全 文 ：8 位被脂肪氨基对称地取代, 显示细胞毒作用最强,
抗癌活性明显提高, 至于它们抗M DR 肿瘤细胞作
用是否也与它们的这种结构有关还有待继续研究。
RO S 是指氧的某些代谢产物和一些反应的含
氧产物。生理状态下, 机体产生的自由基与抗氧化防
御系统处于相对平衡。在各种病理因子作用下, 机体
产生大量自由基, 或机体抗氧化防御系统受到破坏,
造成细胞结构和功能的破坏。本实验结果表明 4 种
药物分别与两种细胞共同培养 12、24、48 h, 12 h 即
能引起细胞内 RO S 明显增加, 24 h 时细胞内 RO S
的增加达到最大, 曲线明显右移, 48 h 细胞内 RO S
不再显著性增加, 甚至个别出现曲线微弱左移 (图
略)。同时, 用D iOC 6检测到 4 种药物分别作用两种
细胞 12、24 h 后, 线粒体跨膜电位 (∃7 m ) 稍有降
低, 至 48 h 后出现 ∃7 m 明显降低, 这与Co rtassa [9 ]
等的实验结果相吻合, 表明 RO S 的增加可直接或
间接损伤线粒体膜, 造成膜电位下降, 一些学者的研
究结果也支持这一结论[9, 10 ]。
研究发现 RO S 可能作为信号分子介导了细胞
对促凋亡信号的反应。因此, 推测这 4 种大黄素蒽醌
衍生物抑制 KB 和 KBv200 细胞的增殖, 可能与其
通过线粒体途径诱导细胞凋亡有关。因为它们均能
增加细胞内 RO S, 当 RO S 增加到一定程度, 即引起
细胞脂质过氧化, 从而干扰细胞线粒体的功能使得
线粒体M PT P 开放, 不仅导致跨膜电位崩溃, 也使
细胞色素 C 外漏, 最终启动 Caspase 的级联活化从
而引起细胞凋亡, 这就是所谓的线粒体依赖性凋亡
通路。这 4 种大黄素蒽醌衍生物是否通过该通路发
挥抗肿瘤作用, 相关的研究正在进行中。此外, 它们
是在大黄素母核结构基础上经过不同基团修饰合成
的, 它们对M DR 细胞无抗药性, 因此对它们构效关
系及作用机制关系的进一步研究, 有可能为开发较
大黄素作用更强的抗M DR 肿瘤的药物提供线索。
References:
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m an hepatocellu lar carcinom a Bel7402 cell line [J ]1 Cancer
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[ 7 ]　Kum ar A , D haw an S, A ggerw al B B1 Emodin (32m ethyl21,
6, 82trihydroxyan2th rau inone) inh ib its TN F2induced N F2ϑB
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[ 8 ]　Cheng Y W , Kang J J1 Emodin2induced m uscles con traction
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p hy s J , 2004, 87 (3) : 2060220731
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[ 11 ]　D ing W X, Shen H M , O ng C N 1 C ritical ro le of reactive
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An ti- inf lammatory activ ity of ethanol extracts
from root of D ap hne genkwa
ZH EN G W ei2fa1, W AN G L i2, SH I Feng1Ξ
(1. Key L abo rato ry fo r B io techno logy on M edicinal P lan ts of J iangsu P rovince, Xuzhou N o rm al U niversity, Xuzhou
221116, Ch ina; 2. Schoo l of L ife Science, Yangzhou U niversity, Yangzhou 225009, Ch ina)
　　Abstract: Object　To elucidate the an t i2inf lamm ato ry act ivity of the ethano l ex tracts from roo ts of
·2621· 中草药　Ch inese T radit ional and H erbal D rugs　第 35 卷第 11 期 2004 年 11 月
Ξ 收稿日期: 2004202206
基金项目: 教育部科学技术研究重点项目 (03049) ; 江苏省教育厅自然科学基金重点项目 (02KJA 360002) ; 江苏省药用植物生物技术
重点实验室开放基金 (KJS02114)
作者简介: 郑维发 (1962—) , 男, 安徽南陵人, 理学博士, 教授, 研究方向为天然产物化学及其药理毒理学。
T elöFax: (0516) 3403179　E2m ail: yyzw @xznu. edu. cn
D ap hne g enkw a Sieb. et Zucc. (EERD ). M ethods　T he act ion on acu te inflamm ation w as evaluated by
the inh ib it ion of h istam ine2induced vascu lar perm eab ility, carrageen in2induced ra t paw edem a and phagocy2
to sis of ret icu lar endo thelia l system (R ES) in m ice. T he act ion on ch ron ic inflamm ation w as exam ined u s2
ing adjuvan t2induced arth rit is and co t ton pellet2induced granu lom a. Results　EERD p roduced eviden t inh i2
b it ion of vascu lar perm eab ility and ra t paw edem a at a do se of 40 m gökg and enhanced phagocyto sis of R ES
in m ice at a do se of 30 m gökg. D aily trea tm en t a t a do se of 30 m gökg exh ib ited sign if ican t inh ib it ion of
granu lom a and adjuvan t2induced po lyarth rit is. EERD also inh ib ited the p roduct ion of M DA and PGE2,
NO , TN F2Α and IL 21Β, enhanced the act ivity of SOD and CA T and reduced the act ivity of iNO S.
