全 文 ：Open Access开放获取 Related Articles推荐阅读
DOI:10.3736/ jcim20110608
http:// ww w.jcimjournal.com
Ali MK , Ashraf A , Biswas NN , Karmakar UK ,
Af roz S.Antinociceptive , anti-inflamma to ry
and antidiar rheal activities of ethano lic calyx
extract of Hibiscus sabdari ff a Linn.(Malvaceae)
in mice.J Chin Integr Med .2011;9(6):
626-631.
Ali MK , Ashraf A , Bisw as NN , Karmaka r
UK , Afro z S.玫瑰茄花萼乙醇提取物的抗
痛觉敏感 、抗炎及止泻作用的实验研究.中
西医结合学报.2011;9(6):626-631.
Received February 8 , 2011;accepted March
2 , 2011;published online June 15 , 2011.
Full-tex t LinkOut a t PubMed.Journal title in
PubMed:Zhong X i Y i J ie He X ue Bao.
Correspondence:Md.Khadem Ali;Tel:+88-
01914-827647;E-mail:khadem bge05@yahoo.
com
Zaringhalam J , Akbari A , Tekieh E , Manaheji H , Rezazadeh S.桑托利纳
蓍降低弗氏佐剂致炎症大鼠的血清白细胞介素 6 水平及痛觉敏感程度.
中西医结合学报.2010;8(12):1180-1189.
Zaringhalam J , Akba ri A , Tekieh E , Manaheji H , Rezazadeh S.Achi llea
santolina r educe s serum interlukin-6 level and hyper alg esia during complete
F reund s adjuvant-induced inflammation in male Wistar rats.J Chin Integr
Med.2010;8(12):1180-1189.
Full text available at http:// www .jcimjournal.com/ FullText 2.aspx?articleID
=jcim20101211
Manaheji H , Jafa ri S , Zaringhalam J , Rezazadeh S , Taghizadfarid R.辣木
叶或根的甲醇提取物对弗氏佐剂致关节炎大鼠模型的止痛作用.中西医
结合学报.2011;9(2):216-222.
Manaheji H , Jafari S , Zaringhalam J , Rezazadeh S , Taghizadfarid R.
Analgesic effects of methano lic ex tracts of the leaf or r oo t o f Moringa
olei f era on complete F reunds adjuvant-induced ar thritis in rats.J Chin
Integr Med .2011;9(2):216-222.
Full text available at http:// www .jcimjournal.com/ FullText 2.aspx?articleID
=jcim20110216
More related articles at http:// ww w.jcimjournal.com/ FullTex t 2.aspx?articleID
=jcim20110608
Original Experimental Research 实验论著　
Antinociceptive , anti-inflammatory and antidiarrheal
act ivit ies of ethanolic calyx extract of Hibiscus
sabdariffa Linn.(Malvaceae)in mice
Md.Khadem Ali1 , Ayesha Ashraf1 , Nripendra Nath Biswas2 , Utpal Kumar Karmakar2 ,
ShamimaAfroz1
1.Bio technolog y and Genetic Enginee ring Discipline , Life Science Schoo l , Khulna Univer sity , Khulna-9208 ,
Bangladesh
2.Pharmacy Discipline , Life Science Schoo l , Khulna Univer sity , Khulna-9208 , Bang ladesh
Objective:To evaluate the antinociceptive , anti-inflammatory and antidiarrheal act ivities of the
ethanolic calyx extract of Hibiscus sabdariffa Linn.in mice.
Methods:In the present study , the dried calyxes of H .sabdariffa were subjected to extraction
with 95 %ethanol and the extract was used to investigate the possible activit ies.Ant inocicep-
t ive activity of the extract was evaluated by using the acetic acid-induced writhing test.The
anti-inflammatory effect of the extract was tested by using the xylene-induced ear edemamodel
mice.Castor oil-induced diarrheal model mice were used to evaluate the antidiarrheal act ivity
of the extract.
Results:In acet ic acid-induced writhing test , the extract produced inhibited writhing in mice
siginificantly compared with the blank control (P <0.01).The extract showed significant
inhibition of ear edema formation in xylene-induced ear edema model mice in a dose-related
manner compared with the blank control (P <0.01).The extract demonstrated a significant
antidiarrheal act ivity against castor oil-induced diarrheal in mice in which it decreased the
frequency of defecation and increased the mean latent period at the doses of 250 and 500 mg/kg
body weight (P <0.01).
Conclusion:The above mentioned findings indicate that the calyx extract of H.sabdariffa
possesses significant antinociceptive , anti-inflammatory and antidiarrheal activit ies that support
its uses in t raditional medicine.
