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DOI:10. 3736 /jcim20120213
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Rahma tullah M , Hasan SK , Ali Z , Rahman S , Jahan R.
Antihyperg lycemic and antinociceptive activities of me thanolic
e xt ract o f Euphorbia thymi f olia L. w hole plants. J Chin
Integr Med . 2012;10(2):228-232.
Rahma tullah M , Hasan SK , Ali Z , Rahman S , Jahan R.
千根草甲醇提取物的抗高血糖及镇痛作用. 中西医结合
学报. 2012;10(2):228-232.
Received Octobe r 1 , 2011;accepted November 1 , 2011;
published online February 15 , 2012.
Full-tex t LinkOut at PubM ed. Journa l title in PubMed:
Zhong X i Y i J ie He X ue Bao.
基金项目:The study w as suppo rted by internal funding
from the U nive r sity o f Development A lte rna tive ,
Bangladesh
Correspondence:Mohammed Rahmatullah , PhD , Professo r;
Tel:+88-02-9136285;E-mail:rahamatm@hotmail. com
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O rig inal Expe rimental Research 实验论著
Antihyperglycemic and antinocicept ive activities of
methanolic extract of Euphorbia thymifolia L.
whole plants
Mohammed Rahmatullah , Sumaiyah Kanij Hasan , Zulfiquar Ali , Shahnaz Rahman , Rownak
Jahan
Depar tment o f Biotechno lo gy and Genetic Eng ineering , Faculty of Life Sciences , Univer sity o f Development
Alternativ e , Dhanmondi , Dhaka 1205 , Bang ladesh
OBJECTIVE:To study the antihyperglycemic and antinocicept ive act ivities of methanolic
extract of the whole plant of Euphorbia thymifolia L. , a plant used in folk medicine of Bangladesh
for treatment of diabetes and pain.
METHODS:Antihyperglycemic activity studies were conducted in glucose-loaded mice by oral
glucose tolerance tests. Mice were given various doses of the extract , followed by glucose(2 g/kg body weight), 1 h after administration of the extract. Serum glucose levels were
measured 2 h after glucose administ rat ion. Antinocicept ive activity studies were conducted in
intraperitoneally acetic acid-injected mice through measurement of reductions in abdominal
writhing t imes caused by acetic acid-induced gastric pain. Following a period of 1 h after oral
administration of various doses of the extract , all mice received intraperitoneal injection of 1%
acetic acid at a dose of 10 mL/kg body weight. To ensure bioavailability of acetic acid , a
period of 5 min was given to each animal following which period the number of writhings was
counted for 10 min.
RESULTS:The extract caused a significant dose-dependent reduction in serum glucose levels
in mice , when administered at doses of 50 , 100 , 200 and 400 mg/kgbody weight as compared
to the control animals (P <0. 05). The highest reduct ion of serum glucose (60. 5%) was
observed at a dose of 400 mg/kg. In comparison, a standard antihyperglycemic drug
glibenclamide , when administered at a dose of 10 mg/kg body weight , lowered serum glucose
levels by 48. 6 %. The extract also demonstrated a significant dose-dependent antinociceptive
activity compared to control animals (P <0. 05). At a dose of 400 mg/kg body weight , the
number of abdominal writhings was inhibited by 40. 9% as compared to 49. 0% inhibition
obtained with a standard antinociceptive drug aspirin, administered at a dose of 200 mg/kg
228 中西医结合学报 2012年 2月第 10卷第 2期 Journal of Chinese Integrat ive Medicine , February 2012 , Vol. 10 , No. 2
body weight.
CONCLUSION:The significant ant ihyperglycemic and antinocicept ive act ivities demonstrated
by the extract validate the use of E . thymifolia in folk medicine of Bangladesh for t reatment of
diabetes and pain , and merit further scientific studies leading to discovery of efficacious drugs.
