal synaptosomesrdquoNeuropharmacology vol 39 no 2 pp 290ndash299 2000

[115] M E M Benwell D J K Balfour and J M AndersonldquoSmoking-associated changes in the serotonergic systems ofdiscrete regions of human brainrdquo Psychopharmacology vol 102no 1 pp 68ndash72 1990

[116] W Z Potter and H K Manji ldquoCatecholamines in depressionan updaterdquo Clinical Chemistry vol 40 no 2 pp 279ndash287 1994

[117] R F Anda D F Williamson L G Escobedo E E Mast G AGiovino and P L Remington ldquoDepression and the dynamics ofsmoking A national perspectiverdquo The Journal of the AmericanMedical Association vol 264 no 12 pp 1541ndash1545 1990

[118] K E Rajan H K Singh A Parkavi and P D Charles ldquoAttenu-ation of 1-(m-chlorophenyl)-biguanide induced hippocampus-dependent memory impairment by a standardised extract ofBacopa monniera (BESEB CDRI-08)rdquo Neurochemical Researchvol 36 no 11 pp 2136ndash2144 2011

[119] T G Hastings D A Lewis and M J Zigmond ldquoRole ofoxidation in the neurotoxic effects of intrastriatal dopamineinjectionsrdquo Proceedings of the National Academy of Sciences ofthe United States of America vol 93 no 5 pp 1956ndash1961 1996


[120] N Sheikh A Ahmad K B Siripurapu V K Kuchib-hotla S Singh and G Palit ldquoEffect of Bacopa monniera onstress induced changes in plasma corticosterone and brainmonoamines in ratsrdquo Journal of Ethnopharmacology vol 111 no3 pp 671ndash676 2007

[121] S Wonnacott ldquoPresynaptic nicotinic ACh receptorsrdquo Trends inNeurosciences vol 20 no 2 pp 92ndash98 1997

[122] H D Mansvelder and D S McGehee ldquoCellular and synapticmechanisms of nicotine addictionrdquo Journal of Neurobiology vol53 no 4 pp 606ndash617 2002

[123] A Lewis J H Miller and R A Lea ldquoMonoamine oxidase andtobacco dependencerdquo NeuroToxicology vol 28 no 1 pp 182ndash195 2007

[124] A-S Villegier S Lotfipour S C McQuown J D Belluzzi andF M Leslie ldquoTranylcypromine enhancement of nicotine self-administrationrdquo Neuropharmacology vol 52 no 6 pp 1415ndash1425 2007

[125] J S Fowler N D Volkow G-J Wang et al ldquoBrain monoamineoxidase A inhibition in cigarette smokersrdquo Proceedings of theNational Academy of Sciences of the United States of Americavol 93 no 24 pp 14065ndash14069 1996

[126] J S Fowler N D Volkow G-J Wang et al ldquoInhibition ofmonoamine oxidase B in the brains of smokersrdquo Nature vol379 no 6567 pp 733ndash736 1996

[127] F Rendu K Peocrsquoh I Berlin D Thomas and J-M LaunayldquoSmoking related diseases the central role of monoamineoxidaserdquo International Journal of Environmental Research andPublic Health vol 8 no 1 pp 136ndash147 2011

[128] R Singh R Ramakrishna M Bhateria and R S BhattaldquoIn vitro evaluation of Bacopa monniera extract and individ-ual constituents on human recombinant monoamine oxidaseenzymesrdquo Phytotherapy Research vol 28 no 9 pp 1419ndash14222014

[129] C L Law P C L Wong andW F Fong ldquoEffects of polyamineson the uptake of neurotransmitters by rat brain synaptosomesrdquoJournal of Neurochemistry vol 42 no 3 pp 870ndash872 1984

[130] T A Slotkin B D Freibaum C A Tate et al ldquoLong-lasting CNS effects of a short-term chemical knockout ofornithine decarboxylase during development nicotinic cholin-ergic receptor upregulation and subtlemacromolecular changesin adulthoodrdquoBrain Research vol 981 no 1-2 pp 118ndash125 2003