Keywords:Hibiscus ;plant extracts;analgesics;anti-inflammatory agents;antidiarrheals;mice
·626· 中西医结合学报 2011年 6月第 9卷第 6期　Jou rnal of Chines e Integrative M edicine , June 2011 , Vol.9 , No.6
　Hibiscuss sabdarif fa is a medicinal herb of
Malvaceae family.I t is available in all parts of
the world and cul tivated for i ts leaf , fleshy calyx ,
seed and fibre.The dried red calyx of this plant is
commonly known as rosella in Aust ralia , mesta in
I ndian subcontinent , chin baung in Myanmar ,
krajeab in Thailand , zobo in Nigeria , karkade in
Egypt , Saudi Ar abia and Sudan and sorrel in
Lat in America
[ 1] .I t contains anthocyanins , β-
carotene , ascorbic acid , protocatechuric acid ,
mucilage , gossypetin , hibiscine cholorideare , calcium
citrate and cyanogenic glycoside
[ 1, 2] .H .sabdarif fa
has been tr adionally used for high blood pr essure ,
liver diseases , fever , ulcers , abscesses , anemia , etc[ 3] .
I t has also been reported to be diuretic , astringent ,
antisept ic , aphrodisiac , cholagogue , demulcen t ,
digestive , emollient , purgative , hypocholesterolemic ,
mucolytic , laxative , refrigerant , sedative , stomachic
and tonic
[ 4 , 5] .　Previous scient ific studies have reported th at
different parts of H .sabdari ffa possess antioxidant
and hypotensive
[ 6] , anxiolytic and sedat ive[ 7] ,
antipyret ic and ant iatherosclerot ic
[ 8] , antihyperlip-
idemic , hepatoprotective , antibacterial , antioxidant ,
anticancer , ant iclastogenic[ 1] , toxicity and immu-
nomodulator y
[ 9] , ant ihypertensive[ 10] , antimuta-
genic
[ 11] , ant ispasmodic[ 12] , cytotoxicity and ant i-
bacterial
[ 13]
effects .But scient ific invest igat ions
on antinocicept ive , anti-inflammatory and antidi-
arrheal activit ies of calyx extr act of this plant
have not ye t been performed which may suppor t
its uses in traditional medicine.The present study
was therefore undertaken to evaluate possible ant ino-
cicept ive , anti-inflammatory and ant idiarrheal
activit ies of ethanolic calyx extract of H .sabdarif fa
in mice.
1　Materials and methods
1.1 　Plant material collection and identification 　
The calyx of the plant was col lected from Shyam-
nagor unde r the district of Satkhira of Bangladesh
in December 2009 at day t ime.During the collec-
tion pr ocess , the calyxes were not washed or
cle aned by wa ter due to ch ance of hydrolysis ,
oxida tion and other types of chemical degrada-
tion.The soils and dusts th at were attached to the
calyxes were removed by hand shaking.During
the colle ct ion process , any type of adulterat ion
was st rongly prohibited.The plant was identif ied
by the experts of Bangladesh Nat ional Herbarium ,
Mir pur , Dhaka , Bangladesh and tagged with the
accession No.33858.The voucher specimen is
deposited in the Biotechnology and Genet ic Engi-
neering D iscipline , Khulna University , Khulna ,
Bangladesh.
1.2 　Plant extract preparation　Collected calyxes
wer e separ ated from undesirable plant parts and
ma terials.The calyxes were dr ied by air dr ying
for 10 d.Af ter drying , the calyxes were ground
into coarse powder with the help of a grinder.
The plant powder was stored in an air tight vessel
and kept in a cool and dry place.A glass jar with
plast ic cover was taken and washed thoroughly
and 100 g of the dried powder was taken into the
jar .95% ethanol (500 mL)was poured into the
jar up to 2.54 cm height above the sample sur face
as it can suff icient ly cover the sample sur face.
The plastic cove r with an aluminum foil was
closed pr operly to resist the entrance of air into
the jar and to avoid extract ion of chemicals in
con tact with the solven t. This pr ocess was
performed for 10 d.The jar was shaken sever al
t imes during the pr ocess to get better extr action.
Af ter the extract ion pr ocess the plant extr act was
fil tered by a piece of clean and whi te cot ton
mate rial twice.Then it was fil tered thr ough
Wh atman filter paper.The f il tr ate was col lected
in a beaker and evaporated under ceiling f an unt il
dried.It rendered a gummy concentrate of deep
red color .Finally , the extract was stored a t 4 ℃
until use.
