antiobesity effect of codonopsis lanceolata in high-calorie/high-fat-diet-induced obese rats

Hindawi Publishing CorporationEvidence-Based Complementary and Alternative MedicineVolume 2013 Article ID 210297 9 pageshttpdxdoiorg1011552013210297

Research ArticleAntiobesity Effect of Codonopsis lanceolata inHigh-CalorieHigh-Fat-Diet-Induced Obese Rats

Hye-Kyung Choi1 Eun-Kyung Won1 Young Pyo Jang2 and Se-Young Choung1

1 Department of Preventive Pharmacy and Toxicology College of Pharmacy Kyung Hee University Seoul 130-701 Republic of Korea2Division of Pharmacognosy College of Pharmacy Kyung Hee University Seoul 130-701 Republic of Korea

Correspondence should be addressed to Se-Young Choung sychoungkhuackr

Received 25 November 2012 Revised 2 April 2013 Accepted 17 April 2013

Academic Editor Srinivas Nammi

Copyright copy 2013 Hye-Kyung Choi et al This is an open access article distributed under the Creative Commons AttributionLicense which permits unrestricted use distribution and reproduction in any medium provided the original work is properlycited

The antiobesity effects of Codonopsis lanceolata (CL) were evaluated in a high-caloriehigh-fat-diet (HFD-) induced obesity ratmodel and 3T3-L1 cellsThe Sprague-Dawleymale rats were fed a normal diet (ND) or a HFD for a period of 12 weeksThe rats weresubdivided into groups ND ND + wild Codonopsis lanceolata (wCL) (900mgkgday po) ND + cultivated Codonopsis lanceolata(cCL) (900mgkgday po) HFD HFD + wCL (100 300 or 900mgkgday po) HFD + cCL (100 300 or 900mgkgday po)and HFD + sibutramine The body weight gains of the administered HFD + CL (wCL or CCL) were lower than those of the ratsfed with only the HFD group Moreover the weight of adipose pads and the serum levels of triglycerides total cholesterol andlow density lipoprotein cholesterol in the group administered HDL + CL were significantly lower than in the HFD group Theinhibitory effect of lipid accumulation in 3T3-L1 cells was measured by Oil Red O staining and reverse transcription-polymerasechain reaction (RT-PCR) Treatment of 3T3-L1 cells with wCL inhibited lipid accumulation and expression of CEBP120572 and PPAR120574These results suggest that CL has a great potential as a functional food with anti-obesity effects and as a therapeutic alternative inthe treatment of obesity

1 Introduction

As obesity has increased in the population multiple methodsof dieting to achieve weight loss have been introducedAmong these diets controlling calorie intake and increasingcalorie consumption expenditure through a proper exercisehave been considered as the ideal However a large numberof people depend on fasting and the consumption of dietfood and medicine to control their weight This tendency hasaffected food consumption trends and has led to an increasein the consumption of functional foods for antiobesity lead-ing to a strong demand for the development of new sourcesof functional foods and related studies on their efficacy

In recent years there have been many efforts to findpromising functional foods that can be used on a dailybasis to improve physical constitution A good example ofthis is Clerodendron glandulosum Coleb This herb foundin north eastern states of India [1] is a folklore medicineconsumed by urban and tribal populace of Manipur againstobesity diabetes andhypertension [2 3] Recently it has been

reported that aqueous extracts from Clerodendron glandulo-sumColeb can regulate high-fat-diet induced hyperlipidemiain rats [3] improve fructose induced insulin resistance andhypertension [4] and prevent high-fat-diet-induced hepaticsteatosis [5]

As part of these recent endeavors this study was per-formed to investigate the influence of the water extracts ofwild and cultivated Codonopsis lanceolata on the serum andbody fat levels of rats fed a high-fat-diet Codonopsis lanceo-lata (CL) is a perennial plant in the family of Campanulaceaeoften called Baishen or Shashen in China CL can be classifiedasBei Sha Shen which shows a red surface andhas a thick rootandmany rootlets orNam Sha Shen which has a light brownsurface with a long and fine root Wild CL (wCL) collectedfrom an alpine district of Gangwondo and cultivated CL(cCL) from Yeongcheon of Gyeongbuk in Korea were usedfor this study CL has long been used as traditional forkmedicine to treat asthma phthisis tuberculosis bronchitisdyspepsia and psychoneurosis in Korea Japan and China[6ndash8] The root of CL which is composed of various active

2 Evidence-Based Complementary and Alternative Medicine

components including tannins saponins polyphenolicsalkaloidss essential oils and steroids [6ndash9] has pharmaco-logical properties including antiobesty antioxidant antimi-crobial anti-inflammatory and immunomodulatory activites[10ndash13] Recent studies have documented the biological activ-ities of CL in which water extracts effectively suppressed thehigh-fat-diet-induced accumulation of neutral fat and totalcholesterol in serum and the liver in the experimental model[13] The ethanol extracts of CL demonstrated very strongantioxidant effects comparable to those of ginseng extract[14] In addition Lee et al (1995) and Lee (2002) reportedthat the aqueous extracts of CL acted on the immune systemin direct and indirect manners and increased the cellularimmune reaction [15 16] Another report showed that 70MeOH extracts of CL increased the number of helper T cells[17] Our study was performed to evaluate the antiobesityactivity of CL from different sources the wild type grownnaturally on the mountain and the cultivated typeThis studyattempted to analyze and compare the potential of wCLand cCL as promising resources for the development of afunctional food to control obesity

2 Materials and Methods

21 Plant Material and Reagents wCL (voucher specimennumber SNBA200505262013 theNational BiospeciesKnowl-edge Information System Korea) produced from the Young-mun Mt Corporation of the Agricultural Association andcCL (voucher specimen number KBNA200408101015 theNational Biospecies Knowledge Information System Korea)obtained from Yeongcheon of Gyeongbuk were used for thisstudy These two products were ground separately and 1 literof distilled water was added to 100 g of each of the groundwCL and cCL samples followed by three extraction at 100∘Cover 4 hours using a shaking extraction methodThe extractswere filtered vacuum-dried and then freeze-dried The finalyields of the extract from the dried samples of wCL and cCLwere 72 respectively

Test kits for measuring the total cholesterol triglycerideandHDL-cholesterol in the serumwere purchased fromAsanPharmaceutical Korea Glucose and all other reagents werepurchased from Sigma USA

22 Preparation of Samples for LC-MS Analysis The powderof water extracts from roots of the wCL (200mg) wasdissolved in 100mL of distilled water which was thenpartitioned three times with the same volume of ethyl acetate[18] The water fraction was evaporated to dryness undera vacuum The dried samples were dissolved in distilledwater (1mgmL) and then subjected to a reverse phase HPLCcolumn

23 HPLC-ESI-MS Analysis The HPLC column used wasan Atlantis T3 RP18 (46 times 150mm 3 120583m waters) equippedwith an Atlantis T3 RP18 guard column (46times20mm 35120583mwaters) The mobile phases were constituted with solventA (distilled water) and solvent B (acetonitrile) The linear

Table 1 Composition of the normal diet and the high-caloriehigh-fat diet (g100 g)