[131] L J Reed and J de Belleroche ldquoInduction of ornithine decar-boxylase in cerebral cortex by excitotoxin lesion of nucleusbasalis association with postsynaptic responsiveness and N-methyl-D-aspartate receptor activationrdquo Journal of Neurochem-istry vol 55 no 3 pp 780ndash787 1990

[132] J A Lukkarinen R A Kauppinen O H Grohn et alldquoNeuroprotective role of ornithine decarboxylase activationin transient focal cerebral ischaemia a study using ornithinedecarboxylase-overexpressing transgenic ratsrdquo European Jour-nal of Neuroscience vol 10 no 6 pp 2046ndash2055 1998

[133] G A Dienel and N F Cruz ldquoInduction of brain ornithinedecarboxylase during recovery from metabolic mechanicalthermal or chemical injuryrdquo Journal of Neurochemistry vol 42no 4 pp 1053ndash1061 1984

[134] F Facchinetti M Virgili P Migani O Barnabei and AContestabile ldquoInduction of brain ornithine decarboxylase aftersystemic or intrastriatal administration of kainic acidrdquo Neuro-science Letters vol 140 no 1 pp 59ndash62 1992

[135] S Ramasamy L V Kiew and L Y Chung ldquoInhibition of humancytochrome P450 enzymes by Bacopa monnieri standardized

extract and constituentsrdquo Molecules vol 19 no 2 pp 2588ndash2601 2014

[136] D Kar Chowdhuri D Parmar P Kakkar R Shukla P K Sethand R C Srimal ldquoAntistress effects of bacosides of Bacopamonnieri modulation of Hsp70 expression superoxide dismu-tase and cytochrome P450 activity in rat brainrdquo PhytotherapyResearch vol 16 no 7 pp 639ndash645 2002

[137] S Zevin and N L Benowitz ldquoDrug interactions with tobaccosmoking An updaterdquo Clinical Pharmacokinetics vol 36 no 6pp 425ndash438 1999

[138] N L Benowitz and P Jacob III ldquoEffects of cigarette smokingand carbon monoxide on nicotine and cotinine metabolismrdquoClinical Pharmacology and Therapeutics vol 67 no 6 pp 653ndash659 2000

[139] N L Benowitz F Kuyt and P Jacob III ldquoCircadian bloodnicotine concentrations during cigarette smokingrdquo ClinicalPharmacology andTherapeutics vol 32 no 6 pp 758ndash764 1982

[140] S Zevin P Jacob III and N Benowitz ldquoCotinine effects onnicotine metabolismrdquo Clinical Pharmacology and Therapeuticsvol 61 no 6 pp 649ndash654 1997

[141] T T Denton X Zhang and J R Cashman ldquoNicotine-relatedalkaloids and metabolites as inhibitors of human cytochromeP-450 2A6rdquo Biochemical Pharmacology vol 67 no 4 pp 751ndash756 2004

[142] K A Schoedel E M Sellers R Palmour and R F Tyn-dale ldquoDown-regulation of hepatic nicotine metabolism and aCYP2A6-like enzyme inAfrican greenmonkeys after long-termnicotine administrationrdquo Molecular Pharmacology vol 63 no1 pp 96ndash104 2003

[143] H L Liston J S Markowitz and C L DeVane ldquoDrugglucuronidation in clinical psychopharmacologyrdquo Journal ofClinical Psychopharmacology vol 21 no 5 pp 500ndash515 2001

[144] K Anbarasi Neuroprotective role of Bacoside A in rats exposedto cigarette smoke [PhD thesis] University ofMadras ChennaiIndia 2005

[145] D Rai G Bhatia G Palit R Pal S Singh and H K SinghldquoAdaptogenic effect of Bacopa monniera (Brahmi)rdquo Pharmacol-ogy Biochemistry and Behavior vol 75 no 4 pp 823ndash830 2003