Conclusion　EERD acts as an t i2inf lamm ato ry agen t by m echan ism s that invo lve the inh ib it ion of lip id per2
ox idat ion and the release of m edia to rs, the enhancem en t of act ivity of SOD and CA T , and the reduct ion in
act ivity of iNO S as w ell as the p romo tion of phagocyto sis of R ES.
Key words: ethano l ex tracts from roo ts of D ap hne g enkw a Sieb. et Zucc. (EERD ) ; an t i2inf lamm ato ry
act ivity; lip id perox idat ion react ion
芫花根乙醇提取物的抗炎活性
郑维发1, 王　莉2, 石　枫1
(11 徐州师范大学 江苏省药用植物生物技术重点实验室, 江苏 徐州　221116; 21 扬州大学生命科学院, 江苏 扬州　225009)
摘　要: 目的　阐明芫花根乙醇提取物 (EERD ) 的抗炎活性。方法　EERD 对急性炎症的抑制作用采用组胺诱导
的小鼠毛细血管通透性增加、角叉菜胶诱导的大鼠足肿胀以及小鼠网状内皮系统 (R ES) 对异物的吞噬作用进行
分析。对慢性炎症的抑制作用以福氏完全佐剂诱导的多发性关节炎和棉球诱导的大鼠肉芽肿进行分析。结果　
EERD 在剂量为 40 m gökg 时能明显地抑制小鼠毛细血管通透性增加和抑制大鼠足肿胀; 在剂量为 30 m gökg 时
对 R ES 的吞噬能力有提升作用、对大鼠肉芽肿和多发性关节炎表现出显著抑制作用。EERD 也能抑制丙二醛
(M DA )、前列腺素 E2 (PGE 2)、一氧化氮 (NO )、肿瘤坏死因子2Α (TN F2Α) 和 白细胞介素21Β ( IL 21Β) 的形成, 增强
超氧化物歧化酶 (SOD ) 和过氧化氢酶 (CA T ) 的活力并钝化诱导型氮氧化物合酶 ( iNO S) 的活性。结论　EERD
的抗炎活性主要是通过抑制脂质过氧化反应和炎症介质的释放、增强 SOD 和 CA T 的活力、钝化 iNO S 的活性以
及提升 R ES 的吞噬作用实现的。
关键词: 芫花根乙醇提取物; 抗炎活性; 脂质过氧化反应
中图分类号: R 286175　　　文献标识码: A 　　　文章编号: 0253 2670 (2004) 11 1262 08
1　 In troduction
　　D ap hne g enkw a Sieb. et. Zucc. (T hym elaea2
ceae ) is w idely dist ribu ted in sou thern Ch ina as
w ell as the region s along the Yellow R iver [1 ]. Fo r
years, th is species has been u sed as the herbal
rem edy fo r t rea t ing ch ron ic b ronch it is, hepat it is,
and arth rit is etc[1 ]. A num ber of studies have dis2
clo sed that the roo ts of D. g enkw a po ssess an t i2in2
f lamm ato ry, analgesic, an t iconvu lsan t and an t i2ir2
rita te act ivity and p resen t eviden t eff icacy in relie2
ving cough and asthm a and exh ib it rem arkab le bac2
terio sta t ic act ivity again st pneumococcu s and op2
po rtun ist ic derm atophytes [1 ]. T he roo ts of the
species have also been u sed in Ch ina as the m ajo r
componen t in the p rescrip t ion fo r t rea t ing rheum a2
to id arth rit is[2 ]. D esp ite its m edicinal impo rtance,
how ever, th is species has no t yet been sub jected to
system at ic pharm aco logica l an t i2inf lamm ato ry
evaluat ion to sub stan t ia te its app lica t ion on thera2
peu t ic pu rpo ses. M o reover, the m echan ism s fo r
an t i2inf lamm ato ry effect of the species have no t
been clarif ied as yet. In an at temp t to fu rther con2
f irm the an t i2inf lamm ato ry act ivity of the roo ts of
D. g enkw a and its po ssib le m echan ism s of act ion,
the study on inh ib it ion of acu te and ch ron ic infla2
mm ation by ethano l ex tracts from roo ts of D . g en2
kw a (EERD ) w as perfo rm ed. T h is paper described
in v ivo eff icacy of the ex tract in inh ib it ing acu te
and ch ron ic inflamm ation of an im als and gave po s2
sib le exp lanat ion on m echan ism s of act ion.