Groups ingredient Normal diet High-caloriehigh-fat dietlowast

Casein 200 290Corn starch 600 100Sucrose mdash 100Corn oil 90 50Cellulose 50 50Lard mdash 350AIN-76 mineral mix 35 35AIN-76 vitamin mix 10 10DL-Methionine 03 03Choline bitartrate 02 02kcal100 g diet 3902 4580Calorie from fat () 115 350lowastHigh-caloriehigh-fat diet was modified from the AIN-76 dietary compo-sition [21]

gradient program was as follows 0ndash60min 5ndash50 B 60ndash90min 50ndash70 B 90ndash120min 70ndash100 B running at aconstant flow rate of 02mLmin (P680 Pump DIONEX)The injection volume of the sample was 10 120583L for everyinjection A UVD340U diode array detector performing thewavelength scanning from 210 to 365 nm and an ASI-100automatic injector were usedThe operating conditions of theESI ion source (Ion Sense Tokyo Japan) coupled with a JMS-T100TD (AccuTOF-TLC) in the positive ion modes were adesolating chamber at 250∘C and an orifice at 180∘CThe firstorifice lens was set to 85V and the ring lens voltage was 10VThe TOF-MS was set with a peak voltage of 1500V with adetector voltage of 2200V The nebulizing and desolvatinggases were set to 10 Lmin and 15 Lmin respectively

24 Animals and Treatment Six-week-old male Sprague-Dawley rats which is a good animal model for studying theobesity in humans [19 20] were obtained from Samtako(Osan Korea) and raised under constant conditions (tem-perature 20 plusmn 2∘C humidity 40ndash60 lights 12 hours oflightdark cycle) for 7 days Body weights ranged between200 and 210 gThe rats were provided commercially availablenormal diet (Jongang Lab Animal Seoul Korea) prior to thedietary manipulation All of the samples were administeredat a specific time

During the study period two diets were used the normaldiet or the high-caloriehigh-fat-diet which modified theAIN-76 dietary composition [21] as shown inTable 1 A high-caloriehigh-fat-diet which is commonly used in nutritionalexperiments as a strategy to induce fat deposition andoverweight conditions in SD rat [22 23] was applied forsix weeks to induce obesity The body weight of these ratsincreased more than 125 compared to that of the rats withnormal dietThe rats of the normal test group were fed with anormal diet (ND) for six weeks while the rats of the controlgroup were fed with a high-caloriehigh-fat-diet (HFD) forsix weeks to elicit diet-induced obesity

Then the animals were subdivided into 11 groups (119899 =10)mdashND ND + wCL (900mgkg) ND + cCL (900mgkg)HFD HFD + wCL (100 300 or 900mgkg) HFD + cCL

Evidence-Based Complementary and Alternative Medicine 3

Table 2 Classification of experimental groups

Group TreatmentND Normal dietND + wCL 900mgkg ND + wild Codonopsis lanceolata 900mgkgdayND + cCL 900mgkg ND + cultivated Codonopsis lanceolata 900mgkgdayHFD High-caloriehigh-fat dietHFD + sibutramine HFD + sibutramine 75 11mgkgdayHFD + wCL 100mgkg HFD + wild Codonopsis lanceolata 100mgkgdayHFD + wCL 300mgkg HFD + wild Codonopsis lanceolata 300mgkgdayHFD + wCL 900mgkg HFD + wild Codonopsis lanceolata 900mgkgdayHFD + cCL 100mgkg HFD + cultivated Codonopsis lanceolata 100mgkgdayHFD + cCL 300mgkg HFD + cultivated Codonopsis lanceolata 300mgkgdayHFD + cCL 900mgkg HFD + cultivated Codonopsis lanceolata 900mgkgday

(100 300 or 900mgkg) group and HFD + sibutraminemdashand orally administered with CL or sibutramine once perday for another 6 weeks (Table 2) [23] All experimentalprocedures were carried out in accordance with the protocolthat was approved by the Institutional Animal Care and UseCommittee guideline of Kyung Hee University

25 Specimen Collection Blood was collected from theabdominal aorta of the rat anesthetized using CO

2gas after

fasting for 12 hours on the last day of the 6-week feedingperiod The collected blood was processed using a micro-centrifuge method and the serum was stored in a freezer atminus70∘C The retroperitoneal epididymal and brown fat padswere removed rinsed with phosphate-buffered saline andthen weighed

26 Blood Sampling and Plasma Assay The total cholesterol(TC) triglyceride (TG) and high density lipoprotein (HDL)-cholesterol of the serum were measured using a reagent kit(Asan Pharm Hwaseong Korea) based on the enzymaticcolorimetric method The level of low density lipoprotein-(LDL-) cholesterol was determined using the Friedewaldformula where LDL-cholesterol = TC minus HDL-cholesterol minus(TG5)

27 Cell Culture and Differentiation 3T3-L1 cells were cul-tured to confluence in 6 plates in Dulbeccorsquos Modified EaglersquosMedium (DMEM) supplemented with 10 (vv) Bovine CalfSerum (BCS) On the second day of postconfluence (desig-nated as day 0) the cells were induced to differentiate DMEMsupplemented with 10 (vv) FBS (Fetal Bovine Serum)1 120583M dexamethasone 05mM isobutylmethylxanthine and5 120583gmL insulin After 48 hours themediawere replacedwithDMEMsupplementedwith 10FBS and 1120583gmL insulinThecells were subsequently refed every 48 hours with DMEMsupplemented with 10FBS

28 Oil Red O Staining 3T3-L1 cells were washed twicewith ice-cold phosphate-buffered saline (PBS) fixedwith 10formalin and then stained with 05 Oil Red O

29 Reverse Transcriptase-PCR Analysis of CEBP120572 andPPAR120574 Total RNA was isolated from cells using the Trizolreagent One microgram of total RNA was used for single

strand cDNA synthesis Reverse transcription was performedat 30∘C for 10min 42∘C for 30min and 99∘C for 5minThe following primers were used CCAATenhancer bindingprotein-alpha (CEBP120572) forward 5-CGCAAGAGCCGAG-ATAAAGC-3 and reverse 5-CACGGCTCAGCTGTTC-CA-3 peroxisome proliferator-activated receptor-gamma(PPAR120574) forward 5-CGCTGATGCACTGCCTATGA-3and reverse 5-AGAGGTCCACAGAGCTGATTCC-3 TheCEBP120572 and PPAR120574 amplifications were performed bydenaturing at 94∘C for 1min annealing at 54∘C for 1minextending at 72∘C for 30 sec for 30 cycles administering afinal extension of ethidium bromide and then visualizingunder a UV light

210 Statistical Analysis The results of the tests are presentedas the mean plusmn SD values for each test group using theStatistical Package for the Social Sciences (SPSS) The statis-tical significance between test groups was analyzed using aone-way ANOVA method Differences were analyzed usingDuncanrsquos Multiple Range Test at the 119875 lt 005 level theirsignificances were also analyzed

3 Results

31 Analysis of the Wild Codonopsis lanceolata ExtractFigure 1 shows the HPLC chromatogram and the total ioncurrent chromatogram of the total extract of wCL fromthe HPLC-UVESI-MS analysis The protonated molecu-lar ion of wCL (119898119911 1191 ([M + H]+)) was detected at11min as a codonoposide a compound known to be oneof the active components of CL [24] In addition spinas-terol cycloartenol and taraxerone were identified at 77min98min and 106min respectively [25] These compoundshave already been reported as chemical constituents of thisplant The extracts used in this study were standardized to acodonoposide (yield 03)