[146] N Delibas R Ozcankaya I Altuntas and R Sutcu ldquoEffectof cigarette smoke on lipid peroxidation antioxidant enzymesand NMDA receptor subunits 2A and 2B concentration in rathippocampusrdquo Cell Biochemistry and Function vol 21 no 1 pp69ndash73 2003

[147] K Anbarasi G Vani K Balakrishna and C S S Devi ldquoEffectof bacoside A on brain antioxidant status in cigarette smokeexposed ratsrdquo Life Sciences vol 78 no 12 pp 1378ndash1384 2006

[148] J Hilbert and V Mohsenin ldquoAdaptation of lung antioxidants tocigarette smoking in humansrdquoChest vol 110 no 4 pp 916ndash9201996

[149] S A Hulea R Olinescu S Nita D Crocnan and F AKummerow ldquoCigarette smoking causes biochemical changesin blood that are suggestive of oxidative stress a case-controlstudyrdquo Journal of Environmental Pathology Toxicology andOncology vol 14 no 3-4 pp 173ndash180 1995

[150] B Frei T M Forte B N Ames and C E Cross ldquoGasphase oxidants of cigarette smoke induce lipid peroxidationand changes in lipoprotein properties in human blood plasmardquoBiochemical Journal vol 277 no 1 pp 133ndash138 1991

[151] G R Duthie and J R Arthur ldquoCigarette smoking as an inducerof oxidative stressrdquo in Exercise and Oxygen Toxicity K SChandan L Packer and H Osmo Eds pp 297ndash317 ElsevierScience New York NY USA 1994


[152] E Mendez-Alvarez R Soto-Otero I Sanchez-Sellero andM L-R Lamas ldquoIn vitro inhibition of catalase activity bycigarette smoke Relevance for oxidative stressrdquo Journal ofApplied Toxicology vol 18 no 6 pp 443ndash448 1998

[153] H A Nadiger C A Mathew and B Sadasivudu ldquoSerummalanodialdehyde (TBA reactive substance) levels in cigarettesmokersrdquo Atherosclerosis vol 64 no 1 pp 71ndash73 1987

[154] G J Handelman L Packer and C E Cross ldquoDestructionof tocopherols carotenoids and retinol in human plasma bycigarette smokerdquoTheAmerican Journal of Clinical Nutrition vol63 no 4 pp 559ndash565 1996

[155] C E Cross A van der Vliet and J P Eiserich ldquoCigarette smok-ers and oxidant stress a continuing mysteryrdquo The Americanjournal of clinical nutrition vol 67 no 2 pp 184ndash185 1998

[156] D Lapenna S de Gioia A Mezzetti et al ldquoCigarette smokeferritin and lipid peroxidationrdquoAmerican Journal of Respiratoryand Critical Care Medicine vol 151 no 2 pp 431ndash435 1995

[157] A M Preston ldquoCigarette smoking-nutritional implicationsrdquoProgress in Food and Nutrition Science vol 15 no 4 pp 183ndash217 1991

[158] M-J Hosseini P Naserzadeh A Salimi and J PourahmadldquoToxicity of cigarette smoke on isolated lung heart and brainmitochondria induction of oxidative stress and cytochrome creleaserdquo Toxicological and Environmental Chemistry vol 95 no9 pp 1624ndash1637 2013

[159] Y-M Yang and G-T Liu ldquoInjury of mouse brain mitochondriainduced by cigarette smoke extract and effect of vitamin C on itin vitrordquo Biomedical and Environmental Sciences vol 16 no 3pp 256ndash266 2003

[160] K Anbarasi G Vani and C S S Devi ldquoProtective effect ofbacoside A on cigarette smoking-induced brain mitochondrialdysfunction in ratsrdquo Journal of Environmental Pathology Toxi-cology and Oncology vol 24 no 3 pp 225ndash234 2005