2　M a ter ia ls and m ethods
2. 1　M ateria ls: T he roo ts of D. g enkw a w ere co2
llected from sou thern moun ta inou s region of A nhu i
P rovince, Ch ina, in N ovem ber 2002, and au then t i2
·3621·中草药　Ch inese T radit ional and H erbal D rugs　第 35 卷第 11 期 2004 年 11 月
cated by P rof. YE D ing2jiang, from Facu lty of
Pharm acy, N an jing U n iversity of T radit ional
Ch inese M edicine, Ch ina. A voucher specim en
TCM 978 has been p reserved in the H erbarium fo r
Facu lty of Pharm acy, N an jing U n iversity of T radi2
t ional Ch inese M edicine, Ch ina.
2. 2　A n im als: W istar ra ts w eigh ing 180—220 g
w ere u sed fo r the assay of effects on ch ron ic and
sub2acu te inflamm ation and KM m ice w eigh ing
18—22 g on acu te inflamm ation. T he an im als w ere
hou sed at (23±1) ℃, w ith rela t ive hum idity of
(55±10) % , 12ö12 h ligh tödark cycle and fed w ith
standard pellet d iet and w ater ad libitum . T he
an im als w ere cared hum an ly acco rd ing to the
standards fo r labo ra to ry an im als estab lished by
Peop le’s R epub lic of Ch ina ( GB 14923294, GB
14922294, and GB öT 14925294).
2. 3　M ethods
2. 3. 1　P repara t ion of ethano l ex tracts: T he w ell2
pu lverized, aeria l dried roo ts of D. g enkw a (5 kg)
w ere ex tracted w ith 32fo ld vo lum e of 95% E tOH
fo r seven days at room temperatu re and fu rther
concen tra ted in vacuum to affo rd 495 g ex tracts
(accoun ts fo r 9. 9% of the to ta l m ateria l).
2. 3. 2　V ascu lar perm eab ility of m ice: M ice w ere
random ly classif ied in to th ree test group s and tw o
con tro l group s (n = 12). T he test group s w ere ig
adm in istered EERD em u lsion p repared in Tow een2
80 (ShanghaiN o. 2 R eagen t P lan t, Ch ina) a t do ses
of 10, 30, and 50 m gökg, respect ively. T he con2
t ro l group s ig received saline (b lank con tro l) and
p redn isone aceta te (PR E ) (Sigm a, po sit ive con2
t ro l) a t a do se of 30 m gökg. O ne hou r after EERD
treatm en t, the test m ice w ere sc syringed w ith
h istam ine (Sigm a) at a do se of 1 m gömL ×200 ΛL
fo llow ed by iv in ject ion of 1% Evan s B lue (Sigm a)
at a do se of 100 m gökg. Tw en ty m inu tes la ter, the
t inged area of sk in and its con ten t of Evan s B lue
w ere assayed as p reviou sly described [3 ].
2. 3. 3　Carrageen in2induced ra t paw edem a: W is2
ta r ra ts w ere random ly classif ied in to five test
group s and tw o con tro l group s (n = 10). T he test
group s w ere ig trea ted by EERD at do ses of 10,
20, 30, 40, and 50 m gökg, respect ively, 30 m in
p rio r to in ject ing of carrageen in (Sigm a ) ( 1% ×
0. 1 mL ) in to righ t h ind paw of test ra ts. O ne hou r
after inflamm ation, paw edem a of the ra ts w as
detected every hou r fo r six con secu t ive hou rs as
p reviou sly described [4 ]. T he o ther tw o group s
receiving indom ethacin ( ID T ) at a do se of 3. 6 m gö
kg and saline w ere u sed as po sit ive and b lank con2
t ro l, respect ively.
2. 3. 4　Con ten ts of M DA and m edia to rs, and ac2
t ivity of enzym es invo lved in carrageen in2induced
inflamm ation: W istar ra ts w ere random ly divided
in to five test group s and tw o con tro l group s (n =
10). T he test group s w ere ig m edicated by EERD
at do ses of 10, 20, 30, 40, and 50 m gökg, respec2
t ively, 30 m in p rio r to in ject ing 0. 1 mL of 1% car2
rageen in to the h ind paw of test ra ts. Fou r hou rs
after expo su re to carrageen in, the homogenate of
edem a paw of the ra ts w as p repared fo r determ i2
n ing the con ten ts of p ro stag landin E 2 ( PGE2 ) ,
m adonadia ldehyde (M DA ) , and NO , and the ac2
t ivity of SOD , CA T as w ell as iNO S acco rd ing to
the p rocedu res described in specif ica t ion s of re2
agen t k its (N an jing J iancheng B io techno logy In s2
t itu te). Con tro l group s w ere trea ted w ith ID T at a
do se of 3. 6 m gökg and saline, respect ively. L evels
of PGE 2, M DA , and NO w ere exp ressed as nmo lö
m g, w h ile the levels of SOD , CA T , and iNO S
w ere p resen ted in nU öm g.
