全 文 ： 2009 年 7 月 第 7 卷 第 4 期 Chin J Nat Med July 2009 Vol. 7 No. 4 307








Anti-inflammatory and Analgesic Effects of Two
Extracts Isolated from Illicium

ZHAO Chuan1, HE Ling1*, ZHANG Lu-Yong2, SHAN Mu1
1Department of Pharmacology, China Pharmaceutical University, Nanjing 210009;
2 National Drug Screening Laboratory, China Pharmaceutical University, Nanjing, 210009, China
[ABSTRACT] AIM: Since Illicium lanceolatum A. C. Smith is one of the endangered wildlife in China, searching for
its substitutes is quite necessary. The aim of our study was to compare the anti-inflammatory and analgesic effects of
two injections of extracts isolated from the root bark of Illicium angustisepalum A. C. Smith and Illicium Jiadifengpi
B.N. Chang, respectively. METHODS: Rat paw oedema and mice ear edema test, as well as acetic acid-induced vascu-
lar permeability, were used to evaluate the anti-inflammatory activity between the two injections. Hot-plate test and ace-
tic acid-induced writhing test were used to determine analgesic effects between the two injections. RESULTS: Both in-
jections displayed marked anti-inflammatory and analgesic effects. No significant difference between the two injections
was seen according to the statistical data. CONCLUSION: The injection of extract isolated from the root bark of Il-
licium Jiadifengpi B.N. Chang has better anti-inflammatory and analgesic effects to Illicium angustisepalum A. C. Smith.
The results indicate that Illicium Jiadifengpi B.N. Chang might possibly be used as a substitute of Illicium angustise-
palum A. C. Smith.
[KEY WORDS] Magnoliaceae; Illicium lanceolatum; Illicium Jiadifengpi; Anti-inflammatory activity; Analgesic effects
[CLC Number] R965 [Document code] A [Article ID] 1672-3651(2009)04-0307-05
doi: 10.3724/SP. J. 1009.2009.00307
1 Introduction
The genus Illicium comprises 15 species belonging to
the medicinal plants distributed mainly in China. Some of
them were described in the Chinese Medicine book, ‘Com-
pendium of Materia Medica’ as early as 1593. The root bark,
fruits and leaves of Illicium medicinal plants were used for
treating rheumatic arthritis, fractures, bleeding wounds,
abdominal distension and emesis in China for a long time[1].
Japanese scientists firstly separated shikimic acid from the
fruits of Illicium verum Hook.F. in 1885. Shikimic acid had
been shown to play an unique role in curing the patients
infected with avian influenza (AI)[2]. Other species of the
genus Illicium had also been used in clinical treatments.
Illicium lanceolatum A. C. Smith is a perennial herb
indigenous to the damp forests, which grow on the hill or
mountains at an altitude ranging from 100 to 1 600 meters.
It was an evergreen plant of about 3 to 8 meters in height,
with opposite, glabrous leaves. The purified extract isolated

[Received on] 01-Dec-2008
[Foundation Item] This project was supported by Specific
Fund for Public Interest Research of Traditional Chinese Medicine,
Ministry of Finance (No. 200707008) and the Drug Efficacy Study
and Evaluation Service Center of Jiangsu Province (BM2005103)
[*Corresponding author] HE Ling, Prof. E-mail: hel-
ing92@hotmail.com
from the root bark of Illicium lanceolatum A. C. Smith
(hereinafter referred to as Honghuixiang injection, HI) has
been used for treating edema, rheumatic and rheumatoid
arthritis clinically since 2001. Its main chemical constitu-
ents are flavonoids. Since Illicium lanceolatum A. C. Smith
is one of the endangered wildlife in China, its use has been
limited and controlled by local government, therefore,
searching for its substitutes is quite necessary.
Illicium Jiadifengpi B.N. Chang, which distributed in
45 counties in China, has been often confused with Illicium
lanceolatum A. C. Smith due to great similarities in their
appearances. The decoction of the root bark of Illicium
Jiadifengpi B.N. Chang has also been used for the treatment
of rheumatic arthralgia, aching of the loins and knees and
traumatic injuries in folk[3]. In the present study,
anti-inflammatory and analgesic effects of two extracts
isolated from Illicium angustisepalum A. C. Smith and Il-
licium Jiadifengpi B.N. Chang, respectively, were com-
pared in preclinical animal models, to investigate whether
the latter could be used as the possible substitute of the
former.
2 Materials and Methods
2.1 Medicine and agents
Carrageenan and Evans blue were purchased from
Sigma Co.. Diclofenac sodium injection was used as posi-
ZHAO Chuan, et al. /Chinese Journal of Natural Medicines 2009, 7(4): 307−311
308 Chin J Nat Med July 2009 Vol. 7 No.4 2009 年 7 月 第 7 卷 第 4 期