KEYWORDS:Euphorbia ;plant extracts;hypoglycemic agents;analgesics;mice
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Zhu Y , Liu JR, Zhang W. An overv iew o f medicinal plant re source fo r decreasing blood gluco se. J Chin Integ r Med. 2004;
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Euphorbia thymifolia L牣牗Euphorbiaceae牘 is
found in wild and neglected areas of Bangladesh牣
The plant is used in folk medicinal system of the
country for tr eatment of hypertension牞 hear t
diseases牞helminthiasis and leucorrhea牣I t is also
occasionally used f or t reatment of diabe tes and
pain牣The plant is bit ter in taste and used in t radi-
tional I ndian medicine for const ipation牞helmin-
thiasis牞ringworm牞skin diseases and leprosy犤1犦牣
The leaves and seeds are used for bowel distur b-
ances in children and ar e considered laxative
犤2犦牣
Antibacter ial act ivity h as been repor ted for the
plant
犤3犦牣 Flavonoids extracted from the aerial
parts of the plant repor tedly showed high cytotoxicity
on human epidermoid cancer cells and moder ate
antiviral act ivities
犤4犦牣Antioxidant and antiviral
activit ies have also been reported for methanol牞
chloroform牞ethyl acetate牞n-butanol and aqueous
extracts of the plant as well as several isolated pure
compounds like r ugosin B牞3-O-galloyl-4牞6-牗S牘-
HHDP-D-glucose and 1牞3牞4牞6-te tr a-O-galloyl-K-
beta-D-glucose犤5犦牣Ethyl acetate fr action of the
plant repor tedly exh ibited anti-herpes simplex
virus-2 activity in vi tro犤6犦牣 Diabetes is a debilitating disease affect ing millions
of people worldwide which is projected to rise
because of changes in lifestyle牣Pain牞 resul ting
from var ious causes牞also affects millions of people
worldwide on a da ily basis牣Modern allopathic
medicine has no known cure for diabetes牣Although
there are effect ive medicines f or pain l ike aspirin
or par acetamol牞 these medicines can cause side
effects like gastric ulceration and hepatic damages牣
The folk medicinal pr acti tioners of Bangladesh
牗Kavirajes牘claim to have effect ive tre atments for
both diabetes and pain without resul tan t toxicity
as occuring with allopathic pain killers牣We had
been scr eening the var ious anti-diabet ic and ant in-
ocicept ive medicinal plants used by the Kavir ajes
for some t ime to determine the ir actual efficacies
for the purposes tha t they are used for
犤7-10犦牣The
object ive of the present study was to evaluate the
ant ihyperglycemic and antinocicept ive act ivit ies
of me thanol ic extr act of E牣thymifol ia whole
plants牣
1 Materials and methods
1牣1 Plant materials and extraction Whole plants
of E牣thymi fol ia wer e collected f rom Dhaka
distr ict牞Bangladesh in November 2010牣The plant
was taxonomically identif ied by the Bangladesh
Nat ional Her bar ium at Dhaka牗accession number牶
35394牘牣The whole plants were air-dried in the
shade for 120 h牞grounded into a fine powder牞
and were extr acted with methanol at a r atio of
1∶6牣25牗weight牤volume牘牣The init ial weight of
dried powder used for extr action was 80 g牷the
final we ight of the me thanol extr act obtained
from the dried powder was 5牣96 g牗yield =7牣45%牘牣
1牣2 Chemicals and drugs Glibenclamide牞aspirin
and glucose wer e obtained fr om Square Pharma-
ceut icals Ltd牣牞Bangladesh牣All other chemicals
were of analytical grade牣
1牣3 Animals I n the pr esent study牞Swiss albino
mice牗male牘牞which weighed between 17 to 22 g
were used牣The animals were obtained from the
International Center for Diarrheal Disease Research牞
