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DOI:10.3736/ jcim20110517
http:// ww w.jcimjournal.com
Kumar S , Kum ar V , P rakash O.A ntidiabe tic and
hypo lipidemic activities of Dil lenia indica extr act in
diabetic ra ts.J Chin Integr Med .2011;9(5):570-
574.
Kumar S , Kumar V , P raka sh O.五桠果提取物对糖尿
病大鼠的降血糖及降血脂作用.中西医结合学报.
2011;9(5):570-574.
Received Janua ry 24 , 2011; accepted February 16 ,
2011;published online May 15 , 2011.
Full-tex t LinkOut at PubMed.Journal title in PubMed:
Zhong X i Y i J ie He X ue Bao.
Correspondence:Vipin Kumar , PhD , Associate Professor;
Tel:+91-1744-239617;E-mail:vipbhardhwaj@rediffmail.
com
曹和欣 , 何立群 , 沈雅静.糖尿病高血脂所致大鼠肾组织 SOD
和 GSH 及 MDA 的变化.中西医结合学报.2004;2(1):36-
38.
Cao HX , He LQ , Shen YJ.Changes of SOD , GSH and MDA
in renal tissues o f diabetic hyper lipidemia r ats.J Chin Integr
Med .2004;2(1):36-38.
Full text available at http:// ww w.jcimjournal.com/ FullText 2.
a spx?ar ticleID=244
朱芸 , 刘金荣 , 张伟.降血糖药用植物资源概述.中西医结合
学报.2004;2(1):67-68.
Zhu Y , Liu JR , Zhang W.An over view of medicinal plant
resource fo r decreasing blo od gluco se.J Chin Integr Med .
2004;2(1):67-68.
Full text available at http:// ww w.jcimjournal.com/ FullText 2.
a spx?ar ticleID=255
More related articles at http://www.jcimjournal.com/ FullText 2.
a spx?ar ticleID=jcim20110517
Original Experimental Research　实验论著
Antidiabetic and hypolipidemic activities of Dillenia indica
extract in diabetic rats
Sunil Kumar , Vipin Kumar , Om Prakash
Institute o f Pharmaceutical Sciences , Kurukshetra Unive rsity , Kurukshetra 136119 , Haryana , India
Objective:The present study was carried out to investigate the ant idiabet ic and hypolipidemic
activities of bioactive fraction(ethyl acetate fract ion)of Dillenia indica methanolic extract in
experimental diabet ic Wistar rats.
Methods:Type-1 diabetes was induced by single intraperitoneal injection of streptozotocin(60 mg/kg body weight), and type-2 diabetes was induced by single intraperitoneal injection
of st reptozotocin(60 mg/kgbody weight), 15 min after the intraperitoneal injection of 120 mg/kg
nicot inamide.The rats were treated by administering graded oral doses of isolated ethyl
acetate fraction of methanolic extract of D .indica (DIEE), 200 and 400 mg/kg body weight ,
respectively , for 21 d.The blood glucose level was estimatedat weekly intervals by glucometer.
Serum cholesterol , triglycerides and high-density lipoprotein cholesterol (HDL-C)were also
evaluated in normal and diabetic rats by an autoanalyzer.
Results:In both experimental models , daily oral treatment with DIEE for 21 d resulted in a
significant reduction in blood glucose (P <0.01), serum cholesterol (P <0.05)and
triglycerides(P <0.05)levels whereas HDL-C level was found to be increased(P <0.05)as
compared with the diabetic control group.
Conclusion:DIEE at 400 mg/kg body weight has prominent antidiabetic effect in experimental
type-1 and type-2 diabetes models in rats.It may therefore be used as an alternat ive remedy
for the treatment of diabetes mellitus.
Keywords:Dilleniaceae;plant extracts;diabetes mellitus , experimental;hypoglycemic agents;
antilipemic agents;rats
　Diabetes mellitus is a metabolic disorder resulting
from a defect in insul in secr etion , insulin act ion
or both.I nsulin deficiency in turn leads to chronic
hyperglycemia with disturbances of carbohydrates ,
f at and protein me tabol ism
[ 1] .Worldwide , total
number of people wi th diabetes is projected to
increase fr om 171 million in 2000 to 366 million in
2030
[ 2] .The pathogenesis of diabetes mellitus is
·570· 中西医结合学报 2011年 5月第 9卷第 5期　Journal of Chinese Integ rat ive Medicine , May 2011 , Vol.9 , No.5
managed by insul in and oral administr at ion of
hypoglycemic drugs such as sulfonylureas and
biguanides
[ 3] .Unf or tuna tely , having numbers of
side effects , they are not suitable for use during
pr egnancy
[ 4 , 5] .Plants are well-known in t radi-
tional herbal medicine f or their hypoglycemic
activit ies , and available l iter ature indicates th at
there are more than 800 plant species showing
hypoglycemic activity
[ 4] .Plants used in traditional
medicine to tre at diabetes mel litus represent a
valuable alternative for the control of this disease.