1.3 　Experimental animals 　Young Swiss-albino
mice aged 4 to 5 weeks , average body weight 20
to 25 g wer e used for the experiment.The mice
were purchased fr om the animal house of the
Internat ional Center for Diarrhoeal Disease
Research , Bangladesh (ICDDR , B).They we re
kept in a standar d envir onmental condit ion for
one week in the animal house of the Biotechnology
and Genet ic Engineering Discipl ine , Khulna Uni-
versity , Bangladesh for adaptation.The animals
were provided with standard labora tory condi-
t ions (relat ive humidity 55% to 60%, room tem-
perature (25±2)℃ and a 12 h light/dark cycle).
All the experiments wer e conducted in an isolated
and noiseless condition.This study was conducted
accor ding to the guidelines of Inst itut ional Animal
Ethics Commit tee
[ 14] .
1.4　Chemicals and Drugs　Glacial acet ic acid and
xylene were purchased from Sigma chemicals ,
USA.The standard drugs diclofenac sodium and
lope ramide wer e collected fr om Square Pharma-
ceut icals Ltd., Bangladesh.
1.5 　Phytochemical screening 　Test of different
chemical groups pr esenting in the extract r epr e-
sents the preliminary phytochemical studies.Pr e-
liminary phytochemical analysis of the ethanolic
calyx extr act of H .sabdarif fa was carr ied out by
using the standar d procedure
[ 15 , 16] .
1.6　Antinociceptive activity　The ant inociceptive
act ivi ty of the plant ext ract was evaluated by
using the acet ic acid-induced wr ithing test in mice
following Ahmed et al
[ 17] .Mice were r andomly
selected and divided into 4 groups of 5 mice in
each group.Mice of gr oup Ⅰ served as the blank
con trol and received 1% Tween-80 orally at the
dose of 10 mL/kg body weight;mice of group Ⅱ
served as the posi tive control and received
diclofenac sodium orally at the dose of 25 mg/kg
body weight;mice of group Ⅲ and group Ⅳ as
test groups r eceived eth anolic calyx extr act of H .
sabdarif fa at doses of 250 and 500 mg/kg body
·627·中西医结合学报 2011年 6月第 9卷第 6期　J ou rnal of C hinese Integrative Medicine , Ju ne 2011 , Vol.9 , No.6
weight , respect ively.The rat ionale of choosing
the dosage is accor ding to crude drugs that had
been tr adi tionally used more or less at these doses
in humans f or curing var ious diseases.A 30 min
inter val was given to ensure proper absor ption of
the administered substances.Then the writhing-
inducing chemical , acet ic acid solution (0.7%,
10 mL/kg)was administered intr aper itoneally to
each mouse.An interval of 5 min was given for
absorpt ion of acet ic acid and the number of
wr ithing was counted for 15 min.The mice did
not always per form f ull wr ithing.The incomplete
wr ithing was taken as a hal f-wri thing , so two
half-wr ithings were taken as one full writhing.
This is why total writhing was halved to conver t
all wr ithing to full writhing or real wr ithing.
1.7 　Anti-inflammatory activity　Xylene-induced
ear edema model mice were used to assess the
anti-inflammatory activity of the plant extr act
following the method descr ibed by Dev et al
[ 18] .
Experimental mice were divided as the previous
test:administered vehicle (1% Tween 80 in
water , 10 mL/kg body weight);standard dr ug(diclofenac sodium , 10 mg/kg body weight)and
two diffe rent doses of plant extr act (250 and
500 mg/kg body weight).One hour after admin-
istr ation of the above dr ugs , 0.01 mL of xylene
was injected to the anter ior and poster ior surf aces
of the right ear of each mouse.One hour af ter
xylene injection , mice were sacri ficed and both
tr eated and untreated ears were cut down by using
a 7 mm diameter cor k borer as circular sect ions
and weighed.The weight difference between
untreated and tr eated ear sect ions was calculated.
1.8 　Antidiarrheal activity 　Castor oil-induced
diarrheal model mice were used to evaluate possible
antidiar rheal activity of the extract f ollowing the
method descr ibed by Chat ter jee
[ 19] .At first , all
the mice were screened afte r oral administ rat ion
of 0.5 mL of castor oil and only those showing
diarrheal were selected and randomly divided into
4 gr oups of 5 mice in each group.Group Ⅰ or the
contr ol r eceived only dist il led water con taining
1%Tween-80 a t a dose of 10 mg/kg body weigh t;
group Ⅱ or the posit ive control rece ived standar d
antimotil ity drug loperamide at a dose of 50 mg/kg
body we ight as or al suspension.The two test
groups were t reated with suspension of calyx
extract of H .sabdari ffa at or al doses of 250 and
500 mg/kg body weight.The mice were fed with
the samples 1 h prior to the or al administ rat ion of
castor oil.Mice of each group were placed in
separa te cages with adsorbent paper beneath and
examined for the presence of diarrhoea every hour in
the 4-hour study after the castor oil administr at ion.
