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DOI:10.3736/ jcim20110216
http:// ww w.jcimjournal.com
Manaheji H , Jafari S , Zaringhalam J , Rezazadeh S ,
Taghizadfarid R .Ana lg esic effec ts o f me thano lic
ext racts o f the leaf o r r oo t o f Moringa olei f era on
complete F reunds adjuvant-induced a rthritis in ra ts.J
Chin Integr Med .2011;9(2):216-222.
Manaheji H , Jafari 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.
Received July 14 , 2010;accepted November 5 , 2010;
published online February 15 , 2011.
Full-tex t LinkOut at PubMed. Journal title in
PubMed:Zhong X i Y i J ie He X ue Bao.
Correspondence:H oma M anaheji , PhD , Associa te
Professor;Tel:+98-21-2387522;E-mail:hshardimanaheji
@yahoo .com
Zaringhalam J , Akba ri A , Tekieh E , Manaheji H , Rezazadeh S.
Achillea santolina reduces serum interlukin-6 level and hyperalg esia
during complete F reund s adjuvant-induced inflammation in male
Wista r ra ts.J Chin Integr Med .2010;8(12):1180-1189.
Zaringhalam J , Akba ri A , Tekieh E , M anaheji 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.
Full text available at http:// w w w .jcimjournal.com/ ar ticles/
publishA r ticles/ pdf/ 2010121497528.pdf
刘健 , 余学芳 , 纵瑞凯.新风胶囊对佐剂性关节炎大鼠行为及心
肌超微结构的影响.中西医结合学报.2008;6(6):606-610.
Liu J , Yu XF , Zong RK .Effects of Xinfeng Capsule on behavio rs
and myocardial ultra struc ture in adjuvant arthritis r ats.J Chin
Integr Med .2008;6(6):606-610.
Full text available at http:// w w w .jcimjournal.com/ ar ticles/
publishA r ticles/ pdf/ 200861932842.pdf
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Original Experimental Research 　实验论著
Analgesic effects of methanolic extracts of the leaf or
root of Moringa oleifera on complete Freund s
adjuvant-induced arthritis in rats
Homa Manaheji1 , Soheila Jafari1 , Jalal Zaringhalam1 , Shamsali Rezazadeh2 , Reza Taghizadfarid2
1.Physio lo gy Depar tment , Neuroscience Research Center , Shahid Beheshti Unive rsity of Medical Science ,
Tehran , Ir an
2.Institute of Medicinal Plants , Academic Center for Education , Culture and Research(ACECR), Tehran , Iran
Objective:Moringa oleifera (family Moringaceae)has been widely used in African folk medicine ,
and researchers have recent ly revealed its anti-inflammatory effects in human.This study aimed
to evaluate the analgesic properties of methanolic extracts of M .oleifera in complete Freunds
adjuvant (CFA)-induced arthrit is in rats.
Methods:Adult male Wistar rats , weighing 200 to 220 g , were used in this study.Adjuvant
arthritis was induced on day 0 by a single subcutaneous injection of CFA.The prepared
extracts from both the root and leaf(200 , 300 and 400 mg/kg)of M.oleifera were administered
intraperitonealy to rats in the treatment groups 0 , 3 and 6 d after CFA injection and indomethacin(5 mg/kg)was used as a positive control drug.Thermal hyperalgesia and mechanical allodynia
were evaluated for the analgesic effect 0 , 3 and 6 d after CFA injection.Combined methanolic
root and leaf extracts of M.oleifera (200 mg/kg)were also tested for the analgesic effect.
Results:The potency of the root or leaf extracts of M.oleifera (300 and 400 mg/kg)was similar to
that of indomethacin, resulted in significant reductions in both thermal hyperalgesia and mechanical
allodynia in rats with CFA-induced arthritis compared with the control group after 3 and 6 d ,
respectively (P <0 .01 or P <0 .05).Combined root and leaf extracts (200 mg/kg)of M .
oleifera resulted in a significant reduction in thermal hyperalgesia compared with the control
group after 3 and 6 d , respectively (P <0.01).Prophylactic injections of combined root and
leaf extracts of M.oleifera (200 mg/kg)resulted in a significant reduction in thermal hyperalgesia
compared with the control group , the root extracts group , and the leaf extracts group after 3
and 6 d , respectively (P <0.01).