Therefore , plant materials are continuously scrutinized
and explored for their ef fect as hypoglycemic
agents.　Dillenia indica belongs to the family Dilleniaceae ,
commonly called Elephant tree.The fruit
shows laxat ive propert ies and is used for r el ieving
abdominal pain.The bark and leaves have astringent
effects
[ 6] .The juice of D .indica le aves , bark and
fruits are mixed and given or ally for the tre atment
of cancer and diarr hea
[ 7] .Frui t and leaf ext racts
of D .indica are reported to have ant ioxidant
activity
[ 8 , 9] .Centr al ner vous system depr essant[ 10]
and ant i-inf lammator y act ivity[ 11] in mice and
antimicrobial activity
[ 12]
were found from the alcoholic
ext ract of the leaves of D .indica .D .indica
extract r evealed significant cytotoxic act ivity
when tested by brine shrimp lethality bioassay and
showed antimicrobial activity against a wide range
of Gram-positive and Gram-nega tive bacter ia and
fungi
[ 9] .The seed extract showed hepatoprotective
effect
[ 13] .Earl ier phytochemical studies showed
that D .indica contains triterpenoids and flavonoids
such as lupeol , betulin , betulinaldehyde dillene tin ,
betulinic acid , β-sitosterol , st igmaster ol and
myricetin
[ 9, 14 , 15] .The fruits also showed significant
antileukemic activity in human
[ 16] .Tradit ion ally ,
the plant is also used in t reatment of diabetes
[ 17] .
The re is no previous research wor k available on
antidiabet ic use of the plant.So , the present
work was per formed to explore the antidiabetic
and hypolipidemic activit ies of isolated ethyl acetate
fract ion of meth anolic extr act of D .indica(DIEE).
1　Materials and methods
1.1 　Drugs and reagents　Streptozotocin (STZ ;
Lot Numbe r:000091422) was pur chased from
Sigma-Aldrich , I ndia and nicotinamide was purchased
from Hi-Media , India.Total cholesterol , high-density
lipoprotein cholesterol (HDL-C)and triacylglycerol
wer e assayed by an autoanalyzer (Erba Chem 7 ,
Mannheim , Germany)by using standard kits from
Erba Diagnostics Mannheim Gambh , Germany and
blood glucose level was measured by using a
glucose meter (Elegance CT-X10)of Conver gent
Technologies , Germany.All the reagents used in
this study were of analytical grade.
1.2　Experimental animals　Wistar rats of ei ther
sex , weighing about 150 to 250 g were used in the
study.Animals were maintained under standard
envir onmental condit ions i.e.ambient temper a-
ture of (22±2)℃ and a t 45% to 55% relative
humidity for 12 h , each of dark and ligh t cycle
and f ed with a standard pel let rats diet obtained
fr om Ashirwad Indust ries , Chandigarh , India , and
water was supplied ad l ibi tum .All the studies
were conducted in accordance with the Animal
Ethical Commit tee of the Kurukshetr a University ,
India.
1.3　Plant materials　D .indica leaves were collected
fr om the campus of Kurukshet ra Universi ty ,
Kur ukshetr a , India and were ident ified by Dr.
H .B.Singh , Scient ist F &Head , Raw Mater ial
Herbarium &Museum , National Institute of Science
Communication and Information Resources(NISCAIR),
New Delhi , India.A voucher specimen of the
plant is preserved in the her bar ium (NISCAIR/
RHMD/Consul t/-2009-10/1381/182/1).
1.4 　Extract preparation and fractionation 　The
collected leaves were washed with dist illed water
and dried in shade.The dried leaves were powdered
by using dry grinder and passed through a sieve.
The powdered mater ial was packed into Soxhlet
appara tus and extracted with methanol below 60 ℃.
The extract was evapor ated to dryness under
reduced pressure at 45 ℃ to yield sol id residues.