2. 3. 5 　 F reud’s comp lete adjuvan t2induced paw
edem a: W istar ra ts w ere random ly classif ied in to
five test group s and tw o con tro l group s (n = 10).
T he test group s w ere ig m edicated by EERD at
do ses of 10, 20, 30, 40, and 50 m gökg, respec2
t ively, one hou r p rio r to in ject ing 0. 1 mL F reund’s
comp lete adjuvan t (Sigm a) to the righ t h ind paw
of test ra ts. T he increase in paw edem a w as
reco rded every o ther day fo r 22 con secu t ive days to
calcu la te edem a rate as p reviou sly illu st ra ted [4 ].
T he tw o con tro l group s w ere received PR E at a
do se of 30 m gökg (po sit ive con tro l) and saline
(b lank con tro l) , respect ively.
2. 3. 6　Fo rm at ion of M DA , and m edia to rs, and
enzym es invo lved in adjuvan t2induced inflamm a2
t ion: W istar ra ts w ere random ly separa ted in to five
·4621· 中草药　Ch inese T radit ional and H erbal D rugs　第 35 卷第 11 期 2004 年 11 月
test group s and tw o con tro l group s (n= 10) , each
receiving 0. 1 mL F reud’s comp lete adjuvat on their
h ind paw. T he five test group s of AA rats w ere ig
trea ted by EERD at do ses of 10, 20, 30, 40, and
50 m gökg, respect ively, fo r 19 con secu t ive days.
O ne hou r after the last m edicat ion, the inflam ed
paw of the ra ts w as homogenated w ith PBS in ice
bath fo llow ed by cen trifugat ion to p roduce super2
natan ts. T he supernatan t of homogenate w as as2
sayed fo r the con ten t of IL 21Β, TN F2Α (T P I Inc.
U SA ) , M DA , PGE2, and NO , and the act ivity of
SOD , CA T , and iNO S acco rd ing to the p rocedu res
described in the specif ica t ion s of reagen t k its. T he
tw o con tro l group s of AA rats received ID T at a
do se of 3. 6 m gökg (po sit ive) and saline (b lank).
L evels of PGE2, M DA , and NO w ere deno ted in
mo löm g, and levels of SOD , CA T , and iNO S in
nU öm g, w h ile levels of IL 21Β, TN F2Α in pgömL.
2. 3. 7　Co tton pellet2induced granu lom a: W istar
ra ts w ere random ly divided in to five test group s
and tw o con tro l group s (n = 10). T he test group s
w ere ig adm in istered EERD at do ses of 10, 20, 30,
40, and 50 m gökg fo r seven con secu t ive days, res2
pect ively, th ree days p rio r to the opera t ing fo r the
fo rm at ion of granu lom a developed by co t ton pellet
acco rd ing to the m ethod described by H ajare [5 ].
T he inh ib it ion of the grow th of granu lom a w as as2
sayed u sing the tw o con tro l group s receiving ID T
at a do se of 3. 6 m gökg and saline, respect ively.
2. 3. 8　Phagocyto sis of R ES in m ice: M ice (♀ö®) w ere random ly separa ted in to five test group s
and tw o con tro l group s (n = 20). T he test group s
w ere o ra lly t rea ted by EERD at do ses of 10, 20,
30, 40, and 50 m gökg fo r f ive con secu t ive days.
O ne hou r after the last m edicat ion, the test m ice
w ere iv syringed w ith 1% congo2red fo r determ in2
ing the expu rgat ion by ret icu lar endo thelia l system
(R ES ) as p reviou sly described [1 ]. T he con tro l
group s w ere ig trea ted w ith ID T at a do se of 3. 6
m gökg and saline, respect ively.
2. 3. 9　Stat ist ica l analysis: T he data acqu ired in
the experim en t w ere p rocessed u sing Studen t2N ew 2
m an2Keu ls test softw are. T he resu lts w ere indica t2
ed as x ±s.
3　Results
3. 1　Effect on vascu lar perm eab ility in m ice: In2
f lamm ation is concom itan t w ith the increase of vas2
cu lar perm eab ility. EERD in u sed do ses exh ib ited
rem arkab le inh ib ito ry effect on h istam ine2induced
increase of vascu lar perm eab ility. A t a do se of 10
m gökg, EERD p roduced eviden t reduct ion in
Evan s B lue perm eated from cap illary b lood vessel
by 52. 5% and indica ted do se2dependen t inh ib it ion
of up to 68. 5%. In con trast to the m ice trea ted
w ith sa line, the t inged area by Evan s B lue w as also
reduced sign if ican t ly and do se2dependen t ly. By
comparison, PR E show ed sim ilar effect in inh ib it2
ing vascu lar perm eab ility to EERD (T ab le 1).