Conclusion:The methanolic extracts of the root or leaf of M.oleifera are effective in the reduc-
t ion of pain induced by CFA in rats.A comparison of single and combination therapies of root
and leaf extracts also showed a synergistic effect on pain reduction.
Keywords:Moringa oleifera ;plant extracts;analgesics;arthritis , experimental;hyperalgesia;
Freunds adjuvant;rats
·216· 中西医结合学报 2011年 2月第 9卷第 2期　Journal of Chinese In tegrat ive Medicine , February 2011 , Vol.9 , No.2
　Moringa oleifera (Mor ingaceae family) is a
bush of the African savannah that is used in folk
medicine for the treatment of rheumat ic and ar ticular
pain.The whole plant is believed to possess
medicinal proper ties.I t is used to tr eat asci tes ,
rheumat ism , venomous bites and pneumonia[ 1] .
M .olei fera is a small- to medium-sized t ree that is
found in most western areas of Bengal.Various
parts of this plant have been used in t ribal medi-
cine.M .olei fera is commonly used in the trea t-
ment of rheumat ic swellings in t raditional medi-
cine
[ 2] .Flowers and young leaves are also consumed
for nour ishment.The methanolic extr acts of the
M .olei fera root can act as a central nervous system
depressant
[ 3] , and the aqueous root ext racts have
been shown to possess some ant i-infer til ity prop-
er ties in r ats
[ 4 , 5] .A paste of the leaves is also used
to t reat dermal wounds .Mor eover , the leaves
contain a large amount of pr oteins and are a r ich
sour ce of essential amino acids , such as methio-
nine , cyste ine , t ryptophan and lysine.The leaves
are also considered a potent ial sour ce of natural
antioxidants.Both methanolic and ethanol extractions
from the le aves have been shown to be the best
sour ce of antioxidant compounds
[ 6] .M .oleifera
not only pr oduces analgesia in mice but also
potentia tes the analgesic effects of morphine and
pe thidine
[ 2] .The macerated root of M .oleifera is
used by her bal ists in West Afr ica to tre at inflam-
mator y condit ions.The methanolic ext ract of the
root possesses ant i-inflammatory proper ties ,
which is effect ive in the t rea tmen t of both acute
and chronic inflammation
[ 3] .The composit ion
and ant i-inflammatory effects of the M .oleifera
root have been studied , but the analgesic pr oper-
ties have not yet been elucidated
[ 6] .I n recent
decades , the effects of extracts from the leaves , seeds
and roots of M .olei fera on dermal wounds[ 7] and in
pain t reatment
[ 8]
have been extensively studied .
Rheumatoid arthritis (RA), which is characterized
by chronic inflammatory changes , cart ilage destruc-
t ion , joint def ormity and disability[ 9] , is one of
the most common rheumat ic condit ions .The pain
of RA is more unbear able than any other form of
arthrit is
[ 10] .This study investigated the possible
analgesic pr opert ies of M .olei fera root or leaf
extr acts as well as a combinat ion of the two in male
rats wi th complete Freunds adjuvan t (CFA)-
induced arthr it is.
1　Materials and methods
1.1　Plant materials
1.1.1 　Extract preparation 　African generat ion
of M .oleifera was collected between June and
July of 2009.The specimens were ident ified in the
Centr al Her bar ium of I ran (Central Herbar ium of
Medicinal Plants , Academic Center for Educa-
tion , Culture and Research , Tehran , Ir an).The
vouche r specimen was deposited the re with No .
66.The plant mater ials were cleaned and shade
dried prior to extraction.Analytical grade solvents
were used for extr action .Quercet in was used as
standard flavonoid (Sigma-Aldrich , St .Louis ,
Missour i , USA);solvents for chroma tographic
analyses were obtained from Panreac Química S.
A.U., Spain;minor chemicals , such as acetic acid
and anhydrous sodium sulfate , were obta ined fr om
Merck &Co., Inc.The aer ial port ion and roots(200 g)of the plant were subjected to macer ation
with methanol(99.9%)for 72 h , and the methanolic
extr acts wer e then filtered .The extracts we re
then evapor ated in vacuum a t the temper ature of
about 40 ℃.The yields for the methanolic extracts
were determined , and the dried extracts were kept
at 4 ℃ until use.By using Tween 80 as solubilizing
agent , the dried extr acts were dissolved in ster ile
water containing 0.9 % (w/v)sodium chloride
and passed through a weighed paper f ilter .
