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[6]-Gingerol isolated from ginger attenuatessodium arsenite induced oxidative stress and
play s a corrective role in improving insulin
signaling in mice
ARTICLE in TOXICOLOG Y LETTERS · JANUARY 2012
Impact Factor: 3.26 · DOI: 10.1016/j.toxlet.2012.01.002 · Source: PubMed
CITATIONS
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6 AUTHORS, INCLUDING:
Debrup Chakraborty
Michigan State University
12 PUBLICATIONS 191 CITATIONS
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Avinaba Mukherjee
Jadavpur University
21 PUBLICATIONS 203 CITATIONS
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Samrat Ghosh
University of Kalyani
15 PUBLICATIONS 211 CITATIONS
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Anisur Rahman Khuda-Bukhsh
University of Kalyani
315 PUBLICATIONS 2,636 CITATIONS
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Toxicology Letters 210 (2012) 34–43
Contents lists available at SciVerse ScienceDirect
Toxicology Letters
journal homepage: www.elsevier .com/ locate / toxlet
[6]-Gingerol isolated from ginger attenuates sodium arsenite induced oxidativestress and plays a corrective role in improving insulin signaling in mice
Debrup Chakraborty, Avinaba Mukherjee, Sourav Sikdar, Avijit Paul, Samrat Ghosh,Anisur Rahman Khuda-Bukhsh∗
Cytogenetics andMolecular Biology Laboratory, Department of Zoology, University of Kalyani, Kalyani 741235, West Bengal, India
a r t i c l e i n f o
Article history:
Received 28 November 2011
Received in revised form
30 December 2011
Accepted 2 January 2012
Available online xxx
Keywords:
Sodium arsenite
[6]-Gingerol
Oxidative stress
Hyperglycemia
GLUT4
Insulin signaling
a b s t r a c t
Arsenic toxicity induces type 2 diabetes via stress mediated pathway. In this study, we attempt to reveal
how sodium arsenite (iAs) could induce stress mediated impaired insulin signaling in mice and if an
isolated active fraction of ginger, [6]-gingerol could attenuate the iAs intoxicated hyperglycemic condi-
tion of mice and bring about improvement in their impaired insulin signaling. [6]-Gingerol treatment
reduced elevated blood glucose level and oxidative stress by enhancing activity of super oxide dismutase
(SOD), catalase, glutathione peroxidase (GPx) and GSH. [6]-Gingerol also helped in increasing plasma
insulin level, brought down after iAs exposure. iAs treatment to primary cell culture of -cells andhepatocytes in vitro produced cyto-degenerative effect and accumulated reactive oxygen species (ROS)
in pancreatic -cells and hepatocytes of mice. [6]-Gingerol appeared to inhibit/intervene iAs inducedcyto-degeneration of pancreatic -cells and hepatocytes, helped in scavenging the free radicals. Theover-expression of TNF and IL6 in iAs intoxicated mice was down-regulated by [6]-gingerol treatment.iAs intoxication reduced expression levels of GLUT4, IRS-1, IRS-2, PI3K, AKT, PPAR signaling molecules;[6]-gingerol mediated its action through enhancing the expressions of these signaling molecules, both
at protein and mRNA levels. Thus, our results suggest that [6]-gingerol possesses an anti-hyperglycemicproperty and can improve impaired insulin signaling in arsenic intoxicated mice.
© 2012 Elsevier Ireland Ltd. All rights reserved.