Table 1　Effect of EERD on vascular permeabil ity in m ice
induced by h istam ine (x±s, n= 12)
Group s
Do seö(m g·kg- 1) T inged areaömm 2 Evens B lueö(m g·mL - 1) Inh ib itionrateö%
N o rm al - 　0102± 0100 0100±0100 -
Saline - 450123±35117△ 0162±0108△ -
PR E 30 288113±291333 3 3 0129±01023 59105
EERD 10 319187±221893 3 0130±01013 52175
30 294119±211973 3 3 0129±01003 55190
50 128176±171473 3 3 0127±01013 68150
　　△P < 0101 vs no rm al group
　　3 P < 0105　3 3 P < 0101　3 3 3 P < 01001 vs saline group
3. 2　Effect on carrageen in2induced paw edem a of
ra ts: T heo ret ica lly, carrageen in2induced paw ede2
m a belongs to sub2acu te inflamm ation. T he paw
edem a of the ra ts t rea ted by saline ro se from
24. 8% at one hou r to 55. 1% at th ree hou rs fo l2
low ing the expo su re to carrageen in and began to
scale dow n from 33. 9% at fou r hou rs to 30. 8% at
six hou rs. In con trast, the paw edem a of the ra ts
t rea ted by EERD at u sed do ses did no t exceed
18. 1% at one hou r and 26. 7% at th ree hou rs. T he
paw edem a of the ra ts w as decreased to less than
20. 4% beginn ing from fou r hou rs. In comparison,
the EERD at u sed do ses and ID T at do se of 3. 6
m gökg indica ted no sign if ican t d ifference in inh ib i2
t ion of carrageen in2induced paw edem a (F ig. 1).
3. 3　Effects on the genera t ion of M DA , and m e2
dia to rs, and act ivity of invo lved enzym es in car2
rageen in2induced infamm ation: EERD in u sed do s2
es p roduced po sit ive reduct ion in genera t ion of
M DA , PGE2 , and NO . T he inh ib it ion of M DA
·5621·中草药　Ch inese T radit ional and H erbal D rugs　第 35 卷第 11 期 2004 年 11 月
F ig. 1　Effect of EERD on carrageen in- induced
paw edema of rats (x ±s, n = 10)
genera t ion w as ob served at a do se of 10 m gökg
(P < 01001) and in ten sif ied in respon se to the do se
increase w ith in 30 m gökg. Fu rther increase in do se
trea tm en t ( i. e. 40 m gökg) , how ever, resu lted in
reduced level of inh ib it ion. O bviou s inh ib it ion on
the release of PGE 2 and NO w as detected at a do se
of 1 0m g ökg ( P < 0 1 0 5 ) and fu rther enhanced in respon se to the increased do se2t rea tm en t below 40m gökg. A t a do se of 50 m gökg, nevertheless,EERD p resen ted reduced inh ib it ion on fo rm at ion ofPGE 2 and NO (T ab le 2).　　W ith reference to the effects on enzym es in2vo lved in inflamm ation, EERD in u sed do ses re2vealed p rom inen t increase in the act ivity of SODand CA T. Below the do se of 40 m gökg, EERDp roduced rem arkab le enhancem en t on the act ivityof SOD , w h ile a t a do se of 50 m gökg, EERD gen2era ted less enhancem en t in the act ivity of SOD.EERD enhanced the act ivity of CA T at a do se of 30m gökg (T ab le 3). O n the o ther hand, EERD at ado se of 20 m gökg indica ted po sit ive inh ib it ion onthe act ivity of iNO S and in ten sif ied fo llow ing theincrease of do se trea tm en t reach ing its m ax im al in2h ib it ion at a do se of 40 m gökg. Fu rther increase indo se trea tm en t, how ever, EERD p roduced lesslevel of inh ib it ion on the iNO S act ivity (T ab le 3).3. 4 Effects on adjuvan t2induced paw edem a of
Table 2　Effect of EERD on carrageen in- induced generation of MDA and mediators (x±s, n= 10)
Group s Do seö(m g·kg- 1) M DA ö(nmo l·m g- 1) PGE2ö(nmo l·m g- 1) NO ö(nmo l·m g- 1)