[161] A E Speck D Fraga P Soares et al ldquoCigarette smokeinhibits brain mitochondrial adaptations of exercised micerdquoNeurochemical Research vol 36 no 6 pp 1056ndash1061 2011

[162] J R Pauly and T A Slotkin ldquoMaternal tobacco smoking nico-tine replacement and neurobehavioural developmentrdquo ActaPaediatrica vol 97 no 10 pp 1331ndash1337 2008

[163] M Steinlin ldquoCerebellar disorders in childhood cognitive prob-lemsrdquo Cerebellum vol 7 no 4 pp 607ndash610 2008

[164] K Anbarasi G Vani K Balakrishna and C S Devi ldquoEffectof bacoside A on membrane-bound ATPases in the brain ofrats exposed to cigarette smokerdquo Journal of Biochemical andMolecular Toxicology vol 19 no 1 pp 59ndash65 2005

[165] H Rauchova J Ledvinkova M Kalous and Z DrahotaldquoThe effect of lipid peroxidation on the activity of variousmembrane-bound ATPases in rat kidneyrdquo International Journalof Biochemistry and Cell Biology vol 27 no 3 pp 251ndash255 1995

[166] J H Sisson D J Tuma and S I Rennard ldquoAcetaldehyde-mediated cilia dysfunction in bovine bronchial epithelial cellsrdquoThe American Journal of PhysiologymdashLung Cellular and Molec-ular Physiology vol 260 no 2 part 1 pp L29ndashL36 1991

[167] G J Lees ldquoInhibition of sodium-potassium-ATPase a poten-tially ubiquitous mechanism contributing to central nervoussystem neuropathologyrdquo Brain Research Reviews vol 16 no 3pp 283ndash300 1991

[168] AYXiao LWei S Xia S Rothman and S P Yu ldquoIonicmecha-nism of ouabain-induced concurrent apoptosis and necrosis inindividual cultured cortical neuronsrdquo Journal of Neurosciencevol 22 no 4 pp 1350ndash1362 2002

[169] A K Singh ldquoEarly developmental changes in intracellular Ca2+stores in rat brainrdquo Comparative Biochemistry and PhysiologyPart A Molecular and Integrative Physiology vol 123 no 2 pp163ndash172 1999

[170] A Zaidi andM LMichaelis ldquoEffects of reactive oxygen specieson brain synaptic plasmamembraneCa2+-ATPaserdquoFree RadicalBiology and Medicine vol 27 no 7-8 pp 810ndash821 1999

[171] M Brownlee ldquoAdvanced protein glycosylatlon in diabetes andagingrdquo Annual Review of Medicine vol 46 pp 223ndash234 1995

[172] M Brownlee ldquoNegative consequences of glycationrdquoMetabolism vol 49 no 2 pp 9ndash13 2000

[173] P K Janicki J L Horn G Singh W T Franks and J J FranksldquoDiminished brain synaptic plasma membrane Ca2+-ATPaseactivity in rats with streptozocin-induced diabetes associationwith reduced anesthetic requirementsrdquo Life Sciences vol 55 no18 pp PL359ndashPL364 1994

[174] T C Squier and D J Bigelow ldquoProtein oxidation and age-dependent alterations in calcium homeostasisrdquo Frontiers inBioscience vol 5 pp D504ndashD526 2000

[175] N Nedeljkovic G Nikezic A Horvat S Pekovic M Sto-jiljkovic and J V Martinovic ldquoProperties of Mg(2+)-ATPaserat brain synaptic plasma membranesrdquo General Physiology andBiophysics vol 17 pp 3ndash13 1998

[176] A Grochowalska and R Bernat ldquoAdaptacja aktywnosci ATP-az synaptosomow roznych obszarow mozgu w hipoksji mody-fikowanej wpływem adrenergicznym i gabaergicznym (Adap-tation of ATP-ase activity in synaptosomes of various cerebralregions in hypoxia modified by adrenergic and gabaergicinfluences)rdquo Nowiny Lekarskie vol 66 no 4 pp 397ndash412 1997