The sol id extract was suspended in water and
successively extr acted with hexane , ethyl acetate
and n-butanol in separ at ing funnel.Each f raction
was stored in air tight containers in a refr igerator
below 10 ℃ for subsequent expe riments.
1.5　Induction of type-1 diabetes model(T1DM):
insulin-dependent diabetes mell itus　Animals we re
made diabe tic by single intr aperi toneal injection
of STZ (60 mg/kg body we ight)dissolved in
0.1 mol/L citr ate buff er , pH 4.5.The blood
glucose level was checked before and 72 h after
STZ injection to conf irm the development of
diabetes.Only those animals which showed
hyperglycemia(blood glucose level>250 mg/dL)
were used in the exper iment.
1.6　Induction of type-2 diabetes model(T2DM):
non-insulin-dependent diabetes mellitus　T2DM was
induced in overnight fasted animals by a single
intraperitoneal injection of 60 mg/kg body weight
STZ , 15 min after the int raperi toneal inject ion of
120 mg/kg body weight nicotinamide.Hyperglycemia
was confirmed by the elevated blood glucose level
determined a t 72 h after and then on day 7 of the
inject ion.Only r ats conf irmed with pe rmanent
T2DM were used in the antidiabet ic study[ 18] .
1.7　Biochemical parameters　Blood samples we re
taken from the t ip of the tail of each rat of different
gr oups under mild ether anesthesia , and glucose
levels were determined by using blood glucose test
st rips with Elegance glucome ter (Frankenberg ,
Germany)at weekly intervals i.e.0 , 7 , 14 and
21 d af ter daily administr at ion of the extract
orally.Serum cholesterol , triacylglycerol and HDL-C
·571·中西医结合学报 2011年 5月第 9卷第 5期　J ou rnal of C hinese Integrative Medicine , May 2011 , Vol.9 , No.5
wer e also evaluated in normal and diabe tic r ats by
an autoanalyzer using Erba diagnostic ki ts
[ 19 , 20] .
1.8 　Experimental design 　 Overnight fasted
normoglycemic and diabet ic rats wer e divided into
seven gr oups with six rats e ach and tre ated or ally
daily for 21 d as follows.Group Ⅰ :normal
healthy control receiving only vehicle(Tween 80 ,
2% v/v);group Ⅱ:type-1 diabet ic contr ol
receiving only veh icle (Tween 80 , 2% v/v);
group Ⅲ:type-2 diabe tic control receiving only
vehicle (Tween 80 , 2% v/v);gr oup Ⅳ:type-1
diabet ic r ats were t reated with DIEE1(200 mg/kg
body weight);group Ⅴ:type-1 diabetic rats treated
with DIEE2(400 mg/kg body weight);group Ⅵ :
type-2 diabetic rats treated with DIEE1(200 mg/kg
body we ight);group Ⅶ :type-2 diabetic r ats
tr eated with DIEE2(400 mg/kg body weight).
1.9　Statistical analysis　All the data were presented
as x ±s x .The stat istical analysis involving two
groups was evaluated by means of one way analysis
of variance followed byDunnett-t multiple comparison
post-test which was used for statist ical comparison
between contr ol and various tre atment groups.
Sta tist ical signif icance was accepted at P <0.05.
2　Results
2.1　Effect on T1DM　STZ induced T1DM in rats
with fasting blood sugar level more than 250 mg/dL.
Daily or al administr ation of DIEE at 200 and
400 mg/kg significant ly(P <0.01)reduced blood
glucose level in diabe tic r ats(Table 1).
2.2　Effect on T2DM　STZ-nicot inamide induced
T2DM diabetes in r ats.Blood glucose level was
increased in diabetic control gr oup.Significant
reduct ion (P <0.01)in the blood glucose levels
was brough t about in diabet ic rats by daily admin-
ist rat ion of DIEE at 400 mg/kg(Table 1).
2.3　Effect on l ipid profile　Daily t reatment with
DIEE at 200 and 400 mg/kg for 21 d caused
signif icant improvement in the HDL-C level as
compared with the diabe tic control gr oups.Serum
cholesterol and triglyceride levels were decreased
signif icantly(P <0.01)by DIEE in T1DM and
T2DM rats(Table 2).