N o rm al 　　　　- 01230±0104 01278±0111 1139±0159
Saline 　　　　- 01673±0109△△ 01683±0112△△ 1177±0158△
ID T 　　　　316 01263±01093 3 3 01459±01103 1113±01493 3
EERD 10 01462±01043 01367±01073 3 1149±01513
20 01383±01063 3 01375±01113 3 1131±01433
30 01237±01083 3 3 01329±01063 3 0185±01383 3 3
40 01449±01093 01289±01083 3 0179±01223 3 3
50 01633±0107 01539±0109 0193±01393 3 3
　　　△P < 0105　△△P < 0101 vs no rm al group; 　3 P < 0105　3 3 P < 0101　3 3 3 P < 01001 vs saline group
Table 3　Effects of EERD on activ ity of enzymes involved in carrageen in- induced inf lammation (x±s, n= 10)
Group s Do seö(m g·kg- 1) SOD ö(nU ·m g- 1) CA Tö(nU ·m g- 1) iNO Sö(nU ·m g- 1)
N o rm al 　　　- 32153±1167 0189±0122 01152±0102
Saline 　　　- 25199±1107△ 0158±0116△ 01292±0105△△
ID T 　　　316 30155±31023 3 3 0197±01253 3 3 01156±01033 3
EERD 10 30181±51143 3 3 0156±0113 01263±0101
20 30165±51123 3 3 0158±0113 01232±01013
30 31133±11233 3 3 0167±01123 3 01213±01023
40 30120±41283 3 3 0179±01133 3 01167±01023 3
50 28144±31713 3 3 0186±01083 3 3 01186±01033 3
　　　△P < 0105　△△P < 0101 vs no rm al group; 　3 P < 0105　3 3 P < 0101　3 3 3 P < 01001 vs saline group
AA rats: T he paw edem a of AA rats ro se from
23. 6% at the second day to fo llow ing the expo su re
to F reud’s comp lete adjuvan t. O n the eigh th day
(the end of la ten t phase) the edem a ro se ab rup t ly
from 40. 3% to 58. 9% and then fo llow ed by a
gradual decrease. EERD in u sed do ses show ed con2 siderab le inh ib ito ry effects on paw edem a of AArats. T he inh ib it ion by EERD began from thefou rth day and kep t ra ther steady inh ib it ive ra teun t il the tw en t ith day. Among the do ses u sed,daily t rea tm en t a t a do se of 30 m gökg rep resen tedthe m in im um edem a from the beginn ing of expo2
·6621· 中草药　Ch inese T radit ional and H erbal D rugs　第 35 卷第 11 期 2004 年 11 月
su re to F reud’s comp lete adjuvan t t ill the end of
the experim en t. T he inh ib it ion on paw edem a by
PR E, as show n in F ig. 2, w as sim ilar to that of
EERD at a do se of 10 m gökg.
F ig. 2　Effect of EERD on adjuvan t- induced
paw edema of rats (x ±s, n = 10)
3. 5 　 Effects on adjuvan t2induced fo rm at ion of
M DA and m edia to rs, and the act ivity of invo lved
enzym es in inflam ed paw of AA rats : T he release
of M DA and variou s m edia to rs including PGE2,
NO , IL 21Β, and TN F2Α is invo lved in the p rocess
of inflamm ation. EERD in u sed do ses p roduced ob2
viou s inh ib it ion of the genera t ion of M DA and m e2
dia to rs. T he inh ib it ion of M DA and PGE2 w as de2
tected at a do se of 10 m gökg ( P < 01001) , en2
hanced at 20 m gökg and scaled dow n at a do se of
30 m gökg. T he inh ib it ion of NO , IL 21Β, and
TN F2Αw as ob served at a do se of 10 m gökg (P <
0105) and enhanced in respon se to the increase of
do se trea tm en t (T ab le 4). T he increase in act ivity
of SOD and CA T imp lica tes the enhanced inh ib i2
t ion of lip id perox idat ion. EERD enhanced the ac2
t ivity of SOD and CA T at a do se of 20 and 30 m gö
kg, respect ively (P < 0105). O n the o ther hand,
the reduct ion of NO is m edia ted by declin ing the
act ivity of iNO S. EERD w as so ob served to inh ib it
the act ivity of iNO S at a do se of 20 m gökg and in2
ten sif ied at 30 m gökg (T ab le 5).