1.1.2 　Phytochemical analyses 　Standard phyto-
chemical screening tests
[ 10]
were used to screen the
extr acts .The plants wer e qualitat ively analyzed
to de tect saponins , alkaloids and ter penoides.For
quant izat ion of flavonoids , the following high-
performance liquid chr omatogr aphy (HPLC)
inst rument was used:KNAUER Chr omGate V3.
1.7 softwar e , pump K-1001 , solvent or ganizer K-
1500 (KNAUER , Germany);ODS C18 column(30 cm)and L1 (spher ical , d =5 μm).Chroma-
togr aphy was perf ormed at 30 ℃with a flow ra te
of 1.5 mL/min , and the mobile phase was methanol(77%) and water-phosphor ic acid (1 ∶ 200)(23%).The ultraviolet detector was set at 225 nm.
Extr acts were prepar ed with a minor change to
the previously described method
[ 10] .About 1.0 g
of the plant , f inely pulverized and accur ately
weighed , was tr ansferred to a 250 mL flask fit ted
with a reflux condenser , and 78 mL of extr action
solvent (alcohol , water , and hydrochloric acid(50 ∶20 ∶8 , v/v/v))was then added , and the
solution was refluxed in a hot water bath for about
135 min.The solut ion was cooled to room tem-
perature and decanted into a 100 mL volumetric
flask , and 20 mL of methanol was then added to
the 250 mL flask , and the solution was sonicated
for 30 min , fil tered and collected in a 100 mL
volumetric flask .The residue on the fil ter was
washed with me thanol , and the flow-through was
collected in the same 100 mL volumetric flask ,
diluted and mixed .A total of 20 μL of this solu-
t ion was injected for HPLC analysis.Prior to
analysis , a cal ibr ation cur ve was prepared with
0.01 , 0.02 , 0.05 , 0.10 and 0.20 mg/mL concen-
t rat ions of quercet in as the standard .Flavonoid
concentrat ions were calculated according to an
establ ished method , and the data wer e presented
as que rce tin per centage (w/w)of plant material.
1.2　Laboratory animals　Adult male Wistar rats ,
weighing 200 to 220 g , wer e used in this study.
The rats were housed in individual cages with f ree
access to food and water.They were kept in a
·217·中西医结合学报 2011年 2月第 9卷第 2期　J ou rnal of C hinese Integrative Medicine , Feb ruary 2011 , Vol.9 , No.2
tempe rature-contr olled room at (22.0 ±0.5) ℃
with a 12/12 h light/dark cycle (l ights on from
06:00 to 18:00).The humidity of the r oom was
kept between 60 % and 62%.The study protocol
was approved by the local e thics commit tee for
the use of animals in research , and we f ollowed
the guidelines of e thical standards for the invest i-
gat ion of experimental pain in animals
[ 8] .
1.3　Experimental methods
1.3.1 　Induction of adjuvant arthritis 　Adjuvant
ar thritis was induced on day 0 by a single subcutaneous
injection into the righ t hindpaw of 100 μL of
heat-killed Mycobacterium tuberculosis suspended
in sterile mine ral oil (10 mg/mL CFA , Sigma
Chemical Co., St Louis , Missouri , USA).The
right hindpaws of sham rats were injected with
ster ile mineral oil only (100 μL)[ 9] .Hindpaw
unilateral edema (acute phase)was established
within the first hour after CFA inject ion , and it
was quant ified during the first week
[ 11] .
1.3.2 　Assessment of thermal hyperalgesia　Thermal
hyperalgesia was evaluated by using the pr eviously
described r adiant heat me thod.Rats were placed
in plexiglas boxes for 10 to 15 min prior to testing
to h abitua te to the test environment.Paw with-
drawal latency(PWL)in response to r adiant heat
was measured with a plantar test appara tus (Ugo
Basile , I taly).The heat sour ce was positioned
under the plantar surface of the af fected hindpaw
and act ivated.A digital t imer connected to the
heat source automat ically recorded the PWL to a
tenth of one second .If the r at did not withdr aw
its paw from the stimulus after 20 s , the test was
terminated , and the r at was assigned with this
cut-of f value.Each r at underwent three t rials per
hindpaw at intervals of 5 to 10 min .The mean
latency of the withdr awal responses for each paw
was calculated.The value for the right (CFA-
injected)paw was then subtracted from the value
for the lef t paw , and the resul t was considered as
a measure of hyperalgesia in the injured paw
[ 12] .