1. Introduction
Arsenic is a naturally occurring heavy metal that is present in
food, soil and water. It is released in the environment from both
natural and man-made sources (Tchounwou et al., 1999). Inorganic
arsenic and theirmetabolites (both As+3 andAs+5 forms) are known
to exert their toxic effects by a variety of mechanisms which may
lead to some serious health problems. Epidemiological data have
shown that chronic exposure of inorganic arsenical compounds to
humans are associatedwith liverinjury,peripheral neuropathy andanincreasedincidence of cancer of thelung, skin,and liver (Leonard
andLauwerys, 1980). In EastAsia alone, including Bangladesh, West
Bengal, India, Vietnam, Thailand and China, more than 30 million
people are chronically exposed to arsenic (Tseng et al., 1968). This
arsenic induced toxicity arises and sustains by generating stress
response through reactive oxygen species formation and antioxi-
dant depletion ( Jomova et al., 2011). According to a recent study,
sodium arsenite (iAs)is found to be associatedwith increasedblood
glucose level in experimental rats (Yousef et al., 2008).
∗ Corresponding author. Tel.: +91 33 25828750x315.
E-mail addresses:khudabukhsh [email protected], prof [email protected]
(A.R. Khuda-Bukhsh).
Hydroarsenicism is a major public health problem since mil-
lions of people worldwide are exposed to arsenic by drinking of
contaminated water ( Jones, 2007). Studies on mouse bone marrow
cells have predicted an increased level of chromosomal abnor-
mality and micronucleus formation after treatment with arsenic
(Banerjee et al., 2007) andtherebyhaveconfirmed itscytotoxic and
cytodegenerative effects. One of the plausible modes of action of
arsenic toxicity is by oxidative stress since it can stimulate produc-
tion of reactive oxygen species (ROS), resulting from an imbalance
between antioxidants and oxidants during arsenic metabolism(Goering et al., 1999; Sun et al., 2006).
On the other hand arsenic has been recently proposed as
an additional risk factor for diabetes (Silbergeld et al., 2008;
Longnecker and Daniels, 2001). According to recent surveys it is
found that the occurrence of diabetes is significantly higher in
arsenic-endemic villages in Taiwan and India than in the general
population (Zimmet, 1982; Wang et al., 1997; Belon et al., 2006).
The prevalence of diabetesmellituswas 2-fold higher in these areas
than in Taipei City and the Taiwan area in general.
From in vitro studies, the impairment of insulin secretion
(Diaz-Villaseñor et al., 2006) and the induction of oxidative stress
(Izquierdo-Vega et al., 2006) have been postulated for arsenic-
induced type 2 diabetes. Induction of stress via generation of free
oxygen radicals and antioxidant depletionled into this process and
0378-4274/$ – seefront matter © 2012 Elsevier Ireland Ltd. All rights reserved.
doi:10.1016/j.toxlet.2012.01.002
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36 D. Chakraborty et al./ Toxicology Letters 210 (2012) 34–43
Fig. 1. Mass spectra and structure of [6]-gingerol.
2.8. Animal treatment
After selectionof theoptimum dose of iAs(3 mg/kg), animals were randomized
and 36 mice were divided into six groups, consisting of six mice in each group and
they were treated for 15 weeks as follows:
Group 1: Normal control: animals received only water as vehicle.
Group 2: Arsenic control (iAs): animals received iAs at 3 mg/kg body weight oncedaily for12 weeks, orally.
Group 3: [6]-Gingeroltreated group (iAs+ [6]-gingerol): received iAs for12 weeks
followed by [6]-gingeroladministrationat a dose of 50mg/kgbody weightin alco-
hol once daily for next 3 weeks.
Group 4: [6]-Gingeroltreated group (iAs+ [6]-gingerol): received iAs for12 weeks
followed by [6]-gingeroladministrationat a dose of 75mg/kgbody weightin alco-
hol once daily for next 3 weeks.
Group 5: Alcohol treated group (iAs+ alcohol): received iAs for12 weeks followed
by alcohol administration (equal quantity as administeredin groups 3 and4) once
dailyfor next 3 weeks.
Group 6: [6]-Gingerol alone treated group: normal mice were treated with [6]-
gingerol (orally, 75mg/kg body weight, once daily)for 3 weeks to see whether the
drug alone has anyadverse effect on mice.