tive drug and obtained from Guangzhou Mingxin Pharmacy
Co., Ltd. (Guangzhou, China). Diclofenac sodium injection
was diluted using physiological saline to 0.31mg·mL−1.
All other chemicals were of reagent grade and commer-
cially available.
2.2 Plant material and extracts preparation
The purified extract isolated from the root bark of Il-
licium lanceolatum A. C. Smith (Honghuixiang injection,
HI) and the extract isolated from Illicium Jiadifengpi B.N.
Chang (IJ injection) were supplied by Zhejiang Taikang
Pharmaceutical Co., Ltd. The root barks of Illicium lanceo-
latum A. C. Smith and Illicium Jiadifengpi B.N. Chang
were collected in March 2007 from Zhejiang province of
China. Two extracts isolated from the root barks of Illicium
lanceolatum A. C. Smith and Illicium Jiadifengpi B.N.
Chang, respectively, were prepared according to the Phar-
macopoeia of China (Chinese Pharmacopoeia Committee,
Publishing House of People’s Health, 2005, page 64).
Briefly, the root barks were washed and immersed in 75%
ethanol solution at 25°C for 7-10 days, and then filtered.
The supernatant was mixed with physiologic saline, the
aqueous solution of sodium hydroxide and tween-80, then
sterilized and encapsulated into ampoules (2mL per am-
poule). Analysis was performed using high performance
liquid chromatography (HPLC) according to standard
specifications of traditional Chinese medicines. 0.1%
H3PO4, and acetonitrile were used as eluents. The concen-
tration of quercitrin, a main component in the extracts, was
required to be higher than 0.3 mg·mL−1.
2.3 Animals
Male Wistar rats weighing 220-250 g and ICR mice
of both sexes weighing (22±2) g were used. Animals were
purchased from the Experimental Animal Center of Nan-
tong University, and were kept in polyethylene boxes under
controlled temperature (22±2)°C, humidity (55±5) % and
circadian cycle, with free access to standard food and water.
The study was authorized by the Ethics Committee of China
Pharmaceutical University, in accordance with the Guide
for the Care and Use of Laboratory Animals published by
the United States National Institutes of Health.
Animals were randomly divided into groups in each
test: (1) control group: normal saline treatment; (2) positive
drug group: diclofenac sodium solution treatment; (3) ex-
tract isolated from Illicium lanceolatum A. C. Smith group:
subcutaneously administered three doses (15, 7.5, 3.75
mL·kg−1) of HI, once a day for three days; (4) extract iso-
lated from Illicium Jiadifengpi B.N. Chang group: subcuta-
neously administered three doses (15, 7.5, 3.75 mL·kg−1) of
IJ, once a day for three days. Each group included ten ani-
mals. After the final administration of extracts on the third
day, each test was performed.
2.4 Methods
2.4.1 Paw edema test After the final administration of
extracts, 0.1ml of 1% carrageenan dissolved in normal sa-
line was subcutaneously injected into the right hind paw
plantar surface of rats[4]. And after wating for 0.5, 1, 2, 4
and 6 h, respectively, the paw volume was measured using a
plethysmometer. Paw edema was evaluated by the delta
volume (a-b), where a and b was the volume of the right
hind paw after and before the carrageenan treatment, re-
spectively[5].
2.4.2 Ear edema test After the final administration of
extracts, each mouse was received 20µL of xylene to the
inner and outer surface of the right ear. The left ear re-
mained untreated. Three hours later, mice were sacrificed
by cervical dislocation and their ears were cut-off. A stain-
less steel punch with diameter of 9 mm was used to take the
central sections of the treated and untreated ears[6,7]. The
sections of the ears were weighed; weight difference be-
tween the right and left ear was used to evaluate ear edema.
The inhibition of edema in percentage was calculated using
the following formula: (edema control – edema drugs) / edema
control ×100%
2.4.3 Acetic acid-induced vascular permeability One
hour after the final administration, each mouse was intra-
venously injected with 0.1ml of 1 % evans blue dye solu-
tion and intraperitoneally injected with 0.7 % acetic acid
(0.1 mL/10 g). 30 minutes later, mice were sacrificed and
then the peritoneal exudates were collected by washing the
peritoneum with 5 mL normal saline after the viscera were
exposed. The wash solution was centrifuged at 3 000×g for
15 min. The vascular permeability was expressed as the
absorbance of the supernatant at 590 nm[8].
2.4.4 Hot-plate test Animals were habituated twice to
the hot-plate in advance. For testing, female mice were
exposed to the hot plate apparatus maintained at (55 ± 0.5)
°C. The time that elapsed until the occurrence of either a
hind paw licking or a jump off the surface was recorded as
the latency of pain response. The cut-off time was set at 60
s. Mice with latencies of <5 s or >30 s were eliminated
from the study. After the determination of the pain response
latency, measurements were performed at 30, 60 and 90min
after the final administration, respectively[9].
2.4.5 Abdominal constriction test One hour after the
final administration of the drugs, each mouse was intrap-
eritoneally injected with 0.7% acetic acid (10 mL·kg−1).
The number of writhing that occurred within the first 15
min following acetic acid administration was counted and
recorded[10]. The percentage of inhibition was calculated
using the following formula:
(Writhing Number control－Writhing Number drug)/
Writhing Number control ×100%
2.5 Data analysis
Data were expressed as means±s ( ±x s ). Statistical
comparisons of the results were made using analysis of
ZHAO Chuan, et al. /Chinese Journal of Natural Medicines 2009, 7(4): 307−311
2009 年 7 月 第 7 卷 第 4 期 Chin J Nat Med July 2009 Vol. 7 No. 4 309