Table 1　Effect of DIEE on the blood glucose levels in diabetic rats (x±sx , mg/L)
G roup n
Blood g lucose level
Day 0 Day 7 Day 14 Day 21
Ⅰ :normal cont rol 6 1 102.3±56.0 1 082.3±48.0 1 125.0±34.0 1 154.7±39.0
Ⅱ:t ype-1 diabet ic cont rol 6 2 615.0±32.2 2 945.0±37.1 3 556.0±13.2 4 050.0±35.8
Ⅲ:t ype-2 diabet ic cont rol 6 2 783.0±35.2 3 044.9±42.5 3 753.1±24.2 4 152.0±39.7
Ⅳ:t ype-1 diabet ic+DIEE(200 mg/kg body w eight) 6 2 652.3±34.8 2 402.4±35.7 1 577.3±24.2＊ 1 288.4±22.9＊＊
Ⅴ:t ype-1 diabet ic+DIEE(400 mg/kg body w eight) 6 2 745.8±37.6 2 555.2±34.7＊ 1 523.8±34.6＊＊ 1 193.4±34.5＊＊
Ⅵ :t ype-2 diabet ic+DIEE(200 mg/kg body w eight) 6 2 892.3±35.7 2 632.5±34.5 1 974.3±46.8 1 385.8±34.5■
Ⅶ :t ype-2 diabet ic+DIEE(400 mg/kg body w eight) 6 2 853.5±32.5 2 685.7±24.2 1 422.5±26.9■■ 1 222.5±45.5■
　　＊P<0.05 , ＊＊P<0.01 , vs group Ⅱ;■P<0.05 , ■■P<0.01 , vs group Ⅲ.DIEE:i solated ethyl acetate f raction of meth anolic ext ract
of D.ind ica.
Table 2　Effect of DIEE on lipid prof ile in diabetic rats af ter 21 d of treatment (x±sx , mg/L)
G roup n Total cholesterol T riacylglycerol H igh-den sity lipoprotein-cholesterol
Ⅰ :normal cont rol 6 872.8±38.0 824.2±51.6 373.2±29.0
Ⅱ:t ype-1 diabet ic cont rol 6 2 547.3±76.0 1 505.2±47.1 282.3±22.0
Ⅲ:t ype-2 diabet ic cont rol 6 2 575.4±54.0 1 564.3±32.5 243.7±25.0
Ⅳ:t ype-1 diabet ic+DIEE(200 mg/kg body w eight) 6 1 185.8±45.0 1 048.7±25.4＊ 321.3±38.4＊
Ⅴ:t ype-1 diabet ic+DIEE(400 mg/kg body w eight) 6 1 132.4±34.0 984.7±32.5＊ 354.4±32.4＊＊
Ⅵ :t ype-2 diabet ic+DIEE(200 mg/kg body w eight) 6 1 274.4±34.0■ 1 072.4±44.6 303.4±39.0■
Ⅶ :t ype-2 diabet ic+DIEE(400 mg/kg body w eight) 6 1 174.3±49.0■ 1 023.0±22.6■ 344.4±43.0■
　　＊P<0.05 , ＊＊P<0.01 , vs group Ⅱ;■P<0.05 , ■■P<0.01 , vs group Ⅲ.DIEE:i solated ethyl acetate f raction of meth anolic ext ract
of D.ind ica.
3　Discussion
　STZ possesses diabetogenic proper ties mediated
by dest ruct ion of pancre atic β-cel ls and h as been
widely used to induce diabetes in experimen tal
animals
[ 21] .STZ-nicotinamide could induce type-2
diabetes in r ats.The present resear ch work was
carried out to evaluate antidiabet ic and hypolipi-
demic act ivi ties of DIEE in both T1DM and T2DM
rat models.DIEE brought significant reduction in
blood glucose level in both types of diabet ic rats.
Lipid abnormality accompanying with atherosclerosis
is the major cause of cardiovascular disease in
diabetes.I deal tre atment for diabetes , in addition
to glycemic control , should have a favorable effect
on lipid prof iles.Abnormal ities in l ipid prof ile
ar e found in about 40% of diabetics[ 22] .Untreated
diabetes is also associated with hypercholesterolemia
and hypert riglyceridemia
[ 23] .In the present work ,
DIEE reduced the total cholesterol and triglycerides
·572· 中西医结合学报 2011年 5月第 9卷第 5期　Journal of Chinese Integ rat ive Medicine , May 2011 , Vol.9 , No.5
levels as compared with the diabet ic contr ol
groups.There was also beneficial eff ect on HDL-
C.Th is hypol ipidemic effe ct may be due to inhi-
bit ion of fat ty acid synthesis
[ 24] .Therefore , DIEE
could be useful in managing both types of diabetes
as well as abnormal l ipid profile.The antidiabetic
activity of the plant may be due to the presence of
bioact ive const ituents such as t rite rpenoids and
flavonoids.