The animals were humanely sacrificed under light ether anesthesia and livers
were collected.
We fedthe drug orallyto all themice of differentexperimental groups throughgavage. No significant changes in ALT, AST activity were found between iAs control
and alcohol (drug vehicle) treated groups. There were also no significant differ-
ences of ALT and AST activities between normal control and [6]-gingerol alone
treated groups. Therefore, we excluded groups 5 and 6 from more in-depth stud-
ies. Mice showing blood glucose levels of more than 180mg/dl were considered as
hyperglycemic animals and included in our experiments.
2.9. Collection of blood and tissue samples
At the end of the experimental period, we kept t he animals on f as t f or 12h
prior tobeingsacrificed fordetermining glucoselevel intheirfasting blood.We also
collected the pancreas and liver tissues from each experimental animal and stored
them at −80 ◦C for further analysis.
2.10. Preparation of liver tissue homogenates
We collected the liver tissues from experimental mice, homogenized them in
lysisbuffer usingglass homogenizer and centrifugedat 12,000× g for30 min at 4 ◦C.
We collectedthe supernatant aftercentrifugation and usedit forfurtherexperiment
andestimated thetotal proteinaccordingto themethodof Bradfordusing crystalline
bovineserum albumin(BSA) as standard (Bradford, 1976).
2.11. Determination of pancreatic and hepatic arsenic contents
The arsenic contents of liver tissues of all experimental animals were analyzed
according to themethod described by Khuda-Bukhshet al. (2005), using an atomic
absorption spectrophotometer (AAS).
2.12. Determination of in vivo antioxidant capacity
We determined antioxidant capacity of [6]-gingerol on hepatic tissues of all
experimental animals by FRAP (ferric reducing antioxidantpotential) assay( Benzie
and Strain, 1999). We took the absorbance of the sample against reagent blank
(1.5ml FRAP reagent +50 l distilled water)at 593nm (Manna et al., 2009).
2.13. Biochemical analysis of blood and activity of antioxidant markers in liver
We estimated thebloodglucose level using a glucose estimation kit(Accu-chek
active), Rochediagnostics, Mannheim,Germany.We used livertissue homogenates
for various enzymatic assays. We undertookspectrophotometricanalysisof activity
of catalase (CAT) (Aebi, 1984), super oxide dismutase (SOD) (Fridovich, 1989), glu-
tathione peroxidase(GPx),level of totalglutathione (GSH) (Ellman, 1959) according
to the standard protocols.
2.14. Oral glucose tolerance test (OGTT)
Anoral glucose tolerancetest (OGTT) wasperformed onthe lastday of treatment
after overnight fasting. Blood wascollected from the tail vein of mice at time 0, 60,
90 and120 min after anoral glucose load of 3.0g/kgof body weight. Only water was
provided inside the cages during thecourse of experiment.
2.15. ELISA for activity measurement of different antibodies
We assayed the activity of plasma insulin and hepatic GLUT4 according to
the manufacturer’s protocol (Santa Cruz Biotechnology, Inc., USA) and quantified
t he m using an ELIS A r eader ( The rm o Scie nt ifi c, USA). We used pNPP (para-
nitrophenylphosphate) asa colordevelopingagent andmeasuredthe colorintensity
in 405nm wavelength.
2.16. Immunoblot analysis
We usedthe livertissue homogenates for immunoblotanalysis. Forthis we took
50mg oftissuesamplesin2mllysisbufferforproteinextraction.We undertookSDS-
PAGE (12.5%) electrophoresis of equal amounts of lysate protein and transferred
them to polyvinyl difluoride (PVDF) membrane. After blocking with 3% BSA, we
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D. Chakraborty et al./ Toxicology Letters 210 (2012) 34–43 37
Fig. 2. Effect of [6]-gingerol on the viability of iAs intoxicated hepatocytes and
pancreatic -cells. While the-cells were exposed to 10M iAs and differentcon-
centrations of [6]-gingerol for 72h, the hepatocytes were exposed for 8 h. The cell
viability was thendetected by MTT assay. Eachpoint expressedas mean±SD (N =6).