Bangladesh牣All animals were kept under ambient
tempera tur e with 12 h light followed by 12 h
dark牣The animals were accl ima tized for 3 d prior
to actual exper iments牣The study was conducted
following approval by the I nstitut ional Animal
Ethical Committee of the University of Development
Alternat ive牞Dhaka牞Bangladesh牣
229 中西医结合学报 2012年 2月第 10卷第 2期 J ou rnal of C hinese Integrative Medicine , Feb ruary 2012 , Vol. 10 , No. 2
1牣4 Antihyperglycemic activity Glucose tolerance
pr operty of the me thanol extr act of the whole
plant was de termined as per the procedure
pr eviously described by Joy et al
犤11犦
with minor
modif ications牣I n brief牞fasted mice were grouped
into six groups with six mice in each group牣The
various groups received diff erent trea tments牣
Group 1 received vehicle牗1 %Tween 80 in water牞
10 mL牤kg body weight牘 and served as the normal
control牷group 2 received standard drug牗glibenclamide牞
10 mg牤kg body weight牘牷groups 3 to 6 received
methanol ext ract of E牣thymi fol ia whole plant at
doses of 50牞 100牞 200 and 400 mg牤kg body
weight牞 respect ively牣 Doses were prepared by
thoroughly suspending methanol ext ract in 1 %
Tween 80 in water牣Each mouse was we ighed and
dose-adjusted accordingly prior to administ rat ion
of the vehicle牞standar d dr ug牞and test samples牣
All substances were orally administered牣Following a
per iod of 1 h牞all mice were orally administer ed
2 g牤kg glucose牣Blood samples wer e collected 2 h
after the glucose administ rat ion by puncturing
heart of the mice牣Ser um glucose levels were
measur ed by glucose oxidase method
犤12犦牣
1牣5 Antinociceptive activity Antinocicept ive
activity of the meth anol ext ract of E牣thymi folia
whole plant was examined by using pr eviously
described procedures
犤13犦牣Briefly牞mice were divided
into seven groups with six mice in each group牣
Group 1 ser ved as the normal contr ol and was
administe red vehicle only牷gr oups 2 and 3 were
or al ly administered standard ant inociceptive dr ug
aspi rin at doses of 200 and 400 mg牤kg body
weight牞respectively牣Groups 4 to 7 were administered
methanolic extr act of E牣thymi folia whole plant
at doses of 50牞100牞200 and 400 mg牤kg body
weight牞respect ively牣Following a period of 1 h
after or al administ rat ion of the standard drug or
extracts牞all mice received intraperitoneal injection
of 1% acetic acid at a dose of 10 mL牤kg body
weight牣To ensure bioavailability of acet ic acid牞a
per iod of 5 min was given to each animal following
which period the number of writhings was counted
for 10 min牣
1牣6 Statistical analysis Results were expressed
as mean ± standard er ror of mean牣Stat istical
analysis was performed using one-way analysis of
variance by a stat ist ical package SPSS version
15牣0牣To determine the significant diff erences
between different gr oups牞LSD-t test was done牣
P <0牣05 impl ied sta tist ical significance牣
2 Results
2牣1 Antihyperglycemic activity In oral glucose
toler ance test conducted with glucose-loaded mice牞
the methanolic ext ract demonstr ated a significant
dose-dependent antihyperglycemic activity牣Following
administr ation of the standard ant ihyperglycemic
drug gl ibenclamide牞serum glucose concentr ation
was significantly r educed by 48牣6 %牗P <0牣05牘牣
All tr eatments with var ious doses of the extract
led to signif icant reductions in ser um glucose
levels as compared to the contr ol gr oup 牗P <
0牣05牘牣The extract牞when administered a t doses
of 50牞100牞200 and 400 mg牤kg body weight
reduced serum glucose concentrat ions by 30牣8%牞