Table 1 Anti-inflammatory effects of HI and IJ injection on carrageenan-induced paw edema of rats ( ±x s , n=10)
Paw edema (mL)
Groups Dose (mL·kg−1)
0.5 h 1 h 2 h 4 h 6 h
Control 10 23.1±6.8 28.9±9.2 51.6±13.0 56.4±17.3 62.1±20.3
Diclofenac sodium 10 12.0±3.2*** 17.6±5.9** 23.4±6.9*** 27.9±5.4*** 37.8±11.3**
IJ injection 3.75 20.3±6.7 21.6±0.46* 31.1±9.7*** 44.8±10.3 53.1±14.3
7.5 19.0±5.4 26.2±4.1 34.5±4.2*** 39.3±6.1** 43.2±13.7*
15 13.1±2.7*** 18.5±2.9** 23.2±6.0*** 27.3±8.1*** 39.7±6.1**
HI injection 3.75 17.5±6.3 27.7±8.4 39.3±6.1* 45.6±8.0 49.8±11.0
7.5 15.9±7.1* 23.9±6.5 33.0±8.6** 38.8±9.1* 45.8±10.8*
15 10.8±4.8*** 20.7±5.5* 28.2±8.3*** 32.3±8.3*** 39.1±9.4**
*P<0.05, **P<0.01, ***P<0.001 vs control group; HI: extract isolated from the root bark of Illicium lanceolatum A. C. Smith; IJ: extract
isolated from Illicium Jiadifengpi B.N. Chang

variance (ANOVA) test. Significant difference was shown
as *P<0.05, **P <0.01 or ***P <0.001.
3 Results
3.1 Effects of injections on paw edema test
The hypodermic administration of both injections in-
hibited significantly, in a dose-dependent manner, the in-
crease of rat paw induced by carrageenin. Both of HI and IJ
injection were able to reduce the edema formation at 2 h, 4
h and 6 h, when administered at 7.5 mL·kg−1 (P<0.05,
P<0.01 and P<0.001) and 15 mL·kg−1 (P<0.01 and
P<0.001). All doses of HI and IJ injections significantly
inhibited the carrageenan edema at 2h. According to the
statistical data, no significant difference was observed be-
tween the two injections (Table 1).
3.2 Effects of injections on ear edema test
The results obtained from xylene-induced mice ear
edema test are demonstrated (Table 2). It was observed that
both injections showed significant inhibition of the xy-
lene-induced mice ear edema in a dose-dependent manner
as compared to the negative, with values ranging from 28 to
56%. As a positive control, diclofenac sodium (20 mL·kg−1)
gave rise to an inhibition of 42%. The inhibition percentage
of HI on xylene-induced ear edema in mice was equal to IJ
injection at 30mL·kg−1 and higher than IJ injection at 7.5
L·kg−1and 15 L·kg−1. According to the statistical data, no
significant difference was observed between the two injec-
tions.
3.3 Effects of injections on acid-induced vascular perme-
ability in mice
According to the statistical data (Table 3), both injec-
tions significantly inhibited acetic acid-induced vascular
permeability in mice in a dose-dependent manner (P<0.001)
and no significant difference was observed between the two
injections.
3.4 Effects of injections on hot-plate test
All other reagents showed significant increase of pain
threshold of mice in a dose-dependent manner (Table 4).
The analgesic effect of HI injection was higher than that of
diclofenac sodium (20 mL·kg−1) at the dose of 15 mL·kg−1
at 30 min and 60 min (P<0.05) and was lower than IJ injec-
tion at all doses in all times after injection. According to the
statistical data, no significant difference was observed be-
tween the two injections.