Significance, * p< 0.05 vs.normal control group. Significance,# p
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38 D. Chakraborty et al./ Toxicology Letters 210 (2012) 34–43
Fig. 3. Dose dependent inhibitory effect of [6]-gingerol on iAs induced free radical accumulation. 1 h after incubation with iAs (10M) cells were incubated with 50 and
75g/ml doses of [6]-gingerol. -Cells and hepatocytes incubated for next 71h and 7h, respectively with drug. A–D represents fluorescence microscopic observations
and E–H represents flowcytometric analysis of ROS accumulation in -cells. On the other hand, I–L represents fluorescence microscopic observations and M–P represents
flowcytometric analysis of ROSaccumulation in hepatocytes.A, E, I, M – normalcontrol cells,B, F, J, N – 10M iAsintoxicatedcells, C, G, K, O – iAsintoxicatedcells, incubated
with 50g/ml [6]-gingerol, D, H, L, P – iAs intoxicated cells, incubated with 75g/ml [6]-gingerol. The intra-cellular ROS was detected by DCFHDA method. Q–T represents
flowcytometric analysis of intracellular GLUT4 content in hepatocytes. Q – normal control cells, R – 10M iAs intoxicated cells, S – iAs intoxicated cells, incubated with
50g/ml [6]-gingerol, T – iAs intoxicated cells, incubated with 75g/ml [6]-gingerol.
Table 1
Effect of different concentrations of iAs on mice.
Cont 2 mg/kg 3 mg/kg 4 mg/kg 5 mg/kg
ALT (m/mg protein) 11.06 ± 3.10 16.6 ± 3.109 23.24 ± 0.560* 25.04 ± 0.66* 27.93 ± 1.17*
AST (m/mg protein) 5.33 ± 0.203 6.55 ± 1.66 14.007 ± 0.509* 13.86 ± 0.441* 17.32 ± 2.68*
Blood glucose (mg/dl) 92.5 ± 2.12 132 ± 5.65* 189.5 ± 7.77* 186 ± 8.48* 186 ± 9.89*
Data are expressedas mean±SD (N =6).* p< 0.05 vs. normal control group.
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D. Chakraborty et al./ Toxicology Letters 210 (2012) 34–43 39
Table 2
Dose dependent Influence of [6]-gingerol on iAs intoxicated hyperglycemic mice.
Cont iAs 25 mg/kg 50 mg/kg 75 mg/kg
ALT (m/mg protein) 11.06 ± 3.109 23.24 ± 0.56* 21.87 ± 1.37 15.55 ± 1.35# 13.51 ± 0.45#
AST (m/mg protein) 5.33 ± 0.203 14.007 ± 0.51* 12.78 ± 1.08 8.68 ± 0.71# 8.29 ± 2.2#
Blood g lucose (mg/dl) 92.5 ± 2.12 189.5 ± 7.77* 182 ± 8.48 122.5 ± 3.53# 110 ± 2.82#
Data are expressed as mean±SD (N =6).* p< 0.05 vs. normal control group.
# Significance, p< 0.05 vs. iAs intoxicated group.
Table 3
Role of [6]-gingerolon arsenic deposition in pancreas and liver tissues of experimental animals expressed in nmol/g unit.
Cont iAs 50 mg/kg 75 mg/kg
Pancreas 13.87 ± 4.2 43.015 ± 6.86* 26.435 ± 3.41 23.669 ± 3.74#
Liver 14.92 ± 6.71 49.27 ± 7.73* 27.51 ± 3.11# 25.13 ± 4.14#
Data are expressed as mean±SD (N =6).* p< 0.05 vs. normal control group.