42牣0%牞47牣6%牞and 60牣5%牞respect ively牣Even
a t a dose of 200 mg牤kg牞the result can be favorably
compared with that of the glibenclamide group牣
The results are shown in Table 1牣
2牣2 Antinociceptive activity The extr act also
showed dose-dependent reductions in the number
of wr ithings in mice induced by acetic acid int rap-
eritoneal administ rat ion in the ant inociceptive
act ivity test牣The results were significant for all doses
of ext ract-administered mice牞when compared to
the control gr oup牗P <0牣05牘牣At the highest dose
of the extr act tested牗400 mg牤kg牘牞the number of
writhings was reduced by 40牣9%牣In contr ast牞a
standard antinocicept ive drug aspirin reduced the
number of writhings by 49牣0 % and 67牣3%牞
respectively牞when administered a t doses of 200
and 400 mg牤kg body weight牣 The results are
shown in Table 2牣
3 Discussion
Antihyperglycemic and ant inociceptive activities of
the extr act were measured in two separate groups
of animals牣 This is the standard pr ocedure as
reported in scient ific li tera tur e牞for the two pro-
cedures for measurement of the two activit ies are
different牣I f using the same set of experiment on
the same group of animals then there could be
complications in the results obtained牣For instance牞
the pain cr eated in mice by int raperitoneal admin-
istr at ion of acetic acid and the consequent writhing
t imes may aff ect the levels of blood glucose in the
experimental animals which may be erroneously
Table 1 Antihyperglycemic activity evaluation of the methanolic extract of whole plant of Euphorbia thymifolia in mice
牗Mean±standa rd e rro r o f mean牘
Group n Serum glucose level牗mg牤L牘 Low ering of s erum glu cose level牗%牘
C on t rol牗10 mL牤kg 1% Tw een 80牞p牣o牣牘 6 1 288牣3±42牣3 -
Glib enclamide牗10 mg牤kg牞p牣o牣牘 6 662牣2±79牣2* 48牣6
Euphorbia thymi fol ia ext ract牗50 mg牤k g牞p牣o牣牘 6 891牣9±79牣2*△ 30牣8
Euphorbia thymi fol ia ext ract牗100 mg牤k g牞p牣o牣牘 6 747牣7±101牣3*△ 42牣0
Euphorbia thymi fol ia ext ract牗200 mg牤k g牞p牣o牣牘 6 675牣7±86牣0* 47牣6
Euphorbia thymi fol ia ext ract牗400 mg牤k g牞p牣o牣牘 6 509牣0±47牣5* 60牣5
*P<0牣05牞vs con t rol group牷△P <0牣05牞vs glibenclamide group牣p牣o牣牶per oral牣
230 中西医结合学报 2012年 2月第 10卷第 2期 Journal of Chinese Integrat ive Medicine , February 2012 , Vol. 10 , No. 2
Table 2 Antinociceptive activity evaluation of the methanolic extract of whole plant of Eupohorbia thymif olia in mice
牗Mean±standa rd e rro r o f mean牘
Group n Number of w ri things Inhibi tion牗%牘
C on t rol牗10 mL牤kg 1% Tw een 80牞p牣o牣牘 6 8牣17±0牣79 -
Aspirin牗200 m g牤kg牞p牣o牣牘 6 4牣17±0牣65* 49牣0
Aspirin牗400 m g牤kg牞p牣o牣牘 6 2牣67±0牣88* 67牣3
Euphorbia thymi fol ia ext ract牗50 mg牤k g牞p牣o牣牘 6 5牣50±1牣38*▲ 32牣7
Euphorbia thymi fol ia ext ract牗100 mg牤k g牞p牣o牣牘 6 5牣33±1牣05*▲ 34牣8
Euphorbia thymi fol ia ext ract牗200 mg牤k g牞p牣o牣牘 6 5牣00±0牣37* 38牣8
Euphorbia thymi fol ia ext ract牗400 mg牤k g牞p牣o牣牘 6 4牣83±0牣40* 40牣9
*P<0牣05牞vs con t rol group牷▲P <0牣05牞vs aspi rin group牗400 mg牤kg牘牣p牣o牣牶per oral牣
at tr ibuted to the extract牣However牞the extr act
had both antihyper glycemic and antinocicept ive
activit ies牗as separately determined牘which can be
ut ilized in tr eatment of pain in diabetic animals as
well as used separately for treatment of diabetes or
pain牣Notably牞this applies to the extract only牣If
antihyper glycemic and ant inociceptive activities
are due to different phytochemical const ituents of
the extr act which is possible牞then administration
of isolated responsible phytochemicals will demon-