The present invest iga tion was done for 4 h for
studying the delay of the onse t of diarrhea episode
or decrease of the frequency of defecation.Number
of stools or any fluid material that stained the
adsor bent paper were counted at each successive
hour during the 4-hour period and were noted for
each mouse.The latent per iod of each mouse was
also counted.
1.9　Statistical analysis　The da ta wer e presented
as x ±s x .Results were analysed by one-way
analysis of variance (ANOVA) followed by
Dunnet ts t test for mul tiple comparisons.The
signif icant diff erence was considered at P <0.05 .
2　Results
2.1　Phytochemical screening　Pr el iminar y phyto-
chemical screening of the ethanol ic calyx extract
of H .sabdari ffa revealed the presence of reducing
sugars , tannins , flavonoids , glycosides , alkaloids ,
saponins , and ster oids .
2.2 　Antinociceptive activity 　In the ace tic acid-
induced writhing test , the ethanol ic calyx extract
of H .sabdariffa showed significant dose-dependent
wri thing inhibit ion compared with the control
gr oup.Maximum wr ithing inhibition was 66.85%
a t the dose of 500 mg/kg body weight , which is
comparable to diclofenac sodium 78.45% at the
dose of 25 mg/kg body weight (Table 1).
2.3 　 Anti-inflammatory activity 　 In xylene-
induced ear edema model mice , the ethanolic
calyx extract of H .sabdarif fa produced significant
inhibit ion on ear edema formation in a dose-related
manner compar ed with the blank control group.
I t caused 18 .00% and 27.50% inhibit ion of e ar
edema forma tion at the doses of 250 and 500 mg/kg
body we ight , respect ively(Table 2).
2.4　Antidiarrheal activity　The effects of ethanolic
calyx extr act of H .sabdari ffa on the latent per iod
of castor oil-induced diarr heal model mice are
presented in Table 3.The extr act produced a
signif icant incre ase in the latent period in com-
parison with the blank control a t doses of 250 and
500 mg/kg body weight (P <0.01).I t also
signif icantly reduced the total number of faeces as
well as of diarr hoeic f aeces in a dose-dependent
manner compared with the blank control (P <0 .01)(Table 4).
Table 1　Effects of antinociceptive activity of ethanolic calyx extract of Hibiscus sabdarif fa in mice
Group n Number of w rithing (x±s x) Inhibi ti on
rate(%)
Blank con t rol(AA (10 m L/kg , i.p.)+vehicle(10 m L/kg , p.o.)) 5 36.20±2.03 00.00
Posi tive cont rol(AA (10 m L/kg , i.p.)+diclofenac sodium (25 mg/kg , p.o.)) 5 7.80±1.11＊＊ 78.45
Test 1 (AA (10 m L/k g , i.p.)+plant ex tract (250 mg/k g , p.o.)) 5 18.00±1.45＊＊ 50.27
Test 2 (AA (10 m L/k g , i.p.)+plant ex tract (500 mg/k g , p.o.)) 5 12.00±0.79＊＊ 66.85
　　＊＊P<0.01 , vs b lan k cont rol g rou p.AA:acet ic acid;i.p.:in t raperi toneally;p.o.:per oral.
·628· 中西医结合学报 2011年 6月第 9卷第 6期　Jou rnal of Chines e Integrative M edicine , June 2011 , Vol.9 , No.6
Table 2　Anti- inflammatory effect of ethanolic calyx extract of Hibiscus sabdari f fa on xylene-induced ear edema
Group n Increased w eigh t(x±sx , mg) Inhibi ti on rate(%)
Blank con t rol(xylene(0.01 mL , injection)+vehicle(10 mL/kg , p.o.)) 5 10.00±0.32 00.00
Posi tive cont rol(xylene(0.01 m L , injection)+diclofenac sodium (10 mg/kg , p.o.)) 5 6.50±0.24＊＊ 35.00
Test 1 (xylene(0.01 mL , injection)+plan t ext ract(250 m g/kg , p.o.)) 5 8.20±0.37＊＊ 18.00
Test 2 (xylene(0.01 mL , injection)+plan t ext ract(500 m g/kg , p.o.)) 5 7.25±0.26＊＊ 27.50
　　＊＊P<0.01 , vs b lan k cont rol g rou p.p.o.:per oral.