# Significance, p< 0.05 vs. iAs intoxicated diabetic group.
Table 4
Role of [6]-gingerolon different anti-oxidant biomarkers of iAs intoxicated mice liver.
Cont iAs 50 mg/kg 75 mg/kg
GHb (%) 4.02 ± 0.28 10 ± 0.19* 7.05 ± 0.27# 5.32 ± 0.97#
SOD (m/mg protein) 38.68 ± 1.26 28.32 ± 1.73* 32.19 ± 1.29 35.35 ± 0.35#
Catalase (m/mg protein) 72.22 ± 4.08 37.2032 ± 2.97* 52.16 ± 3.54# 58.547 ± 0.95#
GSH (m/mg protein) 37.62 ± 2.65 21.92 ± 1.21* 28.53 ± 1.4 32.29 ± 3.65#
GPx (m/mg protein) 64.5 ± 5.82 48.36 ± 1.44* 54.16 ± 1.59 58.18 ± 2.36#
FRAP (%) 95.83 ± 5.89 54.16 ± 5.89* 67.91 ± 1.76 79.16 ± 5.89#
Data are expressed as mean±SD (N =6).* p< 0.05 vs. normal control group.
# Significance, p< 0.05 vs. iAs intoxicated diabetic group.
3.9. Dose dependent study of [6]-gingerol by FRAP assay
Dose dependent effect of [6]-gingerol against iAs toxicity has
been represented in Table 4. The intracellular ferric reducing
antioxidant potential decliningafter iAsintoxicationin animalswas
found to increase significantly with[6]-gingerol treatment at doses
of 50mg/kg and 75mg/kg bw, respectively.
3.10. Effect on plasma insulin and hepatic GLUT4 content
Under iAs intoxicated hyperglycemic condition, decreased lev-
els of plasma insulin andhepaticGLUT4were found. Administration
Fig.4. Impactof [6]-gingerol(50 and 75 mg/kg)treatment on oralglucose tolerance
in iAs mice. Data were expressed as mean±SD (N =6), Ap< 0.05 cont vs. iAs intox-
icated group and ap< 0.05 iAs vs. drug groups (A/a used to denote comperisons in
0 min interval groups, B/b used to denote comperisons in 60min interval groups,
C/c used to denotecomperisonsin 90min interval groups, D/dused to denotecom-
perisons in 120min interval groups).
of [6]-gingerol at both the doses increased the plasma insulin and
hepatic GLUT4 concentrations significantly (Fig. 5).
3.11. Immunoblot analysis
Administration of the drug down regulated the expression of
TNFwhichwas earlier found to increase in iAsintoxicated liver.Inaddition,we observedthat iAsintoxicationreducedthe expressions
of proteins related to insulin signaling, namely, Insulin, IRS1, IRS2,
AKT, PI3K, GLUT4 and PPAR (Figs. 6–7). However, expressions of these proteins were significantly up regulated after treatment with
[6]-gingerol.
Fig.5. Effectof [6]-gingerolon plasmainsulinand hepatic GLUT4 levelin iAsintoxi-
cateddiabeticmice.Data wereexpressed in histogramas mean±SD (N =6).* p
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40 D. Chakraborty et al./ Toxicology Letters 210 (2012) 34–43
Fig. 6. Immunoblot analysis of insulin, IRS1, TNF, IRS2, GLUT4, PI3K, AKT, PPAR
andGAPDH. GAPDH is used as an equal loading housekeepinggene.Lane 1. Normal
mice; Lane 2. iAs-intoxicated mice, Lane 3. iAs-intoxicated+ [6]-gingerol 50mg/kg
bw, Lane 4. iAs-intoxicated+ [6]-gingerol75 mg/kg bw. Signs indicate that a partic-
ular treatmenthas not(−) been or has been (+)given,respectively.