st ra te only one type of act ivity牣 The obser ved lowering of blood glucose level by
the extract may be achieved through various individuals
or a combina tion of mechanisms牣The extr act may
have potent iated pancreat ic secret ion of insulin牞
increased glucose uptake from serum牞or decreased
glucose absorpt ion from gut
犤14-16犦牣 The potent
antihyperglycemic activity exhibited by the extract
mer its f urther studies for isolat ion of r esponsible
phytochemical components and such studies are
now being actively pursued in our labor atory牣 Acet ic acid-induced writhing test can detect
both centr al an d per ipher al analgesia
犤1 3犦牣In tr a-
pe ri toneal administ rat ion of acetic acid le ads to
occurrence of pain牞 which is caused mainly
thr ough increased pr oduct ion of prostacycl in and
pr ostaglandin E leading to excitation of the A-
del ta nerve fibers and so causing sensation of
pain
犤17犦牣Ther efor e牞any agent that lowers the
number of abdominal const rict ions wil l demon-
st ra te analgesia by inhibi tion of prostaglandin
synthesis牣I t is under investigation whether the
extract contains components which can inhibit the
synthesis of prostaglandins and so decr ease the
number of wr ithings induced by ace tic acid in
mice牣 Taken together牞both antihyperglycemic as well
as antinociceptive effects observed in E牣thymi folia
extract validate the folk medicinal uses of this
plant f or trea tment of diabetes and pain牣Many
allopa thic drugs like aspirin牞atropine牞ephedrine牞
digoxin牞morphine牞quinine牞reserpine and tubo-
curarine have been discover ed thr ough obser ving
indigenous medicinal pr actices
犤18犦牣I t is expected
tha t screening of folk medicinal plants of Bangla-
desh and scient ific validat ion of their relevant
pharmacological activit ies can open up new avenues
for discover ing more eff icacious drugs and at the
same t ime spur the conservation efforts of the
plant species because many of these medicinal
plants are r apidly becoming endangered thr ough
over-exploi ta tion牞spread of human habitat牞and
destr uction of forest areas牣
4 Financial support
The study was supported by an internal funding
fr om the Universi ty of Development Al terna tive牞
Bangladesh牣
5 Competing interests
The authors declare that they have no competing
interests牣
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千根草甲醇提取物的抗高血糖及镇痛作用
Mohammed Rahmatullah , Sumaiyah Kanij Hasan , Zulfiquar Ali , Shahnaz Rahman , Rownak Jahan
Depar tment of Biotechno log y and Genetic Eng ineering , Faculty o f Life Sciences , University of Development A lte rna tive ,
Dhanmondi , Dhaka 1205 , Bang ladesh
目的:本研究旨在证实孟加拉国传统药用植物千根草甲醇提取物的抗高血糖及镇痛作用 。
方法:实验分为独立的两个部分 。在研究千根草甲醇提取物抗高血糖作用的实验中 , 36只小鼠被分为 6组 ,
每组 6只 ,分别给予口服不同剂量的千根草甲醇提取物 、降血糖药格列本脲及 10 mL /kg 1% Tw een 80溶
剂 ,1 h后 ,予各组小鼠口服2 g /kg 葡萄糖 , 2 h后测取所有小鼠的血糖水平并进行比对。为观察千根草甲醇
提取物的镇痛作用 ,另外 42只小鼠被分为 7组 ,每组 6只 ,分别给予各组小鼠口服 10 mL /kg 1% Tween 80
溶剂 、不同剂量千根草甲醇提取物 ,或 200 和 400 mg /kg 镇痛药阿司匹林 1 h 后 ,向所有小鼠腹腔内注射
10 mL /kg 1%醋酸诱发扭体反应。诱发扭体反应 5 min后 ,每只小鼠观察 10 min ,记录扭体次数。
结果:分别口服 50 、100 、200和 400 mg /kg 千根草甲醇提取物的 4组小鼠与对照组相比血糖水平显著降低
(P <0. 05)。当小鼠口服 400 mg /kg 千根草甲醇提取物时 ,血糖水平下降最多。口服 10 mg /kg 格列本脲的
小鼠血糖水平下降 48. 6%。与对照组相比 ,甲醇提取物同样被证实具有显著的镇痛作用(P <0. 05)。当服
用剂量在 400 mg /kg 时 ,小鼠扭体次数抑制率达 40. 9%,而口服 200 mg /kg 镇痛药阿司匹林时 ,小鼠扭体次
数抑制率为 49. 0 %。
结论:孟加拉国民间草药千根草在治疗糖尿病与疼痛方面有显著作用 ,其甲醇提取物抗高血糖及镇痛的作用
值得进一步研究 ,以便开发出以此植物为基础的新型化合物与有效药 。
关键词:大戟属;植物提取物;降血糖药;镇痛药;小鼠
232 中西医结合学报 2012年 2月第 10卷第 2期 Journal of Chinese Integrat ive Medicine , February 2012 , Vol. 10 , No. 2