32 Body Weight and Weight Gain Table 3 presents theinfluences of wCL and cCL on weight the ingested dietamount and the food efficiency of the rats fed with the dietThere were no particular differences between the test groupsfed with the normal diet (ND) and with the normal dietsupplemented with CL (ND + wCL or cCL 900mgkg)


Table 3 Effects of water extracts of wild and cultivated Codonopsis lanceolata on body weight food efficiency ratio and food intake in SDrats fed with normal or high-fat diets

Group Body weight (g) Food efficiency ratio1 Food intake (gday)Initial Final

ND 2137 plusmn 1011 48160 plusmn 3568

021 plusmn 003

5724 plusmn 817

ND + wCL 900mgkg 2136 plusmn 897 47340 plusmn 2638

017 plusmn 005lowast

5514 plusmn 1006

ND + cCL 900mgkg 2136 plusmn 811 48420 plusmn 2615

022 plusmn 004 5747 plusmn 1233

HFD 2136 plusmn 789 54680 plusmn 3684lowastlowast



HFD + Sibutramine 2136 plusmn 787 47400 plusmn 3020

014 plusmn 003lowastlowast


HFD + wCL 100mgkg 2136 plusmn 766 51622 plusmn 3273lowast

021 plusmn 005

5260 plusmn 542


022 plusmn 007 4992 plusmn 1157

HFD + wCL 900mgkg 2136 plusmn 770 49975 plusmn 3032



HFD + cCL 100mgkg 2136 plusmn 755 50420 plusmn 3120


4885 plusmn 438


022 plusmn 004 4737 plusmn 650

HFD + cCL 900mgkg 2136 plusmn 705 52075 plusmn 2252lowast


5028 plusmn 909

1Food efficiency ratio (FER) = increased body weight (g)Food intake (g)Values are the means plusmn SD (119899 = 10) lowast119875 lt 005 lowastlowast119875 lt 001 versus ND group 119875 lt 005 119875 lt 001 versus HFD group

(min)

(min)

CodonoposideMW 1190

120572-spinasterolMW 412

CycloartenolMW 329

TaraxeroneMW 424

OO O

O

O

O

O

O

O

OHHO

HO

OH

OH

OHOH

O

OH

OHO

OH

HOHO

O

H

H

H

HOHO H

H

H

OH

H

H

H

H

(mAU

)In

tens

ity

TIC

UV 210nm

20 40 60 80 100 1200

11min

77min

98min

106min

Figure 1 HPLC chromatogram and total ion current chromatogram of the crude extract of wCL by LC-UV-ESI-MS

However the rats in the test group fed with the high-caloriehigh-fat-diet (HFD) showed a significant increasein their weight compared to the test group fed with thenormal diet (ND) (119875 lt 001) In addition the amountof daily ingested calories in the control group fed with thehigh-caloriehigh-fat-diet (HFD) represented a significantlyhigher level compared to that of the normal diet (ND)

Interestingly the amount of ingested diet in the groupsfed with the high-fat-diet supplemented with wCL (HFD +wCL 900mgkg) showed a slight decrease over the high-caloriehigh-fat-diet (HFD) (119875 lt 001) There was alsoa significant decrease (119875 lt 001) in weight at the highconcentration (900mgkg) of wCL extracts while the low(100mgkg) andmedium (300mgkg) concentrations of wCL


Table 4 Effects of water extracts of wild and cultivated Codonopsis lanceolata on retroperitoneal epididymal brown fat and total abdominalfat in SD rats fed with normal or high-fat diets

Group Retroperitoneal (g) Epididymal (g) Brown fat (g) Total abdominal (g)ND 621 plusmn 330

940 plusmn 400


ND + wCL 900mgkg 589 plusmn 169

976 plusmn 263


ND + cCL 900mgkg 624 plusmn 120

995 plusmn 336


HFD 1101 plusmn 179lowastlowast


044 plusmn 010 2578 plusmn 367lowastlowast

HFD + Sibutramine 681 plusmn 154

1116 plusmn 389

033 plusmn 007 1860 plusmn 286lowast

HFD + wCL 100mgkg 888 plusmn 120lowast










HFD + cCL 100mgkg 961 plusmn 228lowast











Values are means plusmn SD (119899 = 10) lowast119875 lt 005 lowastlowast119875 lt 001 versus ND group 119875 lt 005 119875 lt 001 versus HFD group

extracts and the high concentration of cCL extracts showedno significant changes in weight compared to the high-caloriehigh-fat-diet (HFD) group For the HFD + cCLgroups there were some changes in the final weight at the lowand medium concentration groups but there were increasesin the food efficiency and no decreases in the ingestedamount Therefore it can be assumed that the results arerelated to dehydration due to the increase in fat and thedecrease in muscle rather than the decrease in body weight

33 Influences on the Food Efficiency To investigate the effectof test samples (sibutramine as positive control and CL) onobesity changes in body weights and ingested food amountswere observed and the food efficiencies were calculated fromthe equation outlined below (Table 3) The body weight wasmeasured every day and the ingested food was calculatedby measuring the amount of remaining food where foodefficiency ratio (FER) = increased body weight (g)foodintake (g)

According to this food efficiency ratio equation a changein body weight is themost important factor affecting the foodefficiency ratio as there is no large change in the amount ofingested food Thus it is possible to apply the food efficiencyratio as a scale of obesity and to consider that a small valuefor the food efficiency ratio is also an effective parameter topredict the avoidance of obesity When verification of diet-induced obesity using this food efficiency ratio was applied inthis study it was possible to observe that upon administrationof CL (ie wCL or cCL) for six weeks obesity was reducedin a concentration-dependent manner to the levels recordedin the normal diet (ND) group The high-capacity wCL900mgkg group showed the results from the ND group andthe HFD group significantly lower FER This as showingthe proper diet inhibition from all of the normal diet or thehigh calories diet is regarded to be a necessary result indiet control for keeping the standard weight In addition thegroups treated with the highest concentrations (900mgkg)of wCL and cCL demonstrated a decrease in the level ofobesity It can be suggested that the high concentration of CLis effective in reducing obesity

34 Plasma Lipid Levels and Fat Pad The obese rats with noCL treatment showed an increase in the weight of the total

fat in the abdominal cavity and in the weight of the retroperitoneum and around the epididymis in the abdominalcavity However the obese rats treated with the CL (ie wCLor cCL) showed a significant latency in the increase in weightof these fat tissues In particular the group treated with wCL(HFD + wCL) had a more efficient control of body weightincrease in a concentration-dependent manner comparedto that of the group treated with cCL (HFD + cCL) Thegroup treated with high concentrations (900mgkg) of wCLshowed themost significant effects on theweight of fat tissueswith the retroperitoneal fat tissues decreasing approximately75 and the weight of epididymal fat tissues decreasingapproximately 83 compared to the high-caloriehigh-fat-diet (HFD) control group The group treated with highconcentrations (900mgkg) of wCL also demonstrated adecrease of approximately 77 in abdominal fat levels andan increase of approximately 145 in brown adipose tissuelevels which facilitates thermal production (Table 4) Amongthe test groups treatedwithCL (HFD+CL) the group treatedwith the medium concentration (600mgkg) showed thesmallest effects To investigate the effects of wCL on the ratsfed with the high-caloriehigh-fat-diet (HFD) in more detailthe effects on serum triglycerides (TG) total cholesterol (TC)and high density lipoprotein-(HDL-) cholesterol levels in therats were measured (Figure 2) Additionally the serum fatconcentrations of the rats with diet-induced obesity weremeasured From the results of the measurements the HFDcontrol group (ie not treated with CL) showed high valuesin serum TG and TC compared to those of the normal diet(ND) group It was evident that the HFD resulted in obesityand high cholesterol levels in rats subjected to this diet Theadministration of CL significantly suppressed the increase ofthese levels In the case of the TG levels these effects wereobserved to be concentration dependent