3.12. RT-PCR analysis
Results of RT-PCR confirmed that there was a significant dif-
ference in the expressions of mRNA between the control and
the iAs-intoxicated groups and between the iAs and drug-treated
groups (Figs. 8–9). Results of RT-PCR analysis also supportedthe results obtained from the western blot analysis. The primer
sequences of amplified genes are given in Table 5.
4. Discussion
Our experimental findings showed that iAs intoxication can
significantly reduce the pancreatic -cells and hepatocyte cell via-bility. But iAs intoxicated cells incubated with [6]-gingerol had the
Table 5
Primer sequences.
Primer name Primer s equences
TNF Fwd 5-GCACAGAAAGCATGATCCGC-3
Rev 5-CTTGGTGGTTTGCTACGACG-3
IL6 Fwd 5-AACGATGATGCACTTGCAGA-3
Rev 5-GAGCATTGGAAATTGGGGTA-3
IRS1 Fwd 5-AGAGTGGTGGAGTTGAGTTG-3
Rev 5-GGTGTAACAGAAGCAGAAGC-3
IRS2 Fwd 5-GGATAATGGTGACTATACCGAGA-3
Rev 5-CTCACATCGATGGCGATATAGTT-3
PI3K Fwd 5-TTAAACGCGAAGGCAACGA-3
Rev 5-CAGTCTCCTCCTGCTGCTGAT-3
AKT Fwd 5-CCTGGACTACCTGCACTCTCGGAA-3
Rev 5-TTGCTTTCAGGGCTGCTCAAGAAGG-3
GLUT4 Fwd 5-AAGATGGCCACGGAGAGAG-3
Rev 5-GTGGGTTGTGGCAGTGAGTC-3
GAPDH Fwd 5-CCATGTTCGTCATGGGTGTGAACCA-3
Rev 5-GCCAGTAGAGGCAGGGATGATGTTC-3
PPAR Fwd 5-GCGGAGATCTCCAGTGATATC-3
Rev 5-TCAGCGACTGGGACTTTTCT-3
potential to partially overcome the iAs induced toxicity and couldreduce the cell death, in vitro. It is reported that arsenic produces
hepatocyte dysfunction including the reduced cell viability (Das
et al., 2010). On the other hand, ROS also plays an important role
in insulin resistance which is a highly prevalent condition impli-
cated in the development of diabetes. We assessed the changes
in free radical accumulation in -cells and hepatocytes and foundan increased level of ROS accumulation in cells intoxicated with
only iAs. On the other hand,treatment with [6]-gingerol along with
iAs intoxication showed lesser amount of ROS accumulation and
significantly increased cell viability in both pancreatic -cells andhepatocytes.
Recently researchers publishedthat arsenic has a profoundtoxic
and cytodegenerative effects on pancreas and liver (Pi et al., 2002;
Santraetal.,2000). Oneof themajor reasons behind arsenic inducedhepatotoxicity is the depletion of antioxidant defense mechanism
which is related to the reductionof anti-oxidative enzymes’ activity
includingSOD,CAT,GPxandGSH(Das etal., 2010). Arsenic in differ-
ent forms also increases the levels of free hydroxyl radicals, which
playan importantrole in the development of genotoxicity(Liuet al.,
2001). Therefore, any agent which by itself is non-toxic, but has
the ability to reduce oxidative stress will be desirable to antago-
nize ROS generation and also thereby genotoxicity. Our findings
Fig. 7. Relative band intensities of immunoblots.The relative intensities of bands were determined using ‘ImageJ’ software.The results shown in histograms arethe average
±SD (N = 6). Significance * p< 0.05 iAs-intoxicated vs. normal control group; significance # p< 0.05 drug-fed vs. iAs-intoxicated group.