1.3.3 　Mechanical allodynia 　To evalua te the
contr ibut ion of low-threshold f ibers to nocicept ive
behavior , we studied the effects of st imulat ion
with weak von Frey f ilaments with bending for ces
ranging from less than 2 to 60 g (Stol ting Inc.,
Wood Dale , Ill inois , USA).This st imulus is
consider ed innocuous because it normally elicits
activity in low-threshold mechanoreceptors.Rats
wer e placed on a mesh (0 .8 cm ×0 .8 cm)floor ,
covered with an inver ted tr ansparent plastic box(20 cm×20 cm ×18 cm)and allowed to adapt for
approximately 15 min or until exploratory behavior
ceased .The centr al region of the plantar surf ace
of the hindpaw was stimula ted with a series of von
Frey filaments in ascending or der of for ce.The
paws we re st imulated three t imes consecutively by
pushing up on the hindpaw unt il the rat ei ther
withdrew its paw or the fiber bowed .Lift ing of
the paw due to normal locomotor behavior was
ignored .The withdr awal threshold represented
the smallest filament size that evoked at least two
withdr awal responses during three consecutive
applicat ions of the same f ilament .Each f ilament
was applied f or approxima tely 1 s , and the inter-
st imulus interval was appr oximately 5 s[ 13] .
1.3.4　Experimental procedure　To determine the
effects of M .oleifera r oot or leaf ext racts on
CFA-induced pain , a ser ies of experiments we re
performed on rats , which were divided into various
exper imental groups (n =6 per group).Treat-
ments with M .olei fera r oot or leaf extr acts we re
administered 0 , 3 and 6 d afte r CFA injection.
Thermal hyperalgesia and mech anical allodynia
were measur ed 0 , 3 and 6 d af ter CFA injection.
In the next part of the study , r ats received com-
bined meth anolic root and le af ext racts 0 , 3 and
6 d after CFA inject ion .Thermal hyperalgesia
and mechanical allodynia were assessed 30 min
befor e CFA inject ion and 3 and 6 d af ter adminis-
t rat ion of M .olei fera extracts.Carboxy methyl
cellulose(CMC)was used as a vehicle for dilution of
the extracts and the drug.Indomethacin (5 mg/kg),
a standard analgesic agent , was used as a positive
con trol.
1.4 　Statistical analysis　Data were pr esented as
x ±s x .One way analysis of variance (ANOVA)
followed by post hoc Tukeys multiple comparison
test (Statist ica 6.0), and unpaired student t-test
were used to de termine signif icant dif ferences in
thermal and mechanical pain thresholds within
and between groups wher e appropriate.Stat ist ical
signif icance was accepted at P <0.05 .
2　Results
2.1　The yields of extracts　For the ae rial par ts of
the plant , the yields for methanolic and def at ted
extr acts were calculated to be 16.2%(w/w)and
10 .3 %(w/w), respect ively.For the r oots , the
methanolic and defatted yields were 11.7%(w/w)
and 6.4%(w/w), respectively.
2.2 　Phytochemical analyses results　Both the aerial
par ts and the roots con tained saponins and we re
rich in terpenoids.However , alkaloids were absent.
The cal ibr ation curve showed good l inearity with
a regression value of 0.995 1.The total flavonoids
con tent as a per centage of quer ce tin content was
0.65%(w/w).
2.3　Effects of M.olei f era leaf extracts　Treatment
with M .oleifera leaf extracts (300 and 400 mg/kg)
resulted in signif icant reduct ions in both thermal
hyper algesia and mechanical allodynia af ter 3 and
6 d in ra ts wi th CFA-induced arthrit is compared
with the control group (P <0.01 or P <0.05).
Effect ive doses of me thanol ic leaf extr acts (300
and 400 mg/kg) significantly reduced hyperalgesia
and mechanical allodynia on day 6 compared with
day 3 in CFA-injected r ats(P <0.05).There were
no significant differences in thermal hyperalgesia and
mechanical allodynia in r ats treated with 200 mg/kg
·218· 中西医结合学报 2011年 2月第 9卷第 2期　Journal of Chinese In tegrat ive Medicine , February 2011 , Vol.9 , No.2
of M .oleifera le af extr acts compared with the
control group.Thermal hyperalgesia and mechanical
allodynia were significant ly reduced by tre atment
with 5 mg/kg of indomethacin (5 mg/kg)com-
par ed with the control group (P <0.05 or P <
0.01).There were no significant differences between
rats treated with effective doses of the leaf extr acts(300 and 400 mg/kg)and rats treated with indom-
ethacin (5 mg/kg)(Table 1).