Table 4　Effect of EERD on con ten ts of MDA and mediators in adjuvan t- induced inf lammation tissue (x±s, n= 10)
Group s
Do seö(m g·kg- 1) M DAö(nmo l·m g- 1) PGE2ö(nmo l·m g- 1) NOö(nmo l·m g- 1) IL 21Βö(pg·mL - 1) TN F2Αö(pg·mL - 1)
N o rm al 　　　- 0123±0104 01278±01109 1139±0159 20137±3105 186163±23140
Saline 　　　- 0148±0111△△ 01398±01110△ 3118±0195△△ 107139±15101△△△ 730151±63191△△△
ID T 　　　316 0130±01193 01292±010513 3 2101±01953 3 35123± 31383 3 3 299155±461323 3 3
EERD 10 0126±01093 3 01263±010503 3 2112±01783 3 51184± 31993 3 3 349194±581233 3 3
20 0121±01003 3 01252±010193 3 1148±01373 3 3 49119± 51723 3 3 283100±341593 3 3
30 0126±01013 3 01297±010283 1135±01683 3 3 38198± 31403 3 3 209100±611343 3 3
40 0129±01053 01321±01017 1144±01293 47156± 61823 3 248112±341893
50 0131±01043 01357±01043 1167±01373 59118± 91733 3 297137±561143
　　　△P < 0101　△△P < 01001　△△△P < 010001 vs no rm al group; 　3 P < 0105　3 3 P < 0101　3 3 3 P < 01001 vs saline group
Table 5　Effect of EERD on activ ity of involved enzymes in adjuvan t- induced inf lammation tissue (x±s, n= 10)
Group s Do seö(m g·kg- 1) SOD ö(nU ·m g- 1) CA Tö(nU ·m g- 1) iNO Sö(nU ·m g- 1)
N o rm al 　　　　- 32153±1167 0189±0122 0115±0102
Saline 　　　　- 21175±1114△△ 0141±0119△△ 0128±0109△
ID T 　　　　316 32138±21773 3 3 0169±01193 3 0120±01033 3
EERD 10 30187±21523 3 3 0181±01203 3 0121±01033 3
20 33157±31543 3 3 0186±01143 3 0117±01063 3
30 32119±21173 3 3 0192±0119 0115±01093 3
40 29179±31563 0179±0110 0122±01013 3
50 27184±41723 0170±0117 0126±01043
　　　△P < 0101　△△P < 01001 vs no rm al group; 　3 P < 0105　3 3 P < 0101　3 3 3 P < 01001 vs saline group
3. 6　Effects on developm en t of granu lom a in ra ts:
T he assay indica ted that EERD at a do se of 10 m gö
kg sligh t ly reduced the grow th of granu lom a by
24. 3% and gained reduct ion by 38. 56% and
54. 03% in respon se to the increase of do se trea t2
m en t from 20 to 30 m gökg, sym bo lizing its sign if i2 can t inh ib it ion of granu lom a fo rm at ion (P < 0101).Fu rther increase in do se trea tm en t (above 40 m gökg ) , how ever, p roduced less o r no inh ib it ion ofgranu lom a fo rm at ion (F ig. 3).3. 7　Effects on phagocyto sis of R ES: EERD indi2cated eviden t enhancem en t on phagocyto sis of R ES
·7621·中草药　Ch inese T radit ional and H erbal D rugs　第 35 卷第 11 期 2004 年 11 月
3 3 P < 0101　3 3 3 P < 01001 vs saline group
F ig. 3　Effect of EERD on cotton pellet
granuloma of rats (x±s, n= 10)
in test m ice. T he reduct ion of congo2red by phago2
cyto sis of R ES w as ob served at a do se of 20 m gökg
(P < 0105) and enhanced at a do se of 30 m gökg
(P < 0101). Fu rther increase in do se trea tm en t,
nevertheless, led to gradual decrease in phagocyt ic
act ivity of R ES. In comparison, the capacity fo r
enhancing phagocyto sis by ID T w as almo st the
sam e as that of EERD at a do se of 30 m gökg
(F ig. 4). 3 P < 0105　3 3 P < 0101 vs saline group
F ig. 4　Effect of EERD on phagocytosis
of RES in m ice (x±s, n= 10)
4　D iscussion
　　 Inflamm ation, w h ich invo lves bo th innate and
adap t ive imm une m echan ism s, is the respon se of
living t issue to cell in ju ry [4 ]. T he resu lts suggested
that EERD exh ib ited con siderab le inh ib it ion of h is2
tam ine2induced vascu lar perm eab ility, carrageen in2
induced ra t paw edem a, adjuvan t2induced granu lo2
m a, indica t ing that EERD w as an effect ive agen t
fo r the therapy of acu te and ch ron ic inflamm ation.
　　T he p rocess of inflamm ation is characterized
by the release of m edia to rs including h istam ine [3 ] ,
p ro stag landin s (PGs) , NO [6 ] , IL 2Β, and TN F2Α[7 ]
and by lip id perox idat ion that resu lts in the p ro2
duct ion of M DA [8 ]. H istam ine is released by m ast
cell th rough degranu la t ion and cau ses vasodila t ion
th rough com b in ing h istam ine recep to r on vascu lar
endo thelia l cells[9 ]. PGE2 is the m etabo lite of ar2
ach idon ic acid cata lyzed by cyclooxygenase du ring
the t issue in ju ry [10 ]. It has been repo rted that sm all
f luxes of NO p roduced by iNO S con tribu tes to the
imm une defen se again st invading m icroo rgan ism s
and tumo r cells. How ever, excessive genera t ion of
NO is associa ted w ith sep t ic shock, acu te and
ch ron ic inflamm ation, au to imm une diseases and
arterio sclero sis[11 ]. NO has also been evidenced to
be a st im u lan t rad ica l fo r the release of PGE2 [12 ].