35 T3-L1 Cells Differentiation Adipocytes adjust their owngrowth and development not only for lipid metabolism butalso for the performance of functions tomaintain a consistentsupply of energy in the body [26]

In this study we observed the effects of wCL on adipocytedifferentiation by a cell-based in vitro experiment with Oil


900

900 900

900

100

100

100

80

60

40

20

0

300

300 (mgkg)

ND HFD

Seru

m to

tal c

hole

stero

l (m

gdL

)

lowast

75

11minus minus

(a)

900

900 900

900

100

100

60

40

20

0

300

300 (mgkg)

ND HFD

lowast

75

11

Seru

m tr

igly

cerid

es (m

gdL

)

minus

minus minus

(b)

900 900 900

900

100

100

60

40

20

0

300

300 (mgkg)

ND HFD

75

11

Seru

m H

DL-

chol

este

rol (

mg

dL)

ControlSibutramine

wCLcCL

minus minus

(c)

900

900900

900

100

100

30

20

10

0

300

300

(mgkg)

ND HFD75

11

ControlSibutramine

wCLcCL

Seru

m L

DL-

chol

este

rol (

mg

dL)

lowast

lowast

minus minus

(d)

Figure 2 Effects of the wild and cultivated Codonopsis lanceolata extracts on (a) serum total cholesterol (b) serum triglycerides (c) serumhigh density lipoprotein (HDL)-cholesterol (d) serum low density lipoprotein-(LDL-) cholesterol levels (mgdL) in rats Values are means plusmnSD (119899 = 10) lowast119875 lt 005 lowastlowast119875 lt 001 versus ND group 119875 lt 005 119875 lt 001 versus HFD group

RedO dyeThe results shown in Figure 3 confirmed that 3T3-L1 fat generation is suppressed with treatment of wCL in aconcentration-dependent manner A similar concentration-dependent effect ofwCLon the triglyceride levels in the bloodsamples confirmed the strong relationship between adipocytedifferentiation and triglyceride levels in the blood (Figure 2)Because the level of triglycerides in serum increases withincreases in the obesity index and plays an important role inlipid metabolism it may be considered to be a major deter-minating parameter for obesity To investigate the possiblemolecular mechanism of wCL on the suppression of obesitythe levels of CCAATenhancer binding protein 120572 (CEBP120572)mRNA and peroxisome proliferator-activated receptor 120574(PPAR120574) mRNA were monitored in 3T3-L1 cells treated withincreasing concentrations of wCL It was observed that theexpression of CEBP120572 mRNA one of the most importanttranscription factors in cell differentiation decreased withincreasing concentrations of wCL and the expression ofPPAR120574 mRNA which plays an important role in promotingstorage of fatty acids in adipose tissue also decreased withwCL treatments in a concentration-dependent manner

4 Discussion

It is widely accepted that the levels of saturated fat andcholesterol in the diet increase the level of serum cholesterolwhile a diet with a low fat content decreases the level ofserum cholesterol There have been tremendous efforts tofind efficient food sources from traditional herbal medicinesthat can be used to optimize the level of serum cholesterolthereby controlling obesity Ginseng is one of the mostpromising candidates for a functional food of this type Ithas been reported that the saponin in ginseng increased theabsorption of LDL in the liver and facilitated the removal rateof abnormally increased VLDL by the intake of high levelsof cholesterol from the diet [27] It has also been reportedthat the decrease in the level of blood cholesterol by theadministration of saponin derived from ginseng was mainlydependent on the decrease in the concentrations of VLDLand LDL in serum In this present study it was also assumedthat the significant decreases in the total blood cholesterolof all groups fed with HFD and treated with wCL were dueto the action of the saponins in the wCL as saponins have


MDI minus

minus

+ + + +

wCL (mgmL) 0 03 05 07

(a)

minus

minusMDI

wCL (mgmL)+

0

0

20

40

60

80

100

120

+

03

+

05

+

07

Relat

ive l

ipid

cont

ent (

)

lowast

lowastlowast

(b)

CEBP120572

120573-actin

PPAR120574

minus

minusMDI

wCL (mgmL)+

0

+

03

+

05

+

07

(c)

Figure 3 The effect of wCL on 3T3-L1 adipocyte differentiation (a) 3T3-L1 cells were differentiated with MDI in the absence or presence ofwCL (0 03 05 07mgmL) for eight days followed by measurement of lipid contents by Oil Red O staining (b) Stained oil droplets weredissolved with isopropanol and quantified by spectrophotometric analysis at 510 nm The results were represented as relative lipid contentslowast

119875 lt 005 lowastlowast119875 lt 001 by SPSS compared to MDI-treated cells (c) Expression of CEBP120572 and PPAR120574mRNA from 3T3-L1 cells as describedin (a) was measured by RT-PCR

been reported as active components of CL [28] Althoughthe uniform effects on the levels of serum cholesterol andlipids were obtained in the group treated with the highestconcentration level of wCL (HFD + wCL 900mgkg) therewere small improvements in the fat and blood lipid levels inthe group treated with cCL (HFD + cCL) The reason thatwCL water extracts showed more effective results than cCLextracts is likely due to differences in the content and types ofactive ingredients between wCL and cCL products

Mitochondrial120573-oxidation of long-chain fatty acids is themajor source of energy Prior to undergoing 120573-oxidation inmitochondrial matrix the long-chain fatty acyl-CoAmust betransferred from cytosol into matrix The lipid metabolismrelated many enzymes in this process The present studyhas investigated the effects of wCL on lipid and cholesterolmetabolisms in normolipidemic and hyperlipidemic rat Wewere able to get the results wCL inhibiting activity of fatoxidation

The cholesterol lowering of property wCL could bedue to increased excretion of cholesterol and bile acidsThese observed effects can be attributed to the presence ofphytosterol in wCL as phytosterols possesse greater affinityfor micelles than cholesterol and reduce incorporation ofcholesterol in micelles [29] We can see from the configu-ration that the phytochemical components of this wCL arecomposed of the enzyme controlling largely physiologicalvitality of adipose metabolism and joining matters

The effect of CL extract on obesity control does not seemto be related to a direct inhibition of the neurotransmitter cas-cade the main mechanism of sibutramine and a serotonin-norepinephrine reuptake inhibitor The treatment with wCLresulted in the control of serum lipid levels a decrease inthe content of the total body fat and a significant increasein the level of brown adipose tissue However sibutramine-treated models (HFD + sibutramine) have shown that theobesity control effect does not accompany an increase in the


brown adipose tissue that is known to have high capabilityin fat burning and fatty acid oxidation Direct action of CLon the lipid levels in serum rather than a central controlof appetite could exclude various side effects in the centralnervous system (CNS) found in antiobesity agents such assibutramine