Table 3　Effects of ethanolic calyx extract of Hibiscus sabdari f fa on latent period
of diarrheal induction in castor oil-induced diarrheal mice
Group n
Mean latent period
(x±sx , h)
In crease in laten t
period (%)
Blank con t rol(cas tor oi l(0.5 m L , p.o.)+vehicle(10 mL/kg , p.o.)) 5 0.72±0.03 00.00
Posi tive cont rol(cas tor oil(0.5 m L , p.o.)+loperamide (50 mg/k g , p.o.)) 5 1.66±0.08＊＊ 56.62
Test 1 (castor oi l(0.5 mL , p.o.)+plant ext ract(250 m g/kg , p.o.)) 5 0.98±0.05＊＊ 26.53
Test 2 (castor oi l(0.5 mL , p.o.)+plant ext ract(500 m g/kg , p.o.)) 5 1.22±0.04＊＊ 40.98
　　＊＊P<0.01 , vs b lan k cont rol g rou p.p.o.:per oral.
Table 4　Ef fects of ethanolic calyx extract of Hibiscus sabdari ff a on frequency
of defecation in castor oil- induced diarrheal mice
Group n Mean num ber of s tools (x±sx) Inhibi ti on rate(%)
Blank con t rol(cas tor oi l(0.5 m L , p.o.)+vehicle(10 mL/kg , p.o.)) 5 17.80±1.19 00.00
Posi tive con t rol(castor oil(0.5 m L , p.o.)+l operamide(50 mg/kg , p.o.)) 5 5.21±0.89＊＊ 70.78
Test 1 (castor oi l(0.5 mL , p.o.)+plant ext ract(250 m g/kg , p.o.)) 5 12.20±1.08＊＊ 31.46
Test 2 (castor oi l(0.5 mL , p.o.)+plant ext ract(500 m g/kg , p.o.)) 5 9.60±1.22＊＊ 46.06
　　＊＊P<0.01 , vs b lan k cont rol g rou p.p.o.:per oral.
3　Discussion
　I n this study , the ethanol ic calyx extract of
H .sabdarif fa was tested to investigate the possible
ph armacological activit ies such as antinocicep-
tive , ant i-inf lammator y and antidiar rheal act ivi-
ties.An tinocicept ive act ivity was tested by using
ace tic acid-induced writhing test in mice.This
test is used for detecting both central and per iph-
er al analgesia
[ 18] . Intr aperitoneal injection of
ace tic acid causes pain and localized inf lammat ion
thr ough production of pr ostaglandins pr oduct ion ,
mainly pr ostacyclin (PGI2)and pr ostaglandin-E(PG-E), which h ave been repor ted to stimulate
the Aδ-fibres tha t cause a sensation of shar p well
localized pain
[ 20 , 21] .Ace tic acid has also been
repor ted to cause pain through act iva tion of
chemosensit ive nociceptors or irritation of visceral
surf ace th at causes to liberate histamine , br adykinin
and serotonin
[ 22] .Diclofenac sodium as a standar d
analgesic drug acts by inhibit ing the synthesis of
pr ostaglandin.Any agent th at lowers the number
of writhing will demonst rate analgesia by inhibi-
tion of prostaglandin synthesis , a peripheral
mechanism of pain inhibition.The ethanolic calyx
extract of H .sabdarif fa signif icant ly reduced the
number of writhing in mice induced by the injec-
tion of acet ic acid in a dose-dependent manner.
The refor e , the result of the acet ic acid-induced
wr ithing model mice suggests th at the extr act may
inhibi t the writhing via inhibition of prostaglandin
synthesis.　The extract was investigated to evaluate the
ant i-inflammator y activity by using the xylene-
induced ear edema mice.Xylene causes acute
inflammat ion through the histopathological changes
and resul ts in increased thickness of the ear t issues
in mice
[ 18] .I n this study the ethanolic calyx
extr act of H .sabdarif fa significant ly inhibi ted
ear edema forma tion in xylene-induced ear edema
model mice.This inhibit ion can be considered a
direct evidence suppor ting the ant i-inflammatory
eff icacy of the eth anolic calyx extr act of H .
sabdarif fa thr ough reducing vasodilation and so
that improving edematous condit ion.　Antidiar rheal act ivi ty of the ethanolic calyx
extr act of H .sabdari ffa was also invest igated by
using the castor oil-induced diarr hea model mice
and the extract showed potent antidiarrheal activity
that supports its t radit ional use in the t reatment
of diar rhea.Castor oil is known to cause diarr hea
due to its most act ive component ricinoleic acid.