4　Conclusion
　The r esults indicate that the DIEE is effect ive
in decre asing the blood glucose level in diabetic
animals and also have beneficial effect on l ipid
profile.However , the nature of the active constituents
responsible for such an effe ct requires fur ther
invest igat ion.The possible mode of act ion of the
plant ext ract might be by potent iat ion of the insulin
effect by increasing the pancrea tic secr etion of
insulin from β-cells of islet of Langerh ans or its
release fr om the bound f orm or regenera tion of
the cells.
5　Competing interests
　The authors declare that they have no competing
interests.
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五桠果提取物对糖尿病大鼠的降血糖及降血脂作用
Sunil Kumar , Vipin Kumar , Om Prakash
Institute o f Pharmaceutical Sciences , Kurukshetra Unive rsity , Kurukshetra 136119 , Haryana , India
目的:使用实验性糖尿病Wistar大鼠模型验证五桠果(Dillenia indica)甲醇提取物的生物活性成分(乙酸乙
酯馏分)的降血糖及降血脂作用。
方法:通过腹腔内注射链脲佐菌素(60 mg/kg)建立大鼠 1型糖尿病模型 ,腹腔内注射烟酰胺(120 mg/kg)
15 min后腹腔内注射链脲佐菌素(60 mg/kg)建立大鼠 2型糖尿病模型 。在不同的实验模型中 ,用药组大鼠
均分别给予 200 mg/kg 和 400 mg/kg 的五桠果甲醇提取物的乙酸乙酯馏分连续 21 d。使用血糖仪每周 1
次测定各组大鼠血糖水平 。使用自动分析仪器分别测定各组大鼠血清总胆固醇 、三酰甘油及高密度脂蛋白
胆固醇水平。
结果:在两个实验模型中 ,与模型组比较 ,五桠果甲醇提取物的乙酸乙酯馏分均能显著降低模型大鼠的血糖
水平(P <0.01)以及血清总胆固醇(P<0.05)和三酰甘油(P <0.05)含量 ,升高高密度脂蛋白胆固醇水平
(P<0.05)。
结论:五桠果甲醇提取物的乙酸乙酯馏分在 400 mg/kg 剂量时 ,对于 1型及 2型糖尿病模型大鼠具有显著的
治疗作用 。因此这种提取物有望被研究开发成为治疗糖尿病的又一替代药物。
关键词:五桠果科;植物提取物;糖尿病 , 实验性;降血糖药;降血脂药;大鼠
2011全国时间生物医学学术会议征文通知
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　　会议时间　2011 年 9 月 15 ～ 18 日 , 会期 4 天。
　　会议地点　南宁天妃商务酒店(南宁市兴宁区明秀东路 238 号)。
　　会议内容　(1)学术交流:①现代时间生物医学研究方法与技术 、生物节律基因控制 、时间生理 、时间药理 、时间毒理 、时间
治疗 、时间护理 、时间营养 、睡眠 、时差及轮班不适应症的防治等相关研究;②中医时间医学与时间针灸 , 包括脏腑经络气血生
理节律 、疾病病理节律 、疾病时间治疗 、时间养生保健 、传统时间针灸等相关研究;(2)国内外时间生物学著名专家作有关时间
生物学研究进展学术报告;(3)时间生物医学专业委员会换届;(4)成立时间生物医学青年委员会。
　　征稿要求　论文全文(包括 400 字以内的中 、英文摘要 ,研究性论文请按目的 、方法 、结果 、结论四部分撰写), 以电子邮件
的附件方式(Word 文件)发送至会议联系人邮箱(联系人:赵子彦;电子信箱:ziy anzhao@163.com 或 ziy anzhao@sina.com)。
截稿日期:2011 年 7 月 31 日。详情参见中国时间生物医学网:http://w w w.chr ono-biolog y.ne t。
　　会务费　800元/人 ,学生 400 元/人 ,食宿及交通费用自理。
中国中西医结合学会
·574· 中西医结合学报 2011年 5月第 9卷第 5期　Journal of Chinese Integ rat ive Medicine , May 2011 , Vol.9 , No.5