Growth of adipose tissues is broadly divided into twostages the increase in fat cell size and the differentiation of theadipocyte from the preadipocyte [30] During differentiationfrom precursor adipocytes into fully mature adipocyteseither ecologically or biochemically differentiating factors areactivated which are important for regulating the adipocytegenes [30 31]

Various transcription factors like PPAR120574 and CEBPfamily are involved during adipocyte differentiation Amongthem the expression of PPAR120574 and CEBP120572 in 3T3-L1 whichis a wellknown and frequently used preadipocyte cell linefor in vitro adipocyte differentiation [32] is upregulated[33] PPAR120574 and CEBP120572 the key adipogenic and lipogenictranscriptional regulators are crucial to the regulation ofobesity and adipocyte differentiation [34] Recently studieshave shown that PPAR120574 is necessary and sufficient to promoteadipogenesis and that CEBP120572 is influential in maintainingthe expression of PPAR120574 [35ndash37] The results of our studydemonstrate that the treatment with wCL extract suppressesthe accumulation of triglycerides within the cells and thedifferentiation of 3T3-L1 preadipocytes into adipocytes TotalCL group significantly reduced fat accumulation by inhibitingadipogenic signal transcriptional factors such as PPAR120574and CEBP120572 mRNA which functions via AMPK (51015840 AMP-activated protein kinase or 51015840adenosine monophosphate-activated protein kinase) signaling in vitro In other wordsthe concentration-dependent suppression of CEBP120572 andPPAR120574mRNA expression in CL-treated cells represented oneof the possible mechanisms of the suppression of 3T3-L1 cellsdifferentiation

AMPK is an enzyme that plays a role in cellular energyhomeostasisThenet effect of AMPK activation is stimulationof hepatic fatty acid oxidation and ketogenesis and inhi-bition of cholesterol synthesis lipogenesis and triglyceridesynthesis [38] AMPK and PPAR120574 appear to be involvedin adipocyte differentiation and maturation and thus canbe potential drug targets for the treatment of obesity wCLsignificantly inhibited the expression levels of CEBP120572 andPPAR120574 two master regulators of adipogenesis (Figure 3)indicating that wCL might inhibit 3T3-L1 differentiation viasuppressing the expression of adipogenesis-related transcrip-tion factors and markers Meanwhile PPAR120574 transcriptionalactivity was reduced supporting the wCL downregulatedPPAR120574 expression as well as its transcriptional activityAMPK phosphorylates the transcriptional coactivator p300and induces its interaction with PPAR120574 [33] It was supposedthat wCL might suppress PPAR120574 transcriptional activity viaactivating AMPK and phosphorylating the transcriptionalcoactivators and hence leading to the inhibition of theirabilities to interact with PPAR120574

In conclusion we can observe that wCL improved therestraint of excessive adipose formation and accumulation bythese matters The potential of CL as a promising source for

the development of a functional food which can be efficientlyused to control obesity should be appreciated and evaluatedwith more detailed molecular studies

Acknowledgment

This work was supported in part by Grant 106069-3 from theAgricultural RampD Promotion Center

References

[1] A L Sajem and K Gosai ldquoTraditional use of medicinal plantsby the Jaintia tribes in North Cachar Hills district of Assamnortheast Indiardquo Journal of Ethnobiology and Ethnomedicinevol 2 article 33 2006

[2] R N Jadeja M C Thounaojam Ansarullah R V Devkarand A V Ramachandran ldquoA preliminary study on hypolipi-demic effect of aqueous leaf extract of Clerodendron glandulo-sumColebrdquo International Journal of Green Pharmacy vol 3 no4 pp 285ndash289 2009

[3] R N Jadeja M C Thounaojam Ansarullah R V Devkarand A V Ramachandran ldquoClerodendron glandulosum ColebVerbenaceae ameliorates high fat diet-induced alteration inlipid and cholesterol metabolism in ratsrdquo Brazilian Journal ofPharmacognosy vol 20 no 1 pp 117ndash123 2010

[4] R N Jadeja M C Thounaojam Ansarullah V B PatelR V Devkar and A V Ramachandran ldquoProtective effectof Clerodendron glandulosum extract against experimentallyinduced metabolic syndrome in ratsrdquo Pharmaceutical Biologyvol 48 no 12 pp 1312ndash1319 2010

[5] R N Jadeja M C Thounaojam D S Dandekar R V Devkarand A V Ramachandran ldquoClerodendron glandulosumColebextract ameliorates high fat dietfatty acid induced lipotoxicityin experimental models of non-alcoholic steatohepatitisrdquo Foodand Chemical Toxicology vol 48 no 12 pp 3424ndash3431 2010

[6] Z T Wang G Y Ma P F Tu et al ldquoChemotaxonomic studyof Codonopsis (family Campanulaceae) and its related generardquoBiochemical Systematics and Ecology vol 24 no 7-8 pp 809ndash812 1995

[7] M Ichikawa S Ohta N Komoto et al ldquoSimultaneous determi-nation of seven saponins in the roots ofCodonopsis lanceolatabyliquid chromatography-mass spectrometryrdquo Journal of NaturalMedicines vol 63 no 1 pp 52ndash57 2009

[8] K T Lee J ChoiW T Jung J HNamH J Jung andH J ParkldquoStructure of a new echinocystic acid bisdesmoside isolatedfrom Codonopsis lanceolata roots and the cytotoxic activity ofprosapogeninsrdquo Journal of Agricultural and Food Chemistry vol50 no 15 pp 4190ndash4193 2002

[9] C Y Li H X Xu Q B Han and T S Wu ldquoQuality assess-ment of Radix Codonopsis by quantitative nuclear magneticresonancerdquo Journal of Chromatography A vol 1216 no 11 pp2124ndash2129 2009

[10] S Yongxu and L Jicheng ldquoStructural characterization of awater-soluble polysaccharide from the roots of Codonopsispilosula and its immunity activityrdquo International Journal ofBiological Macromolecules vol 43 no 3 pp 279ndash282 2008

[11] S Byeon W Choi E Houng J Lee et al ldquoInhibitory effectof saponin fraction from Codonopsis lanceolata on immunecell-mediatd inflammatlry responesrdquo Archives of PharmacalResearch vol 32 pp 813ndash822 2009


[12] Y S Maeng and H K Park ldquoAntioxidant activity of ethanolextract from dodok (Codonopsis lanceolata)rdquo Korean Journal ofFood Science and Technology vol 23 pp 311ndash316 1991

[13] E G Han I S Sung H G Moon and S Y Cho ldquoEffects ofCodonopsis lanceolata water extract on the level of lipid in ratsfed high fat dietrdquo Journal of the Korean Society of Food Scienceand Nutrition vol 27 pp 940ndash944 1998

[14] RQisheng Y Xiongying S Xinrong andCHuangshi ldquoChem-ical constituents fromCodonopsis lanceolatardquoZhongcaoyao vol36 pp 1773ndash1775 2005

[15] Y J Lee JM Kim andYM Jung ldquoEffect ofCodonopsis pilosulson the cellular immunityrdquo The Korean Journal of VeterinaryResearch vol 19 pp 273ndash279 1995

[16] J H Lee ldquoImmunostimulative effect of hot-water extract fromCodonopsis lanceolata on lymphocyte and clonal macrophagerdquoKorean Journal of Food Science and Technology vol 34 pp 732ndash736 2002