[177] R Vink T K McIntosh and A I Faden ldquoMagnesium incentral nervous systemrdquo inNeuroscience Year G Adelman EdSupplement 1 to the Encyclopedia of Neuroscience pp 93ndash94Birkhauser Boston Mass USA 1989

[178] H Haga ldquoEffects of dietary magnesium supplementation ondiurnal variations of blood pressure and plasma Na+ K+-ATPase activity in essential hypertensionrdquo Japanese Heart Jour-nal vol 33 no 6 pp 785ndash800 1992

[179] A Rajpurkar Y Jiang C B Dhabuwala J C Dunbar and HLi ldquoCigarette smoking induces apoptosis in rat testisrdquo Journal ofEnvironmental Pathology Toxicology and Oncology vol 21 no3 pp 243ndash248 2002

[180] A J Blaschke J A Weiner and J Chun ldquoProgrammed celldeath is a universal feature of embryonic and postnatal neu-roproliferative regions throughout the central nervous systemrdquoThe Journal of Comparative Neurology vol 396 no 1 pp 39ndash501998

[181] T S Roy J E Andrews F J Seidler and T A Slotkin ldquoNicotineevokes cell death in embryonic rat brain during neurulationrdquoJournal of Pharmacology and Experimental Therapeutics vol287 no 3 pp 1136ndash1144 1998

[182] R Garrido K King-Pospisil K W Son B Hennig and MToborek ldquoNicotine upregulates nerve growth factor expressionand prevents apoptosis of cultured spinal cord neuronsrdquoNeuro-science Research vol 47 no 3 pp 349ndash355 2003

[183] K Anbarasi G Kathirvel G Vani G Jayaraman and C SShyamala Devi ldquoCigarette smoking induces heat shock protein70 kDa expression and apoptosis in rat brain modulation bybacoside Ardquo Neuroscience vol 138 no 4 pp 1127ndash1135 2006

[184] S Dwivedi R Nagarajan K Hanif H H Siddiqui C Nath andR Shukla ldquoStandardized extract of Bacopa monniera attenuatesokadaic acid induced memory dysfunction in rats effect on


Nrf2 pathwayrdquo Evidence-Based Complementary and AlternativeMedicine vol 2013 Article ID 294501 18 pages 2013

[185] R D Shytle A A Silver R J Lukas M B Newman D VSheehan and P R Sanberg ldquoNicotinic acetylcholine receptorsas targets for antidepressantsrdquo Molecular Psychiatry vol 7 no6 pp 525ndash535 2002

[186] M B Newman G W Arendash R D Shytle P C Bickford TTighe and P R Sanberg ldquoNicotinersquos oxidative and antioxidantproperties in CNSrdquo Life Sciences vol 71 no 24 pp 2807ndash28202002

[187] G N Neigh K Karelina E R Glasper et al ldquoAnxiety aftercardiac arrestcardiopulmonary resuscitation exacerbated bystress and prevented by minocyclinerdquo Stroke vol 40 no 11 pp3601ndash3607 2009

[188] P Casolini A Catalani A R Zuena and L AngeluccildquoInhibition of COX-2 reduces the age-dependent increase ofhippocampal inflammatory markers corticosterone secretionand behavioral impairments in the ratrdquo Journal of NeuroscienceResearch vol 68 no 3 pp 337ndash343 2002


(tar) phase of cigarette smoke contains sim1017 free radicalsgand the gas phase contains sim1015 free radicalspuff [12] Theradicals associated with the tar phase are long-lived (hours tomonths) whereas the radicals associated with the gas phasehave a shorter life span (seconds) [12ndash14]

Cigarette smoke that is drawn through the tobacco intoan active smokerrsquos mouth is known as mainstream smoke(MS) and the smoke emitted from the burning ends of acigarette is the sidestream smoke (SS) Mainstream cigarettesmoke comprises 8 of tar and 92 of gaseous compo-nents [12] Environmental tobacco smoke (ETS) results fromthe combination of sidestream smoke (85) and a smallfraction of exhaled mainstream smoke (15) from smokers[13] Importantly the concentration of numerous toxins isdramatically (up to 100-fold) elevated in SS when comparedwith MS and the complex mixture of toxins is attributedrather to a specific component of cigarette smoke to thepotential adverse impact of passive smoke on health [15]