2.4　Effects of M.olei f era root extracts　Compared
with the contr ol group , ra ts with CFA-induced
arthrit is that were treated with either 300 mg/kg or
400 mg/kg of M .olei fera r oot ext racts showed a
significant reduct ion in thermal hyperalgesia af ter
3 and 6 d treatment (P <0.01).Thermal hyperalgesia
signif icantly reduced a t day 6 compared with day
3 in 300 and 400 mg/kg M .olei fera root ext racts-
t reated groups.However , no significant differ-
ences in mechanical allodynia were obser ved , and
200 mg/kg treatment h ad no effect on either
variable.I ndomethacin (5 mg/kg)t reatment also
signif icantly r educed thermal hyperalgesia (P <
0.01) at day 3 and day 6 af ter CFA injection.
There were no significant differences in thermal
hyper algesia and mechanical pain thresholds
between rats t reated with effective doses of root
extr acts (300 and 400 mg/kg) and rats tre ated
with indometh acin (5 mg/kg)(Table 2).
Table 1　Effects of Moringa oleif era leaf extracts on CFA-induced thermal hyperalgesia and mechanical allodynia(x ±sx)
G roup n
Mean value of w ithdrawal latencies
betw een right and left hindpaw s (second) Mean valu e of threshold (g)
C on t rol
　0 d of t reament 6 0.17±0.05 60.00±1.24
　3 d af ter t reament 6 -6.38±0.25 15.14±1.32
　6 d af ter t reament 6 -5.05±0.35 12.14±2.08
M ethanolic ext ract s(200 mg/kg)
　0 d of t reament 6 0.24±0.08 60.00±1.85
　3 d af ter t reament 6 -6.43±0.35 15.29±2.05
　6 d af ter t reament 6 -4.90±0.44 15.14±1.55
M ethanolic ext ract s(300 mg/kg)
　0 d of t reament 6 0.18±0.05 60.00±1.58
　3 d af ter t reament 6 -4.33±0.31＊＊ 29.15±1.55＊＊
　6 d af ter t reament 6 -3.90±0.09■△ 20.00±1.55■△
M ethanolic ext ract s(400 mg/kg)
　0 d of t reament 6 0.25±0.1 60.00±2.11
　3 d af ter t reament 6 -4.50±0.23＊＊ 30.25±1.43＊＊
　6 d af ter t reament 6 -3.45±0.15■◆ 22.00±1.55■◆
Ind om ethacin (5 m g/kg)
　0 d of t reament 6 0.15±0.08 60.00±2.14
　3 d af ter t reament 6 -4.85±0.30＊＊ 22.94±1.13＊
　6 d af ter t reament 6 -3.10±0.35■■ 19.65±1.51■
　　＊P<0.05 , ＊＊P<0.01 , vs cont rol group 3 d af ter t reatment;■P <0.05 , ■■P<0.01 , vs cont rol group 6 d af ter treatment;△P <0.05 ,
vs Mor ing a olei fera leaf ext ract s (300 m g/kg)3 d af ter t reatm ent;◆P <0.05 , vs Mor in ga olei fera leaf ex t ract s (400 m g/kg)3 d after
t reatm ent.CFA:com plete Freunds ad juvan t.