[17] Y G Lee J Y Kim J Y Lee et al ldquoRegulatory effectsof Codonopsis lanceolata on macrophage-mediated immuneresponsesrdquo Journal of Ethnopharmacology vol 112 no 1 pp180ndash188 2007

[18] J Atzrodt VDerdauWHolla andM Sandvoss ldquoThe synthesisof selected phase II metabolitesmdashO-glucuronides and sulfatesof drug development candidatesrdquo Arkivoc vol 3 pp 257ndash2782012

[19] S P Xu X YMao X Cheng and B Chen ldquoAmeliorating effectsof casein glycomacropeptide on obesity induced by high-fat dietin male Sprague-Dawley ratsrdquo Food and Chemical Toxicologyvol 56 pp 1ndash7 2013

[20] H Inoue K Kobayashi-Hattori Y Horiuchi Y Oishi SArai and T Takita ldquoRegulation of the body fat percentage indevelopmental-stage rats by methylxanthine derivatives in ahigh-fat dietrdquo Bioscience Biotechnology and Biochemistry vol70 no 5 pp 1134ndash1139 2006

[21] P G Reeves F H Nielsen and G C Fahey GC ldquoAmericanInstitute of Nutrition Report of the American Institute ofNutrition ad hoc Committee on Standards for NutritionalStudiesrdquoThe Journal of Nutrition vol 107 pp 1340ndash1348 1977

[22] C J Zhou S H Huang J Q Liu et al ldquoSweet tea leaves extractimproves leptin resistance in diet-induced obese ratsrdquo Journalof Ethnopharmacology vol 145 pp 386ndash392 2013

[23] W S Jang and S Y Choung ldquoAntiobestiy effects of theethanol extract of Laminaria japonica areshoung in high-fat-diet-induced obese ratrdquo Evidence-Based Complementary andAlternativeMedicine vol 2013 Article ID 492807 17 pages 2013

[24] J S Lee M K Lee T Y Ha et al ldquoSupplementation ofwhole persimmon leaf improves lipid profiles and suppressesbody weight gain in rats fed high-fat dietrdquo Food and ChemicalToxicology vol 44 no 11 pp 1875ndash1883 2006

[25] M D Gades and J S Stern ldquoChitosan supplementation doesnot affect fat absorption in healthy males fed a high-fat diet apilot studyrdquo International Journal of Obesity vol 26 no 1 pp119ndash122 2002

[26] H L Zhao J S Sim S H Shim Y W Ha S S Kang andY S Kim ldquoAntiobese and hypolipidemic effects of platycodinsaponins in diet-induced obese rats evidences for lipase inhi-bition and calorie intake restrictionrdquo International Journal ofObesity vol 29 no 8 pp 983ndash990 2005

[27] M S So J S Lee and S Y Yi ldquoInduction of Nitric Oxide andcytokines in macrophages by Codonopsis lanceolatardquo KoreanJournal of Food Science and Technology vol 36 pp 986ndash9902004

[28] BH Kang andCN Joo ldquoThe effect of ginseng saponin fractionon low density lipoprotein (LDL) uptake by rat and rabbitliversrdquo Korean Biochemical Journal vol 19 pp 168ndash172 1986

[29] I Ikeda and M Sugano ldquoInhibition of cholesterol absorptionby plant sterols for mass interventionrdquo Current Opinion inLipidology vol 9 no 6 pp 527ndash531 1998

[30] J Y Park and B J Kim ldquoMolecular insights into fat celldifferentiation and functional roles of adipocytokinesrdquo Journalof Korean Society of Endocrinology vol 17 pp 1ndash9 2002

[31] F M Gregoire C M Smas and H S Sul ldquoUnderstandingadipocyte differentiationrdquo Physiological Reviews vol 78 no 3pp 783ndash809 1998

[32] H Green and O Kehinde ldquoAn established preadipose cell lineand its differentiation in culture II Factors affecting the adiposeconversionrdquo Cell vol 5 no 1 pp 19ndash27 1975

[33] F Karadeniz M Z Karagozlu S Y Pyun and S K KimldquoSulfation of chitosan oligomers enhances their anti-adipogeniceffect in 3T3-L1 adipocytesrdquoCarbohydrate Polymers vol 86 no2 pp 666ndash671 2011

[34] A Ejaz D Wu P Kwan and M Meydani ldquoCurcumin inhibitsadipogenesis in 3T3-L1 adipocytes and angiogenesis and obesityinC57BLmicerdquo Journal ofNutrition vol 139 no 5 pp 919ndash9252009

[35] Y Barak M C Nelson E S Ong et al ldquoPPAR120574 is required forplacental cardiac and adipose tissue developmentrdquo MolecularCell vol 4 no 4 pp 585ndash595 1999

[36] H Koutnikova T A Cock M Watanabe et al ldquoCompensationby the muscle limits the metabolic consequences of lipodystro-phy in PPAR120574 hypomorphic micerdquo Proceedings of the NationalAcademy of Sciences of the United States of America vol 100 no2 pp 14457ndash14462 2003

[37] Z D Wu E D Rosen R Brun et al ldquoCross-regulation ofcontrols the transcriptional pathway of CEBP120572 and PPAR120574adipogenesis and insulin sensitivityrdquo Molecules and Cells vol3 pp 151ndash158 1999

[38] WWWinder andDGHardie ldquoAMP-activated protein kinasea metabolic master swith possible roles in type 2 diabetesrdquoAmerican Journal of Physiology vol 277 pp E1ndashE10 1999

Submit your manuscripts athttpwwwhindawicom

Stem CellsInternational

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014


MEDIATORSINFLAMMATION

of


Behavioural Neurology

EndocrinologyInternational Journal of



Disease Markers


BioMed Research International

OncologyJournal of



Oxidative Medicine and Cellular Longevity


PPAR Research

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Immunology ResearchHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

ObesityJournal of



Computational and Mathematical Methods in Medicine

OphthalmologyJournal of


Diabetes ResearchJournal of



Research and TreatmentAIDS


Gastroenterology Research and Practice


Parkinsonrsquos Disease

Evidence-Based Complementary and Alternative Medicine

Volume 2014Hindawi Publishing Corporationhttpwwwhindawicom


components including tannins saponins polyphenolicsalkaloidss essential oils and steroids [6ndash9] has pharmaco-logical properties including antiobesty antioxidant antimi-crobial anti-inflammatory and immunomodulatory activites[10ndash13] Recent studies have documented the biological activ-ities of CL in which water extracts effectively suppressed thehigh-fat-diet-induced accumulation of neutral fat and totalcholesterol in serum and the liver in the experimental model[13] The ethanol extracts of CL demonstrated very strongantioxidant effects comparable to those of ginseng extract[14] In addition Lee et al (1995) and Lee (2002) reportedthat the aqueous extracts of CL acted on the immune systemin direct and indirect manners and increased the cellularimmune reaction [15 16] Another report showed that 70MeOH extracts of CL increased the number of helper T cells[17] Our study was performed to evaluate the antiobesityactivity of CL from different sources the wild type grownnaturally on the mountain and the cultivated typeThis studyattempted to analyze and compare the potential of wCLand cCL as promising resources for the development of afunctional food to control obesity