Aside from specific chemical constituents certain physic-ochemical properties of smoke may participate in diseaseprocesses The pH of the smoke affects the site and degreeof nicotine absorption as well as the smokerrsquos depth ofinhalation The oxidation-reduction state of the smoke isimportant because oxidants influence the maturation ofcholesterol-laden plaques in the coronary arteries and otherblood vessels In short cigarette smoke is farmore than a triadof tar nicotine and carbon monoxide [16]

Although SS and MS smoke have qualitatively similarchemical compositions the respective quantities of individ-ual smoke constituents are different [17] The exposure to SSsmoke depends on the distance from the burning cigaretteand conditions of ventilation the higher concentrations ofcertain toxic and carcinogenic chemicals in SS smoke result inmeasurable levels of these chemicals in nonsmokers exposedto ETS [18]

3 Pharmacodynamics of Smoking

Although most of the toxicity of smoking is related to othercomponents of cigarette smoke it is primarily the pharmaco-logic effects of nicotine that produce the addiction to tobaccoAn understanding of how nicotine produces addiction andinfluences smoking behavior provides a necessary basis foroptimal smoking cessation intervention Cigarette smokecontains 10ndash14mg of nicotine [19] of which 1ndash15mg isabsorbed systemically in the lungs through inhalation [20]Nicotine rapidly enters the pulmonary venous circulationreaches the brain within 10ndash20 s and readily diffuses intobrain tissue [21] and binds to nicotine acetylcholine receptors(nAChRs) [22] Acutely cigarette smoking induces posi-tive reinforcing effects including mild euphoria heightenedarousal reduced appetite and reduced stress anxiety andpain [23]

Nicotine in cigarette smoke is alkaline and readilycrosses the blood brain barrier It mediates the stimulationof mesolimbic dopamine system It is also involved inmodulating other classical neurotransmitters in the brainincluding catecholamines serotonin GABA and glutamate

[24] It induces addiction as it shares many properties ofpsychostimulant drugs such as cocaine and amphetamine[25]With repeated exposure to nicotine toxicity gives way totolerance and addiction Overtime tolerance is accompaniedby increased intake of nicotine to attain the same effectsinitially taken which leads to physical dependence com-pelling higher intake lest it induces withdrawal symptomThis leads to reinforcement mechanisms finally leading toaddiction [26] Substances other than nicotine present incigarette smoke are also involved in mediating the harmfuleffects in nervous system Other tobacco smoke constituentsaffect the structural and functional integrity of blood brainbarrier [27 28]

4 Neuropharmacological Effects ofCigarette Smoking

Cigarette smoking is an important environmental agingaccelerator [29] partly because it induces oxidative stress inmultiple organs including the brain and is presented in manydiseases including cognition-related or neurodegeneration-related pathological changes [30]This should be presented todemonstrate a direct linkage between smoking and cognitiveimpairment

The incidence of cerebrovascular diseases (CVDs)increases with cigarette smoking as cessation of smokingdecreases its incidence [31] Smoking is a modifiablerisk factor for stroke [32] primarily due to hypertension[33] Other neurological diseases for which smoking isa risk factor include hypoxia cerebral ischemia cerebralhemorrhage brain infarction subarachnoid hemorrhageand tardive dyskinesia [34] Neuroleptic Parkinsonismresembling Idiopathic Parkinsonrsquos disease is associated withsmoking as higher doses of nicotine exert an inhibitoryeffect [35] Cerebral symptoms like brain atrophy and ataxiaare exaggerated with smoking [36] Reports also suggestthat cigarette smoking is protective in the development ofParkinsonrsquos disease [37] but is an important risk factor inAlzheimerrsquos disease [38] as it accelerates cognitive declineand dementia [39]