Table 2　Effects of Moringa oleif era root extracts on CFA-induced thermal hyperalgesia and mechanical allodynia(x ±sx)
G roup n
Mean value of w ithdrawal latencies
betw een right and left hindpaw s (second) Mean valu e of threshold (g)
C on t rol
　0 d of t reament 6 0.17±0.05 60.00±2.41
　3 d af ter t reament 6 -6.25±0.42 23.35±1.42
　6 d af ter t reament 6 -5.05±0.54 20.50±1.44
M ethanolic ext ract s(200 mg/kg)
　0 d of t reament 6 0.09±0.02 60.00±2.32
　3 d af ter t reament 6 -5.80±0.21 26.00±1.55
　6 d af ter t reament 6 -4.80±0.38 23.45±1.65
M ethanolic ext ract s(300 mg/kg)
　0 d of t reament 6 0.12±0.03 60.00±2.88
　3 d af ter t reament 6 -2.34±0.25＊＊ 22.43±1.31
　6 d af ter t reament 6 -1.40±1.09■■△△ 22.45±1.39
M ethanolic ext ract s(400 mg/kg)
　0 d of t reament 6 0.20±0.05 60.00±2.11
　3 d af ter t reament 6 -2.66±0.56＊＊ 25.54±1.55
　6 d af ter t reament 6 -1.20±0.25■■◆◆ 26.55±1.11
Ind om ethacin (5 m g/kg)
　0 d of t reament 6 0.15±0.04 60.00±2.14
　3 d af ter t reament 6 -2.95±0.30＊＊ 28.65±1.30＊
　6 d af ter t reament 6 -1.54±0.51■■□□ 25.50±1.51■
　　＊P<0.05 , ＊＊P <0.01 , vs cont rol group 3 d af ter t reatment;■P<0.05 , ■■P <0.01 , vs con t rol grou p 6 d af ter t reatment;△△P <
0.01 , vs Morin ga o lei fera root ext ract s (300 m g/kg)3 d af ter t reatmen t;◆◆P<0.01 , vs Mor in ga olei fera root ext ract s (400 m g/kg)3 d
af ter t reatmen t;□□P <0.01 , vs indometh acin(5 mg/kg)3 d af ter t reatment.CFA:com plete Freunds adjuvan t.
·219·中西医结合学报 2011年 2月第 9卷第 2期　J ou rnal of C hinese Integrative Medicine , Feb ruary 2011 , Vol.9 , No.2
2.5　Effects of combined M.olei f era leaf and root
extracts 　Treatment with combined M .oleifera
leaf and root ext racts (200 mg/kg)resulted in a
dr ama tic decrease in thermal hyper algesia (P <
0.01)on day 3 and day 6 in rats with CFA-induced
arthrit is .However , ther e were no significant
effects on mechanical allodynia compared with the
control group.There were no significant differences in
thermal hyperalgesia and mechanical allodynia
between the group which administered combined
M .oleifera le af and root ext racts (200 mg/kg)
and indome thacin-tre ated group a t different days
of the study (5 mg/kg)(Table 3).
2.6 　Prophylactic effects of combined M.olei f era
leaf and root extracts　Pr ophylact ic administ rat ion
of combined M .oleifera le af and root extracts(200 mg/kg)resul ted in a dramatic decrease in
thermal hype ralgesia on day 3 and day 6 in rats
with CFA-induced ar thri tis in compar ison with
the control group , the root ext racts group , and
the le af ext racts group , respectively (P <0.01).
Combined extracts-treated group showed no significant
difference in thermal hyperalgesia between day 3
and day 6.Ther e wer e no signif icant differences
in mech anical allodynia between the different
exper imental groups .There was no significant
difference in mech anical allodynia between the
gr oup tr eated with combined M .oleifera leaf and
root ext racts (200 mg/kg)and the group tre ated
with indometh acin (5 mg/kg)(Table 4).
Table 3　Effects of combinedMoringa oleif era leaf and root extracts on CFA-induced thermal hyperalgesia and mechanical allodynia(x ±sx)
G roup n
Mean value of w ithdrawal latencies
betw een right and lef t hindpaw s(secon d) Mean valu e of threshold (g)
C on t rol
　0 d of t reament 6 0.17±0.06 60.00±1.24
　3 d af ter t reament 6 -6.38±0.25 17.00±1.45
　6 d af ter t reament 6 -5.05±0.35 21.84±2.11
M ethanolic leaf ext ract s (200 m g/kg)
　0 d of t reament 6 0.24±0.11 60.00±1.85
　3 d af ter t reament 6 -6.43±0.35 17.29±1.15
　6 d af ter t reament 6 -4.90±0.44 23.14±1.45
M ethanolic root ext ract s (200 mg/k g)
　0 d of t reament 6 0.09±0.02 60.00±2.32
　3 d af ter t reament 6 -5.80±0.21 17.05±1.55
　6 d af ter t reament 6 -4.80±0.38 23.45±1.65
C om bined methanolic ex tract s (200 m g/kg)
　0 d of t reament 6 0.41 ± 0.1 60.00±2.11
　3 d af ter t reament 6 -2.62 ± 0.55＊＊ 19.25±1.45
　6 d af ter t reament 6 -2.91 ± 0.45■■ 24.15±1.35
Ind om ethacin (5 m g/kg)
　0 d of t reament 6 0.52±0.14 60.00±2.14
　3 d af ter t reament 6 -3.11±0.30＊＊ 21.54±1.24＊
　6 d af ter t reament 6 -3.24±0.51■■ 25.15±1.51■
　　＊P<0.05 , ＊＊P <0.01 , vs cont rol group 3 d af ter t reatment;■P <0.05 , ■■P<0.01 , vs cont rol group 6 d af ter t reatment.CFA:
complete Freunds adjuvant.