The l iber ation of ricinoleic acid from castor oil by
lipase enzyme irr itates the intest inal mucosa to
cause inflamma tion and r elease of pr ostaglandin
and ni tric oxide that st imulate mot ility and secr e-
t ion of electrolyte and water
[ 23 , 24] .Seve ral other
mech anisms have been r eported to cause diarr hea
by castor oil including inhibi tion of intest inal
Na
+-K+-ATPase act ivi ty , act ivation of adenylate
cyclase or mucosal cAMP-mediated active secr e-
t ion , and platelet-activat ing factor[ 25] .The present
study showed tha t the ethanolic calyx extr act of
H .sabdari ffa possesses significant anti-inflammatory
proper ties.It is possible that the antidiarr heal
act ion exer ted by this ext ract may be rela ted to
·629·中西医结合学报 2011年 6月第 9卷第 6期　J ou rnal of C hinese Integrative Medicine , Ju ne 2011 , Vol.9 , No.6
the inhibit ion of prostaglandin format ion.However ,
confirmat ion through fur ther studies is needed
before such assert ion is made.　Preliminary phytochemical screening of ethanolic
calyx extract of H .sabdari ffa showed the presence of
reducing sugars , steroids , glycosides , alkaloids ,
saponins , flavonoids , and tannins .Alkaloids ,
flavonoids , saponins and tannins have been reported
to have mul tiple pharmacological ef fects such as
antinocicept ive
[ 26-28] , ant i-inflammator y[ 29 , 30] ,
antioxidant
[ 31]
and antidiarr heal
[ 24 , 32]
act ivi ties.
The refor e , antinocicept ive , anti-inflamma tory
and ant idiarrheal effects of the extract may be
due to the presence of flavonoids , tannins and
alkaloids either singly or in combinat ion.
4　Conclusion
　Accor ding to the above mentioned results , it
can be concluded that the e thanol ic calyx extr act
of H .sabdari f fa possesses significant antinocicep-
tive , ant i-inf lammator y and antidiar rheal act ivi-
ties that support this plant in tr adi tional medicine
use.This study also suggests f urther invest igat ion
to isolate most bioactive compounds responsible for
the uses of this plant as t radit ional medicine.
5　Acknowledgements
　Sincere thank to the Biotechnology and Genetic
Engineer ing D iscipline , Khulna Unive rsity , Ban-
gladesh f or giving financial suppor t and to the
author ity of ICDDR , B for providing the experi-
mental animals and also Bangladesh Nat ional Her-
bar ium for the ident ificat ion of the plant.
6　Competing interests
　The authors declare that they have no competing
interests .
REFERENCES
1　Mahadevan N , Shivali , Kamboj P.Hibiscuss sabdari ff a
Linn.— an overview.Nat Prod Rad.2009;8(1):77-
83.
2　Dahiru D , Obi OJ , Umaru H .Effect of H ibiscus
sabdari f f a calyx ex tr act on ca rbon tetr achloride induced
liv er damage.Biokemistri.2003;15(1):27-33.
3　Fakeye TO , Pal A , Baw ankule DU , Khanuja SP.
Immunomodulatory effect of ex tracts of Hibiscus sabdari f fa
L.(Family Malvaceae) in a mouse mode l.Phy tother
Res.2008;22(5):664 668.
4　Prommetta P , Phivthong-ngam L , Chaichantipyuth C ,
Niwa ttisaiwong N , Law anprasert S.Aqueous ex tract of
the calyces of Hibiscus subdari ff a Linn.:effects on
hepatic cy to chrome P450 and subacute tox icity in rats.
Thai J Pharm Sci .2006;30(1-2):8-18.
5　Bako IG , Mabrouk MA , Abubaka r A.Antio xidant
effect of ethano lic seed extr act of Hibiscus sabdari ff a
L inn(Malvaceae)alleviate the to xicity induced by chronic
administra tion of sodium nitrate on some haemato log ical
parameter s in Wista r rats.Adv J Food Sci Techno l.
2009;1(1):39-42.
6　Bako IG , Mohammad S T , Dawud FA , Mohammad IM ,
L iman AA.Hypotensive effect o f e thanolic seed ext ract
of H ibiscus sabdari ff a Linn (Malvaceae)on no rmo ten-
sive cats.Int J Pure Appl Sci.2009;3(3):22-28.