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Fig. 8. Reverse transcription polymerase chain reaction analysis of TNF, IL6,
IRS1, I RS2, PI 3K, AKT, PP AR and GA PDH. Lane 1. Normal mic e; La ne 2. iAs-
intoxicated mice, Lane 3. iAs-intoxicated+ [6]-gingerol 50mg/kg bw, Lane 4.
iAs-intoxicated+ [6]-gingerol 75mg/kg bw. Signs indicate that a particular treat-
menthas not (−) been or has been (+)given,respectively.
indicate that the administration of [6]-gingerol reduced the oxida-
tive stress as revealed from the increased activity of antioxidant
biomarkers like CAT, SOD, GPx and GSH. On the other hand, it also
had an anti-hyperglycemic effect as the increased blood glucose
level due to iAs induction was found to decrease up to the normal
level after treatment with [6]-gingerol. In addition, mice treated
with [6]-gingerol also showed significant improvement in oral glu-
cose tolerance. GHbis an importantindex of diabetes management
and we observed a significant fall in GHb after administration of
[6]-gingerol in iAs intoxicated hyperglycemic mice. The in vivo
antioxidant potential of [6]-gingerol in liver tissue was determined
by FRAP assay, which indicates the anti-oxidative potentials of
[6]-gingerol at two different concentrations. The analysis of AAS
data on arsenic content of liver and pancreas of arsenic intoxicated
mice before and after administration of [6]-gingerol confirmed the
ability of this drug to removethe accumulatedarsenic content from
these organs.
According to several authors (Cemek et al., 2008; Singh et al.,
2009), a drug which has the combined anti-oxidative and anti-hyperglycemic properties can make the drug very effective as
an anti-diabetic drug. Pancreatic -cell dysfunction due to iAstoxicity is the main reason behind impaired insulin secretion
which plays an important role in the onset of type 2 diabetes. In
in vivo set of experiment, treatment with both the doses (50mg/kg
and 75mg/kg bw) of [6]-gingerol was found to be associated
with increased plasma insulin concentration in iAs intoxicated
mice.
iAs has been shown to potentially alter some gene expressions
related to glucose homeostasis in body. Uptakeof glucose molecule
inside the cell depends on the translocation of glucose transporter
4 (GLUT4), which plays a very important role in insulin signaling,
by the action of insulin molecule to the plasma membrane. Hence,
we targeted the intracellular GLUT4 content in isolated hepato-cytes (control cells:62.2%) and found that 10M of iAsintoxicationlowers the level of GLUT4 content (31.3%), which was increased
(up to 38.9% and 43%) after incubation with [6]-gingerol. We also
assessed the mRNA and protein level expressions of GLUT4 in iAs
intoxicated mice, treated or un-treated with [6]-gingerol. Auto-
phosphorylation of insulin receptor substrates and activation of
PI3K/AKT signaling pathwayare required for the activation of down
steam signal cascade leading to glucose uptake and metabolism
in cell (Vijayakumar et al., 2005). Therefore we investigated the
expressions of IRS1, IRS2, p85 of PI3K and AKT in liver tissue of
experimental mice.Our results revealed that[6]-gingerol enhanced
the activity and expressions of these genes which were initially
down regulated in iAs intoxicated hyperglycemic mice. TNF- andIL6 levels are closely associated with both hyper-insulinemia and
insulin resistance(Rondinone,2006). Increasedlevels of TNF- andIL6 are also associated with iAs exposure. Therefore, we examined
the changes of expressions of these two genes in iAs intoxicated
mice, and after treatment with [6]-gingerol, reduced expression
levels of thesaid genes were found. This would revealthat thedrug
exerted its effect in combating impaired insulin responsiveness
through regulation of these genes. iAs intoxication can also down
regulate thetranscription factor PPAR whichisverycloselyrelatedto the activity of TNF- and IL6 (Gurnell, 2003). Our investigation
Fig. 9. Relative fluorescence intensities of PCR bands. The relative intensities of bands were determined using ‘Image J’ software. The results shown in histograms are the
average ±SD (N = 6). Significance * p< 0.05 iAs-intoxicated vs. normal control group; significance # p< 0.05 drug-fed vs. iAs-intoxicated group.
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