Table 4　Prophylactic ef fects of combined Moringa olei f era leaf and root extracts on
CFA- induced thermal hyperalgesia and mechanical allodynia (x ±sx)
G roup n
Mean values of wi thd raw al latencies
b etw een right and lef t hindpaw s(second) Mean values of th resh old(g)
C on t rol
　0 d of t reament 6 0.17±0.08 60.00±1.25
　3 d af ter t reament 6 -6.01±0.35 21.00±1.34
　6 d af ter t reament 6 -5.20±0.35 23.44±1.41
M ethanolic leaf ext ract s (200 m g/kg)
　0 d of t reament 6 0.24±0.09 60.00±1.65
　3 d af ter t reament 6 -6.34±0.35 23.25±1.32
　6 d af ter t reament 6 -4.54±0.44 23.90±1.35
M ethanolic root ext ract s (200 mg/k g)
　0 d of t reament 6 0.29±0.10 60.00±1.65
　3 d af ter t reament 6 -5.85±0.21 22.85±1.45
　6 d af ter t reament 6 -4.65±0.38 24.47±1.65
C om bined methanolic ex tract s (200 m g/kg)
　0 d of t reament 6 0.32±0.12 60.00±2.12
　3 d af ter t reament 6 -1.55±0.31＊＊☆☆★★ 23.05±1.41
　6 d af ter t reament 6 -1.00±0.19■■ 25.95±1.55
Ind om ethacin (5 m g/kg)
　0 d of t reament 6 0.62±0.14 60.00±2.14
　3 d af ter t reament 6 -1.31±0.30＊＊ 25.56±1.34＊
　6 d af ter t reament 6 -1.54±0.51■■ 27.85±1.51■
　　＊P<0.05 , ＊＊P <0.01 , vs cont rol group 3 d af ter t reatment;■P<0.05 , ■■P <0.01 , vs con t rol grou p 6 d af ter t reatment;☆☆P <
0.01 , vs Moringa olei fera leaf ext racts (200 mg/kg)3 d after t reatment;★★P <0.01 , vs Moringa olei f era root ex tract s (200 mg/k g)3 d
af ter t reatmen t; P<0.01 , vs Moringa olei fera leaf ex t ract s (200 mg/kg)6 d af ter t reatm en t; P <0.01 , vs Mor ing a olei fera root ext racts
(200 mg/kg)6 d af ter t reatm ent.CFA:com plete Freunds ad juvan t.
·220· 中西医结合学报 2011年 2月第 9卷第 2期　Journal of Chinese In tegrat ive Medicine , February 2011 , Vol.9 , No.2
3　Discussion
　As the most important pharmacological model
of RA , rat adjuvant arthrit is mimics the ar thr itis
observed in humans.This rat model was used to test
the classic non-steroidal anti-inflammatory drugs(NSAIDs)[ 14] .CFA inject ion into the paws of the
rats pr ovokes local in flamma tion and pa in .
Tr eatment wi th methanolic extr acts fr om the
leaves of M .olei fera r esulted in a significant
decrease in the rmal hyper algesia and mechanical
allodynia on day 3 and day 6 after CFA inject ion .
Treatment with methanolic extracts from the roots of
M .olei fera resul ted in a significant decrease in
thermal hyperalgesia but had no signif icant e ffect
on mechanical allodynia at 300 and 400 mg/kg
either on day 3 and day 6 af ter CFA inject ion .
These effects were comparable with the effects of
5 mg/kg of indomethacin.Gupta et al [ 2] studied the
analgesic ef fects of the M .olei fera root at doses
ranging from 350 to 700 mg/kg using the hot plate
method.They have shown that the root of M .
olei fera not only produced analgesia in mice but also
potentiated the analgesic actions of morphine and
pe thidine.Ezeamuzie et al [ 3] showed that the M .
oleifera root had ant i-inflammatory ef fects on
paw edema after 6 d in the rat air pouch inflammatory
model.The phytochemical screening of the leave
and root extr acts of M .oleifera indicated the
pr esence of various classes of chemicals , such as
ter penoids , saponin glycosides and flavonoids.