2 Materials and Methods

21 Plant Material and Reagents wCL (voucher specimennumber SNBA200505262013 theNational BiospeciesKnowl-edge Information System Korea) produced from the Young-mun Mt Corporation of the Agricultural Association andcCL (voucher specimen number KBNA200408101015 theNational Biospecies Knowledge Information System Korea)obtained from Yeongcheon of Gyeongbuk were used for thisstudy These two products were ground separately and 1 literof distilled water was added to 100 g of each of the groundwCL and cCL samples followed by three extraction at 100∘Cover 4 hours using a shaking extraction methodThe extractswere filtered vacuum-dried and then freeze-dried The finalyields of the extract from the dried samples of wCL and cCLwere 72 respectively

Test kits for measuring the total cholesterol triglycerideandHDL-cholesterol in the serumwere purchased fromAsanPharmaceutical Korea Glucose and all other reagents werepurchased from Sigma USA

22 Preparation of Samples for LC-MS Analysis The powderof water extracts from roots of the wCL (200mg) wasdissolved in 100mL of distilled water which was thenpartitioned three times with the same volume of ethyl acetate[18] The water fraction was evaporated to dryness undera vacuum The dried samples were dissolved in distilledwater (1mgmL) and then subjected to a reverse phase HPLCcolumn

23 HPLC-ESI-MS Analysis The HPLC column used wasan Atlantis T3 RP18 (46 times 150mm 3 120583m waters) equippedwith an Atlantis T3 RP18 guard column (46times20mm 35120583mwaters) The mobile phases were constituted with solventA (distilled water) and solvent B (acetonitrile) The linear

Table 1 Composition of the normal diet and the high-caloriehigh-fat diet (g100 g)

Groups ingredient Normal diet High-caloriehigh-fat dietlowast

Casein 200 290Corn starch 600 100Sucrose mdash 100Corn oil 90 50Cellulose 50 50Lard mdash 350AIN-76 mineral mix 35 35AIN-76 vitamin mix 10 10DL-Methionine 03 03Choline bitartrate 02 02kcal100 g diet 3902 4580Calorie from fat () 115 350lowastHigh-caloriehigh-fat diet was modified from the AIN-76 dietary compo-sition [21]

gradient program was as follows 0ndash60min 5ndash50 B 60ndash90min 50ndash70 B 90ndash120min 70ndash100 B running at aconstant flow rate of 02mLmin (P680 Pump DIONEX)The injection volume of the sample was 10 120583L for everyinjection A UVD340U diode array detector performing thewavelength scanning from 210 to 365 nm and an ASI-100automatic injector were usedThe operating conditions of theESI ion source (Ion Sense Tokyo Japan) coupled with a JMS-T100TD (AccuTOF-TLC) in the positive ion modes were adesolating chamber at 250∘C and an orifice at 180∘CThe firstorifice lens was set to 85V and the ring lens voltage was 10VThe TOF-MS was set with a peak voltage of 1500V with adetector voltage of 2200V The nebulizing and desolvatinggases were set to 10 Lmin and 15 Lmin respectively

24 Animals and Treatment Six-week-old male Sprague-Dawley rats which is a good animal model for studying theobesity in humans [19 20] were obtained from Samtako(Osan Korea) and raised under constant conditions (tem-perature 20 plusmn 2∘C humidity 40ndash60 lights 12 hours oflightdark cycle) for 7 days Body weights ranged between200 and 210 gThe rats were provided commercially availablenormal diet (Jongang Lab Animal Seoul Korea) prior to thedietary manipulation All of the samples were administeredat a specific time

During the study period two diets were used the normaldiet or the high-caloriehigh-fat-diet which modified theAIN-76 dietary composition [21] as shown inTable 1 A high-caloriehigh-fat-diet which is commonly used in nutritionalexperiments as a strategy to induce fat deposition andoverweight conditions in SD rat [22 23] was applied forsix weeks to induce obesity The body weight of these ratsincreased more than 125 compared to that of the rats withnormal dietThe rats of the normal test group were fed with anormal diet (ND) for six weeks while the rats of the controlgroup were fed with a high-caloriehigh-fat-diet (HFD) forsix weeks to elicit diet-induced obesity

Then the animals were subdivided into 11 groups (119899 =10)mdashND ND + wCL (900mgkg) ND + cCL (900mgkg)HFD HFD + wCL (100 300 or 900mgkg) HFD + cCL






2gas after








3 Results






ND 2137 plusmn 1011 48160 plusmn 3568

021 plusmn 003

5724 plusmn 817



5514 plusmn 1006


022 plusmn 004 5747 plusmn 1233








021 plusmn 005

5260 plusmn 542


022 plusmn 007 4992 plusmn 1157






4885 plusmn 438





5028 plusmn 909


(min)

(min)

CodonoposideMW 1190


CycloartenolMW 329

TaraxeroneMW 424

OO O

O

O

O

O

O

O

OHHO

HO

OH

OH

OHOH

O

OH

OHO

OH

HOHO

O

H

H

H

HOHO H

H

H

OH

H

H

H

H

(mAU

)In

tens

ity

TIC

UV 210nm

20 40 60 80 100 1200

11min

77min

98min

106min







940 plusmn 400



976 plusmn 263



995 plusmn 336






1116 plusmn 389
































900

900 900

900

100

100

100

80

60

40

20

0

300

300 (mgkg)

ND HFD

Seru

m to

tal c

hole

stero

l (m

gdL

)

lowast

75

11minus minus

(a)

900

900 900

900

100

100

60

40

20

0

300

300 (mgkg)

ND HFD

lowast

75

11

Seru

m tr

igly

cerid

es (m

gdL

)

minus

minus minus

(b)

900 900 900

900

100

100

60

40

20

0

300

300 (mgkg)

ND HFD

75

11

Seru

m H

DL-

chol

este

rol (

mg

dL)

ControlSibutramine

wCLcCL

minus minus

(c)

900

900900

900

100

100

30

20

10

0

300

300

(mgkg)

ND HFD75

11

ControlSibutramine

wCLcCL

Seru

m L

DL-

chol

este

rol (

mg

dL)

lowast

lowast

minus minus

(d)



4 Discussion



MDI minus

minus

+ + + +

wCL (mgmL) 0 03 05 07

(a)

minus

minusMDI

wCL (mgmL)+

0

0

20

40

60

80

100

120

+

03

+

05

+

07

Relat

ive l

ipid

cont

ent (

)

lowast

lowastlowast

(b)

CEBP120572

120573-actin

PPAR120574

minus

minusMDI

wCL (mgmL)+

0

+

03

+

05

+

07

(c)














Acknowledgment


References













































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of





















2gas after








3 Results






ND 2137 plusmn 1011 48160 plusmn 3568

021 plusmn 003

5724 plusmn 817



5514 plusmn 1006


022 plusmn 004 5747 plusmn 1233








021 plusmn 005

5260 plusmn 542


022 plusmn 007 4992 plusmn 1157






4885 plusmn 438





5028 plusmn 909


(min)

(min)

CodonoposideMW 1190


CycloartenolMW 329

TaraxeroneMW 424

OO O

O

O

O

O

O

O

OHHO

HO

OH

OH

OHOH

O

OH

OHO

OH

HOHO

O

H

H

H

HOHO H

H

H

OH

H

H

H

H

(mAU

)In

tens

ity

TIC

UV 210nm

20 40 60 80 100 1200

11min

77min

98min

106min







940 plusmn 400



976 plusmn 263



995 plusmn 336






1116 plusmn 389
































900

900 900

900

100

100

100

80

60

40

20

0

300

300 (mgkg)