7　Fakeye TO , Pa l A , Khanuja SP.Anxioly tic and seda-
tive effects o f e xtr acts of Hibiscus sabdari ff a Linn
(family Malv aceae).Afr J Med Med Sci.2008;37(1):
49-54.
8　Reanmongko l W , Ithara t A.Antipy retic activ ity o f the
ex tracts of Hibiscus sabdari ff a caly ces L.in expe ri-
mental animals.Songklanakarin J Sci Technol.2007;
29(Suppl 1):29-38.
9　Fakeye T.Tox icity and immunomodula to ry activity o f
fractions of Hibiscus sabdari ff a Linn(Family Malvaceae)
in animal models.Afr J T radit Complement A ltern
Med.2008;5(4):394-398.
10　Adegunloye BJ , Omoniyi JO , Owo labi OA , Ajagbonna
OP , So fola OA , Coker HA.Mechanisms of the blo od
pressure lowe ring effect o f the caly x o f Hibiscus
sabdari f f a in rats.Afr J Med M ed Sci.1996;25(3):
235-238.
11　Chew onarin T , Kinouchi T , Kataoka K , Arimochi H ,
Kuw ahara T , Vinitke tkumnuen U , Ohnishi Y.Effect
of roseele(Hibiscus sabdari ff a Linn.), a Thai medicinal
plant , on the mutagenicity o f v arious know n mutagens
in Salmonella typhimurium and on formation of aberrant
crypt foci induced by co lon ca rcinogens azoxyme thane
and 2-amino-1-me thy l-6-phenylimidazo [ 4 , 5-b] py ridine
in F344 rats.Food Chem Toxicol.1999;37(6):591-
601.
12　Ali MB , Salih WM , Mohamed AH , Homeida AM.
Investig ation on the antispasmodic po tential o f Hibiscus
sabdari ff a caly ces.J Ethnopharmacol.1991;31(2):
249-257.
13　Olaley e MT.Cy to tox icity and antibacterial activ ity o f
methano lic ex trac t o f Hibiscus sabdari ff a.J M ed Plant
Res.2007;1(1):9-13.
14　Zimmermann M.Ethical guideline s fo r inve stigations o f
experimental pain in conscious animals.Pain.1983;16
(2):109-110.
15　Sofowo ra A.Medicinal plants and traditional medicine
in Africa.Ibadan:Spectrum Books Ltd.1993:151-153.
16　H arbo rne JB.Phy tochemical methods , a guide to modern
technique s to plant analysis.3rd ed.London:Chapman
and Hall.1998:60-66.
17　Ahmed F , Selim MS , Das AK , Choudhuri MS.Anti-
inflammatory and antinociceptive activities of L ipia
·630· 中西医结合学报 2011年 6月第 9卷第 6期　Jou rnal of Chines e Integrative M edicine , June 2011 , Vol.9 , No.6
nodiaf lora Linn.Pharmazie.2004;59(4):329-330.
18　Deb D , Dev S , Das AK , Khanam D , Banu H , Shahriar
M , Ashraf A , Choudhuri MSK , Bashe r SAMK.
Antinociceptiv e , anti-inflammato ry and anti-diar rheal
activities of the hydroalcoho lic ex tract o f Lasia spinosa
Linn.(Araceae) roo ts.La t Am J Pharm.2010;29
(8):1269-1276.
19　Chatter jee TK.H andbook o f labo rato ry mice and rats.
Calcutta:Jadavpur Univ ersity Pr ess.1993:133-139.
20　Reynolds JEF , Prasad AB.Martindale: the ex tra
pharmacopoeia.28th ed.London:The Pharmaceutical
Pre ss.1982:245.
21　Rang HP , Dale MM.Pharmaco logy.2nd ed.Edinburgh:
Churchill L ivingstone.1991:706-711.
22　Achary a SD , Ullal SD , Padiy ar S , Rao YD , Upadhyaya
K , Pillai D , Raj V.Analgesic effect of ext racts of
A lpinia galanga rhizome in mice.J Chin Integ r Med.
2011;9(1):100-104.Eng lish w ith abst ract in Chinese.
　　Acha rya SD , Ullal SD , Padiy ar S , Rao YD , Upadhyaya
K , Pillai D , Raj V.大高良姜根茎提取物对小鼠的镇
痛作用.中西医结合学报.2011;9(1):100-104.
23　Ezenw ali MO , Njoku OU , Okoli CO.S tudies on the
anti-diarrheal proper ties of seed ex trac t o f Monodora
tenui folia.Int J Appl Res Nat P rod.2009;2(4):20-26.