Table 2 Anti-inflammatory effects of HI and IJ injection
on xylene-induced ear edema of mice ( ±x s , n=10)
Groups Dose (mL·kg−1)
Ear edema
(mg)
Inhibition of
edema (%)
Control 20 7.7±1.33 -
Diclofenac sodium 20 4.5±0.68*** 42
IJ injection 7.5 5.5±1.32** 28
15 4.5±0.98*** 42
30 3.4±0.99*** 56
HI injection 7.5 5.3±1.27*** 31
15 4.1±1.25*** 47
30 3.4±1.04*** 56
*P<0.05, **P<0.01, ***P<0.001 vs control group; HI: extract
isolated from the root bark of Illicium lanceolatum A. C. Smith; IJ:
extract isolated from Illicium Jiadifengpi B.N. Chang

Table 3 Effects of HI and IJ injection on acetic acid-indu-
ced vascular permeability of mice ( ±x s , n=10)
Groups Dose (mL·kg−1) OD
Control 20 0.188±0.025
Diclofenac sodium 20 0.048±0.018***
IJ injection 7.5 0.079±0.023***
15 0.062±0.019***
30 0.050±0.015***
HI injection 7.5 0.084±0.022***
15 0.063±0.018***
30 0.053±0.014***
*P<0.05, **P<0.01, ***P<0.001 vs control group; HI: extract
isolated from the root bark of Illicium lanceolatum A. C. Smith; IJ:
extract isolated from Illicium Jiadifengpi B.N. Chang
ZHAO Chuan, et al. /Chinese Journal of Natural Medicines 2009, 7(4): 307−311
310 Chin J Nat Med July 2009 Vol. 7 No.4 2009 年 7 月 第 7 卷 第 4 期

Table 4 Analgesic effects of HI and IJ injection in hot-plate test of mice ( ±x s , n=10)
latency of pain response (s)
Groups Dose (mL·kg−1)
30 min 60 min 90 min
Control 20 13.23±1.86 13.55±2.93 13.48±3.48
Diclofenac sodium 20 14.91±1.76* 18.77±2.91*** 26.57±2.49***
7.5 15.49±2.54* 17.66±2.60* 21.91±3.92***
15 16.98±3.06** 21.95±4.52*** 27.64±3.61*** IJ injection
30 19.86±5.85** 25.59±6.40*** 32.64±8.94***
7.5 14.71±1.88* 16.62±2.18*** 20.40±1.90***
15 15.03±2.17* 19.11±5.34* 25.41±6.71*** HI injection
30 15.97±3.10* 21.95±6.89** 28.30±6.62***
*P<0.05, **P<0.01, ***P<0.001 vs control group; HI: extract isolated from the root bark of Illicium lanceolatum A. C. Smith; IJ: extract
isolated from Illicium Jiadifengpi B.N. Chang

3.5 Effects of injections on abdominal constriction test
As shown in Table 6, both HI and IJ injections (7.5-30
mL·kg−1) showed significant inhibition of peripheral pain in
a dose-dependent manner. Statistically, both injections at all
doses produced significant inhibition for all observed times
(P<0.001) and no significant difference was observed be-
tween the two injections. The analgesic effect of the injec-
tions at the dose of 30 mL·kg−1 showed higher pain inhibi-
tion intensity than that of diclofenac sodium at 20 mL·kg−1.