ND HFD

Seru

m to

tal c

hole

stero

l (m

gdL

)

lowast

75

11minus minus

(a)

900

900 900

900

100

100

60

40

20

0

300

300 (mgkg)

ND HFD

lowast

75

11

Seru

m tr

igly

cerid

es (m

gdL

)

minus

minus minus

(b)

900 900 900

900

100

100

60

40

20

0

300

300 (mgkg)

ND HFD

75

11

Seru

m H

DL-

chol

este

rol (

mg

dL)

ControlSibutramine

wCLcCL

minus minus

(c)

900

900900

900

100

100

30

20

10

0

300

300

(mgkg)

ND HFD75

11

ControlSibutramine

wCLcCL

Seru

m L

DL-

chol

este

rol (

mg

dL)

lowast

lowast

minus minus

(d)



4 Discussion



MDI minus

minus

+ + + +

wCL (mgmL) 0 03 05 07

(a)

minus

minusMDI

wCL (mgmL)+

0

0

20

40

60

80

100

120

+

03

+

05

+

07

Relat

ive l

ipid

cont

ent (

)

lowast

lowastlowast

(b)

CEBP120572

120573-actin

PPAR120574

minus

minusMDI

wCL (mgmL)+

0

+

03

+

05

+

07

(c)














Acknowledgment


References













































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of



















ND 2137 plusmn 1011 48160 plusmn 3568

021 plusmn 003

5724 plusmn 817



5514 plusmn 1006


022 plusmn 004 5747 plusmn 1233








021 plusmn 005

5260 plusmn 542


022 plusmn 007 4992 plusmn 1157






4885 plusmn 438





5028 plusmn 909


(min)

(min)

CodonoposideMW 1190


CycloartenolMW 329

TaraxeroneMW 424

OO O

O

O

O

O

O

O

OHHO

HO

OH

OH

OHOH

O

OH

OHO

OH

HOHO

O

H

H

H

HOHO H

H

H

OH

H

H

H

H

(mAU

)In

tens

ity

TIC

UV 210nm

20 40 60 80 100 1200

11min

77min

98min

106min







940 plusmn 400



976 plusmn 263



995 plusmn 336






1116 plusmn 389
































900

900 900

900

100

100

100

80

60

40

20

0

300

300 (mgkg)

ND HFD

Seru

m to

tal c

hole

stero

l (m

gdL

)

lowast

75

11minus minus

(a)

900

900 900

900

100

100

60

40

20

0

300

300 (mgkg)

ND HFD

lowast

75

11

Seru

m tr

igly

cerid

es (m

gdL

)

minus

minus minus

(b)

900 900 900

900

100

100

60

40

20

0

300

300 (mgkg)

ND HFD

75

11

Seru

m H

DL-

chol

este

rol (

mg

dL)

ControlSibutramine

wCLcCL

minus minus

(c)

900

900900

900

100

100

30

20

10

0

300

300

(mgkg)

ND HFD75

11

ControlSibutramine

wCLcCL

Seru

m L

DL-

chol

este

rol (

mg

dL)

lowast

lowast

minus minus

(d)



4 Discussion



MDI minus

minus

+ + + +

wCL (mgmL) 0 03 05 07

(a)

minus

minusMDI

wCL (mgmL)+

0

0

20

40

60

80

100

120

+

03

+

05

+

07

Relat

ive l

ipid

cont

ent (

)

lowast

lowastlowast

(b)

CEBP120572

120573-actin

PPAR120574

minus

minusMDI

wCL (mgmL)+

0

+

03

+

05

+

07

(c)














Acknowledgment


References













































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of



















940 plusmn 400



976 plusmn 263



995 plusmn 336






1116 plusmn 389
































900

900 900

900

100

100

100

80

60

40

20

0

300

300 (mgkg)

ND HFD

Seru

m to

tal c

hole

stero

l (m

gdL

)

lowast

75

11minus minus

(a)

900

900 900

900

100

100

60

40

20

0

300

300 (mgkg)

ND HFD

lowast

75

11

Seru

m tr

igly

cerid

es (m

gdL

)

minus

minus minus

(b)

900 900 900

900

100

100

60

40

20

0

300

300 (mgkg)

ND HFD

75

11

Seru

m H

DL-

chol

este

rol (

mg

dL)

ControlSibutramine

wCLcCL

minus minus

(c)

900

900900

900

100

100

30

20

10

0

300

300

(mgkg)

ND HFD75

11

ControlSibutramine

wCLcCL

Seru

m L

DL-

chol

este

rol (

mg

dL)

lowast

lowast

minus minus

(d)



4 Discussion



MDI minus

minus

+ + + +

wCL (mgmL) 0 03 05 07

(a)

minus

minusMDI

wCL (mgmL)+

0

0

20

40

60

80

100

120

+

03

+

05

+

07

Relat

ive l

ipid

cont

ent (

)

lowast

lowastlowast

(b)

CEBP120572

120573-actin

PPAR120574

minus

minusMDI

wCL (mgmL)+

0

+

03

+

05

+

07

(c)














Acknowledgment


References













































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of

















900

900 900

900

100

100

100

80

60

40

20

0

300

300 (mgkg)

ND HFD

Seru

m to

tal c

hole

stero

l (m

gdL

)

lowast

75

11minus minus

(a)

900

900 900

900

100

100

60

40

20

0

300

300 (mgkg)

ND HFD

lowast

75

11

Seru

m tr

igly

cerid

es (m

gdL

)

minus

minus minus

(b)

900 900 900

900

100

100

60

40

20

0

300

300 (mgkg)

ND HFD

75

11

Seru

m H

DL-

chol

este

rol (

mg

dL)

ControlSibutramine

wCLcCL

minus minus

(c)

900

900900

900

100

100

30

20

10

0

300

300

(mgkg)

ND HFD75

11

ControlSibutramine

wCLcCL

Seru

m L

DL-

chol

este

rol (

mg

dL)

lowast

lowast

minus minus

(d)



4 Discussion



MDI minus

minus

+ + + +

wCL (mgmL) 0 03 05 07

(a)

minus

minusMDI

wCL (mgmL)+

0

0

20

40

60

80

100

120

+

03

+

05

+

07

Relat

ive l

ipid

cont

ent (

)

lowast

lowastlowast

(b)

CEBP120572

120573-actin

PPAR120574

minus

minusMDI

wCL (mgmL)+

0

+

03

+

05

+

07

(c)














Acknowledgment


References













































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of

















MDI minus

minus

+ + + +

wCL (mgmL) 0 03 05 07

(a)

minus

minusMDI

wCL (mgmL)+

0

0

20

40

60

80

100

120

+

03

+

05

+

07

Relat

ive l

ipid

cont

ent (

)

lowast

lowastlowast

(b)

CEBP120572

120573-actin

PPAR120574

minus

minusMDI

wCL (mgmL)+

0

+

03

+

05

+

07

(c)














Acknowledgment


References













































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of























Acknowledgment


References













































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of

















































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of
















antiobesity effect of codonopsis lanceolata in high-calorie/high-fat-diet-induced obese rats

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