5 Role of Passive Smoking

The evidence that active smoking is a risk factor forcardiovascular disease (CAD) and the leading cause ofpreventable death is overwhelming However exposure topassive cigarette smoke also exerts detrimental effects onvascular homoeostasis [40] Importantlymost of these effectsappear to be characterized by a rapid onset For example therelatively low doses of toxins inhaled by passive smoking aresufficient to elicit acute endothelial dysfunction and theseeffects may be related at least in part to the inactivation ofnitric oxide (NO) Moreover passive smoking may directlyimpair the viability of endothelial cells and reduce thenumber and functional activity of circulating endothelialprogenitor cells In addition platelets of nonsmokers appearto be susceptible to proaggregatory changes with everypassive smoke exposure Overall passive smoke induces










7 Methods


















(a)times10)(H amp E

(b)

times40)(H amp E


times40)(H amp E


120572

120573

120575

120579


Control CS BA


Control


CS BA CS + BA

CS + BA







times100

(a)

times100

(b)

times100

(c)times100

(d)


NS

0

5

10

15

20

25

30

35

Control CS BA

(nm

oles

g ti

ssue

)

CS + BA

F valueACh = 14948

lowast

lowast







NS

NS

NSNS

0

10

20

30

40

50

60

Control CS BA

(ng

g tis

sue)


Norepinephrine

Epinephrine

CS + BA

F value

lowast

lowast

lowastlowast

lowast

lowast

lowast lowast




NS

NS

NSNS

0

10

20

30

40

50

60

Control CS BA

(ng

g tis

sue)


lowast

lowastlowast

lowast

lowast

lowast

lowast

lowast

CS + BA




NS

0100200300400500600700800900

Control CS BA

min

mg

prot

ein)

(120583

mol

es o

f thi

ocho

line l

iber

ated

CS + BA

F valueAChE = 1015

lowast

lowast


NS

0010203040506070809

Control CS BA

liber

ated

min

mg

prot

ein)

(120583m

oles

of b

enzla

dehy

de

CS + BA

F valueMAO = 1015

lowast

lowast







Bacoside A(BA)


















Bacoside A(BA)





nd not detected










9 Conclusion






Acknowledgment


References














































4120573




































































































































































7 Methods


















(a)times10)(H amp E

(b)

times40)(H amp E


times40)(H amp E


120572

120573

120575

120579


Control CS BA


Control


CS BA CS + BA

CS + BA







times100

(a)

times100

(b)

times100

(c)times100

(d)


NS

0

5

10

15

20

25

30

35

Control CS BA

(nm

oles

g ti

ssue

)

CS + BA

F valueACh = 14948

lowast

lowast







NS

NS

NSNS

0

10

20

30

40

50

60

Control CS BA

(ng

g tis

sue)


Norepinephrine

Epinephrine

CS + BA

F value

lowast

lowast

lowastlowast

lowast

lowast

lowast lowast




NS

NS

NSNS

0

10

20

30

40

50

60

Control CS BA

(ng

g tis

sue)


lowast

lowastlowast

lowast

lowast

lowast

lowast

lowast

CS + BA




NS

0100200300400500600700800900

Control CS BA

min

mg

prot

ein)

(120583

mol

es o

f thi

ocho

line l

iber

ated

CS + BA

F valueAChE = 1015

lowast

lowast


NS

0010203040506070809

Control CS BA

liber

ated

min

mg

prot

ein)

(120583m

oles

of b

enzla

dehy

de

CS + BA

F valueMAO = 1015

lowast

lowast







Bacoside A(BA)


















Bacoside A(BA)





nd not detected










9 Conclusion






Acknowledgment


References














































4120573








































































































































































(a)times10)(H amp E

(b)

times40)(H amp E


times40)(H amp E


120572

120573

120575

120579


Control CS BA


Control


CS BA CS + BA

CS + BA







times100

(a)

times100

(b)

times100

(c)times100

(d)