Table 5 Analgesic effects of HI and IJ injection in ab-
dominal constriction test of mice ( ±x s , n=10)
Groups Dose (mL·kg−1)
Number of
writhing
Inhibiting
rate (%)
Control 20 29.0±4.08 -
Diclofenac sodium 20 13.0±3.46*** 55.2
IJ injection 7.5 17.4±5.62*** 40.0
15 15.2±3.01*** 47.6
30 9.8±2.30 *** 66.2
HI injection 7.5 17.7±4.55*** 38.9
15 16.5±3.87*** 43.1
30 9.7±1.70 *** 66.5
*P<0.05, **P<0.01, ***P<0.001 vs control group; HI: extract
isolated from the root bark of Illicium lanceolatum A. C. Smith; IJ:
extract isolated from Illicium Jiadifengpi B.N. Chang
4 Discussion
In the present study, anti-inflammatory and analgesic
effects of two extracts isolated from Illicium angustise-
palum A. C. Smith and Illicium Jiadifengpi B.N. Chang,
respectively, were evaluated and compared using in vivo
animal models. Diclofenac sodium, a common
anti-inflammatory drug, which reduced inflammation, swel-
ling and arthritic pain by inhibiting prostaglandins synthesis
and/or production[11,12], was used as positive drug in this
experimental animal model[13].
Anti-inflammatory effects were evaluated using car-
rageenan-induced rat paw edema and xylene-induced mice
ear edema models. Carrageenan-induced edema is a classi-
cal pharmacological model in the study of an-
ti-inflammatory drugs[14]. The early phase (1.5-4 h) was due
to histamine and serotonin release from perivenular mast
cells and activation of plasma proteins. The later phase
(4.5-6 h) was attributed to the activation of kinin system
and bradykinin release. The results showed that both ex-
tracts significantly inhibited carrageenan-induced paw
edema with more potent effects at 2 and 4 h (the early
phase), which may be related to the mediation of histamine
and serotonin release. The xylene-induced mouse ear edema
is commonly used for screening anti-inflammatory agents
with certain advantage for natural product testing[15]. The
model of acetic acid-elevated vascular permeability, which
in some ways mimics clinical features, was also employed
in this study to evaluate anti-inflammatory effects of two
extracts in vivo.
Analgesic effects were evaluated using thermal
(hot-plate test) and chemical (acetic acid-induced writhing)
irritation model. The hot-plate test was a central an-
ti-nociceptive pain test and commonly used for assaying
narcotic-like analgesics[16]. Acetic acid-induced writhing
test was commonly regarded as a peripheral nociceptive
pain model[17]. The results showed that both extracts sig-
nificantly increased the latency of pain response and de-
creased the number of writhing, it suggesting that extracts
may act through both peripheral and the central nervous
system to inhibit pain responses.
In this study, both extracts isolated from Illicium an-
gustisepalum A. C. Smith and Illicium Jiadifengpi B.N.
Chang, respectively, displayed marked and identical an-
ti-inflammatory and analgesic effects without significant
difference according to the statistical data. The results sup-
plied pharmacological supports to the reported folkloric
uses of the root bark of Illicium Jiadifengpi B.N. Chang in
the control of painful and inflammatory conditions. Illicium
ZHAO Chuan, et al. /Chinese Journal of Natural Medicines 2009, 7(4): 307−311
2009 年 7 月 第 7 卷 第 4 期 Chin J Nat Med July 2009 Vol. 7 No. 4 311

Jiadifengpi B.N. Chang was possibly used as a substitute of
Illicium angustisepalum A. C. Smith. However, further
pharmacological and toxicological experiments are neces-
sary to evaluate the effectiveness and safety of Illicium
Jiadifengpi B.N. Chang as a substitute for Illicium angusti-
sepalum A. C. Smith in clinical use.
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两种八角属木兰科植物提取物抗炎镇痛作用的比较
赵 氚 1, 何 玲 1*, 张陆勇 2, 单 牧 1
1 中国药科大学药理教研室, 南京 210009;
2 中国药科大学新药筛选中心, 南京 210009
【摘 要】 目的：由于狭叶茴香属于我国的濒危野生物种之一, 寻找狭叶茴香的理想替代药材非常必要。本研究的
目的是比较狭叶茴香和假地枫皮这两种八角属植物的根皮提取物注射液的抗炎镇痛作用。方法：本实验采用大鼠角叉菜
胶足跖肿胀模型, 小鼠耳廓二甲苯致炎模型以及醋酸引起的小鼠腹腔毛细血管通透性模型评价这两种注射液的抗炎活性。
小鼠热板致痛模型和小鼠醋酸致痛模型评价这两种注射液的镇痛活性。结果：这两种注射液均表现出显著的抗炎镇痛作
用。实验结果中没有出现显著性差异。结论：假地枫皮的根皮提取物注射液的抗炎镇痛作用与狭叶茴香的相当。实验结
果显示, 假地枫皮有可能成为狭叶茴香的理想替代药材。
【关键词】 木兰科; 狭叶茴香; 假地枫皮; 抗炎; 镇痛

【基金项目】 财政部 2007 年度公益性行业科研专项：中医药行业专项(200707008); 江苏省药效平台资助项目 (BM2005103)