24　Mahesh GS , Patel P , Roy M PSP , Patel AN.Antidiar-
rheal activity o f methano lic ex tract of Moringa olei f era
Lam roo ts in experimental animal mode ls.Int J Pharm
Resea rch.2010;2(2):35-39.
25　Meite S , Nguessan JD , Bahi C , Yapi HF , Djaman
AJ , Guina FG.Antidiarrheal activity of the e thy l
aceta te ex tract o f Morinda morindoides in rats.T rop J
Pha rm Res.2009;8(3):201-207.
26　Chakrabo rthy GS , Ghorpade PM.Antinociceptive activity
of Abutilon indicum (Linn)swee t stem ex tracts.Arch
Pha rm Sci Res.2010;2(1):241-245.
27　Bhalke RD , Anar the SJ , Sasane KD , Sa tpute S N ,
Shinde SN , Sang le VS.Antinociceptive activity o f
Trigonella foenum-graecum leaves and seeds(Fabaceae).
Ir an J Pharmaco l The r.2009;8(2):57-59.
28　Zulfike r AHM , Rahman MM , Hossain MK , Hamid K ,
Mazumder M EH , Rana MS.I n vivo analg esic activ ity
of e thanolic ex tracts of two medicinal plants — Scoparia
dulcis L.and Ficus racemosa Linn.Biol Med.2010;2
(2):42-48.
29　Krishnaraju AV , Rao CBM , Sundara raju D , Sengupta
K , T rimurtulu G.Anti-inflammator y activity o f Vite x
leucoxy lon L.bark ex tracts ag ainst F reunds complete
adjuvant induced a rthritis in Sprague Daw ley rat.Am J
Infect Dis.2009;5(2):68-73.
30　Manga HM , Brkic D , Marie DE , Quetin-Leclercq J.In
v ivo anti-inflammatory activ ity o f Alchornea cordi f olia
(Schumach.&Thonn.)Mǜ ll.Arg.(Euphorbiaceae).
J Ethnopharmaco l.2004;92(2-3):209-214.
31　P reethi KC , Kut tan G , Kuttan R.Anti-inflamma to ry
activity o f f low er ex tract of Calendula off icina lis Linn.
and its possible mechanism o f action.Indian J Exp Bio l.
2009;47(2):113-120.
32　Sanni S , Thilza IB , Talle M , Mohammed SA , Sanni
FS , Okpo li LA , Jajere MS , Disa SH.The effect o f
Acacia nilotica pob ethy l acetate fraction on induced
diar rhea in albino rats.New York Sci J.2010;3(8):
16-20.
玫瑰茄花萼乙醇提取物的抗痛觉敏感 、抗炎及止泻作用的实验研究
Md.Khadem Ali1 , Ayesha Ashraf1 , Nripendra Nath Biswas2 , Utpal Kumar Karmakar2 , Shamima Afroz1
1.Bio technolog y and Genetic Engineering Discipline , L ife Science School , Khulna Univer sity , Khulna-9208 , Bang ladesh
2.Pharmacy Discipline , Life Science Schoo l , Khulna Univer sity , Khulna-9208 , Bang ladesh
目的:研究玫瑰茄(Hibiscus sabdari f f a)花萼的乙醇提取物的抗痛觉敏感 、抗炎及止泻作用 。
方法:使用 95%乙醇提取玫瑰茄花萼干品用于测定其功效。用小鼠扭体实验检测其抗痛觉敏感作用 ,二甲
苯致耳水肿模型小鼠检测其抗炎症作用 ,蓖麻油致腹泻模型小鼠检测其止泻作用。
结果:在乙酸致小鼠扭体实验中 ,玫瑰茄花萼的乙醇提取物对小鼠扭体的抑制与空白对照组相比差异有统计
学意义(P<0 .01),对二甲苯致耳水肿模型小鼠的耳水肿的抑制与空白对照组相比差异有统计学意义(P<
0.01),且显著减少了蓖麻油致腹泻小鼠的排便次数并增加了排便间隔时间(P<0.01)。
结论:本研究的结果证实了玫瑰茄花萼的乙醇提取物具有显著的抗痛觉敏感 、抗炎及止泻作用 ,验证了其在
传统医学中的应用。
关键词:木槿属;植物提取物;镇痛药;抗炎剂;止泻药;小鼠
·631·中西医结合学报 2011年 6月第 9卷第 6期　J ou rnal of C hinese Integrative Medicine , Ju ne 2011 , Vol.9 , No.6