NS

0

5

10

15

20

25

30

35

Control CS BA

(nm

oles

g ti

ssue

)

CS + BA

F valueACh = 14948

lowast

lowast







NS

NS

NSNS

0

10

20

30

40

50

60

Control CS BA

(ng

g tis

sue)


Norepinephrine

Epinephrine

CS + BA

F value

lowast

lowast

lowastlowast

lowast

lowast

lowast lowast




NS

NS

NSNS

0

10

20

30

40

50

60

Control CS BA

(ng

g tis

sue)


lowast

lowastlowast

lowast

lowast

lowast

lowast

lowast

CS + BA




NS

0100200300400500600700800900

Control CS BA

min

mg

prot

ein)

(120583

mol

es o

f thi

ocho

line l

iber

ated

CS + BA

F valueAChE = 1015

lowast

lowast


NS

0010203040506070809

Control CS BA

liber

ated

min

mg

prot

ein)

(120583m

oles

of b

enzla

dehy

de

CS + BA

F valueMAO = 1015

lowast

lowast







Bacoside A(BA)


















Bacoside A(BA)





nd not detected










9 Conclusion






Acknowledgment


References














































4120573




























































































































































times100

(a)

times100

(b)

times100

(c)times100

(d)


NS

0

5

10

15

20

25

30

35

Control CS BA

(nm

oles

g ti

ssue

)

CS + BA

F valueACh = 14948

lowast

lowast







NS

NS

NSNS

0

10

20

30

40

50

60

Control CS BA

(ng

g tis

sue)


Norepinephrine

Epinephrine

CS + BA

F value

lowast

lowast

lowastlowast

lowast

lowast

lowast lowast




NS

NS

NSNS

0

10

20

30

40

50

60

Control CS BA

(ng

g tis

sue)


lowast

lowastlowast

lowast

lowast

lowast

lowast

lowast

CS + BA




NS

0100200300400500600700800900

Control CS BA

min

mg

prot

ein)

(120583

mol

es o

f thi

ocho

line l

iber

ated

CS + BA

F valueAChE = 1015

lowast

lowast


NS

0010203040506070809

Control CS BA

liber

ated

min

mg

prot

ein)

(120583m

oles

of b

enzla

dehy

de

CS + BA

F valueMAO = 1015

lowast

lowast







Bacoside A(BA)


















Bacoside A(BA)





nd not detected










9 Conclusion






Acknowledgment


References














































4120573




























































































































































NS

NS

NSNS

0

10

20

30

40

50

60

Control CS BA

(ng

g tis

sue)


Norepinephrine

Epinephrine

CS + BA

F value

lowast

lowast

lowastlowast

lowast

lowast

lowast lowast




NS

NS

NSNS

0

10

20

30

40

50

60

Control CS BA

(ng

g tis

sue)


lowast

lowastlowast

lowast

lowast

lowast

lowast

lowast

CS + BA




NS

0100200300400500600700800900

Control CS BA

min

mg

prot

ein)

(120583

mol

es o

f thi

ocho

line l

iber

ated

CS + BA

F valueAChE = 1015

lowast

lowast


NS

0010203040506070809

Control CS BA

liber

ated

min

mg

prot

ein)

(120583m

oles

of b

enzla

dehy

de

CS + BA

F valueMAO = 1015

lowast

lowast







Bacoside A(BA)


















Bacoside A(BA)





nd not detected










9 Conclusion






Acknowledgment


References














































4120573






























































































































































Bacoside A(BA)


















Bacoside A(BA)





nd not detected










9 Conclusion






Acknowledgment


References














































4120573






























































































































































Bacoside A(BA)





nd not detected










9 Conclusion






Acknowledgment


References














































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Acknowledgment


References














































4120573
















































































































































































4120573



























































































































































bacoside a: role in cigarette smoking induced changes in brain · 2018. 11. 27. · bacoside a:...

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