Flavonoids , such as quercet ins , are known to be
effective in reducing inflammatory symptoms.
These flavonoids possess potent inhibitory effects
on various enzymes , such as prote in kinase C ,
phospholipase A2 and phosphodiester ase
[ 15] .The
aur antiamide ace ta te isolated fr om the r oot of M .
oleifera has been shown to significant ly inhibit
inflammatory arthritis and analgesic activities during
the hot plate test
[ 16] .The alcohol ic extr acts of M .
oleifera seeds have been shown to have analgesic
act ivit ies compa rable to those of aspir in a t
25 mg/kg[ 1 7] .C ce res et al [ 18] ident ified ant i-
inflammator y pr opert ies of 1 000 mg/kg of the
root ext racts of M .olei fera in r odents with
carrageenan-induced edema.The leaves of M .
olei fera , especially the meth anolic and ethanolic
ext racts , are good sour ces of natur al ant ioxidant
compounds
[ 1 9] .With respect to the different
compounds in the le aves , roots and other parts of
M .oleifera that contain different substances
capable of reducing inflammat ion and pain , we
examined a combinat ion of the extr acts from the
leaves and roots (200 mg/kg).Treatment with
the combined extracts resul ted in a significant
reduction in thermal hyperalgesia but had no
significant e ffect on mechanical allodynia in
compar ison with 5 mg/kg of indomethacin .In
conclusion , the methanolic ext racts of M .oleifera
root and leaf seem to be eff ective in r educing pain
in rats with CFA-induced arthrit is .A compar ison
of the effects of single and combinat ion ther apy
with r oot and leaf extracts showed the synergist ic
act ivi ty of both in the reduct ion of pain .This
may further suggest that , single or combinat ion of
root and leaf ext racts of M .olei fera along with
other an algesic drugs can be useful in alleviation
of inflammatory pain .
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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
Homa Manaheji1 , Soheila Jafari1 , Jalal Zaringhalam1 , Shamsali Rezazadeh2 , Reza Taghizadfarid2
1.Physio lo gy Depa rtment , Neuroscience Research Cente r , Shahid Behe shti Univ ersity of Medical Science , Tehran , Iran
2.Institute o f Medicinal Plants , Academic Center for Educa tion , Culture and Resea rch(ACECR), Tehran , Iran
目的:辣木(Moringa olei f era)是非洲民间医学中一种常用的药用植物 ,近期已有临床研究揭示了其抗炎功
效。本实验研究辣木叶及根的甲醇提取物对弗氏佐剂致关节炎模型大鼠的止痛作用 。
方法:使用健康雄性成年Wistar大鼠(体质量 200 ～ 220 g),皮下注射弗氏佐剂建立实验性关节炎模型 。造
模当天及造模后第 3 天和第 6 天 ,治疗组大鼠分别腹腔内注射辣木根或叶的甲醇提取物(200 、300 或
400 mg/kg),使用消炎痛(5 mg/kg)作为阳性对照药 。每次用药后评估各组大鼠的温度痛觉过敏程度及机
械性触诱发痛程度。实验用同样方法检测了辣木根和叶的甲醇提取物混合物(200 mg/kg)的止痛作用 。
结果:辣木根或叶的甲醇提取物(300或 400 mg/kg)的止痛作用与消炎痛(5 mg/kg)类似 ,与对照组比较 ,
造模后第 3天或第 6天用药都能够显著降低弗氏佐剂致关节炎大鼠的温度痛觉过敏程度及机械性触诱发痛
程度(P <0.01或 P <0.05)。与对照组 、辣木根提取物组及辣木叶提取物组比较 ,造模后第 3天或第 6 天用
药 ,辣木根和叶的甲醇提取物混合物(200 mg/kg)能够显著降低弗氏佐剂致关节炎大鼠的温度痛觉过敏程
度(P <0 .01)。
结论:辣木根或叶的甲醇提取物能够有效降低弗氏佐剂致关节炎大鼠的温度痛觉过敏程度。辣木根和叶的
甲醇提取物混合物与根或叶单独的提取物的作用比较显示出辣木根与叶在止痛效果上的协同作用 。
关键词:油椒木;植物提取物;镇痛药;关节炎 , 实验性;痛觉过敏;Freund佐剂;大鼠
·222· 中西医结合学报 2011年 2月第 9卷第 2期　Journal of Chinese In tegrat ive Medicine , February 2011 , Vol.9 , No.2