IL 21 and TN F2Α are released m ain ly by act iva ted
monocytes and m acrophages du ring acu te inflam 2
m ation and term ed as p ro inflamm ato ry m edia2
to rs[13 ]. L ip id perox idat ion is t riggered by free
oxygen radica ls in a w ell2oxygenated environm en t
leading to the p roduct ion of M DA [14 ]. U nder phy2
sio logica l condit ion s, the p roduct ion of inflamm a2
to ry m edia to rs and free radica ls are abo lished by
SOD and CA T [15 ]. T he con siderab le inh ib it ion of
h istam ine2induced vascu lar perm eab ility and fo r2
m ation of PGE2 imp lies that EERD m ay act as an2
tagon ist of h istam ine recep to rs and the inh ib ito r of
cyclooxygenase. T he reduct ion in NO fo rm at ion
and inh ib it ion of act ivity of iNO S indica tes that
EERD is also the po ssib le iNO S inh ib ito r. T he re2
lease of IL 21 and TN F2Αby monocytes and m acro2
phages needs act iva t ion and in it ia t ion. O bviou s in2
h ib it ion on release of IL 21 and TN F2Α suggests
tha t p roper do se of EERD m igh t funct ion as the in2
h ib ito r fo r act iva t ion monocytes and m acrophages.
P rom inen t reduct ion by EERD in the con ten t of
M DA in inflam ed t issues fu rther confirm s the
quench ing and scavenging effect on free radica ls.
R ES is a part of ho st defen se system s again st in2
vading m icroo rgan ism s by phagocyto sis. Phagocy2
to sis is the p rocess by w h ich cells b ind and in ter2
nalize rela t ively large part icles con ta in ing dead cell
·8621· 中草药　Ch inese T radit ional and H erbal D rugs　第 35 卷第 11 期 2004 年 11 月
o r cell deb ris induced by comp lem en tary cascade
react ion [1 ] o r by m edia to rs th rough apop to sis[16 ].
T he strik ing enhancem en t fo r clearing Congo2red
in the b loodstream of m ice by EERD sym bo lizes its
op son ic effect on the phagoyto sis, w h ich is defi2
n itely help fu l in fa tcilita t ing rehab ilita t ion of in2
ju red t issues.
　 　 T herefo re, the an t i2inf lamm ato ry effect of
EERD w as realized, a t least in part, by inh ib it ing
lip id perox idat ion, release of m edia to rs and en2
hancing the act ivity of SOD and CA T as w ell as re2
ducing the act ivity of iNO S and po ten t ia t ing the
phagocyto sis of R ES.
References:
[ 1 ]　Zheng W F, Sh i F, W ang L , et a l. Chem ical componen ts of
non2po lar fractions from ethano l ex tract of radixes D ap hne
g enkw a and their inh ib it ion to acu te inflamm ation [J ]1 P har2
m a J Ch in PL A (解放军药学学报) , 2004, 20: 182211
[ 2 ]　Yang C L 1 T ox ic H erba l M ed icine (毒药本草) [M ]1 Bei2
jing: Ch ina T raditional Ch inese M edicine P ress, 19931
[ 3 ]　Yousif M H M , O riowo M A , Cherian A , et a l. H istam ine2
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[ 4 ]　Zheng W F, Chen C F1 A nti2inflamm ato ry activity of ch lo ro2
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1841
乌苏里藜芦碱对血小板聚集及凝血与出血时间的影响
李欣燕1, 韩国柱1Ξ , 张书文1, 吕　莉1, 杨静娴1, 赵伟杰2
(11 大连医科大学 药理教研室, 辽宁 大连　116027; 21 大连理工大学 制药工程系, 辽宁 大连　116024)
摘　要: 目的　研究乌苏里藜芦碱 (V nA ) 对大鼠的抗血小板作用及其对小鼠凝血时间和出血时间的影响。方法　
比浊法测定正常大鼠及血瘀模型大鼠血小板聚集百分率, 观察 V nA 抗血小板作用。毛细玻璃管法测定小鼠全血凝
血时间 (CT ) ; 比较等效抗凝剂量的 V nA 及肝素对小鼠尾出血时间 (BT ) 的影响。结果　V nA (45、30、15 Λgökg,
iv) 对AD P 诱发的大鼠血小板聚集有明显抑制作用, 且呈剂量依赖性。V nA (12. 5、25、50、100 Λgökg, ip ) 可明显
延长小鼠 CT 和BT , 等效抗凝剂量的V nA (4913 Λgökg, ip ) 所致BT 延长略低于肝素 (1125 m gökg, ip ) , 但无统
计学意义。结论　V nA 具有显著抗血小板作用, 能显著延长 CT , 对BT 的延长作用不超过肝素。
关键词: 乌苏里藜芦碱; 抗血小板作用; 凝血时间; 出血时间
中图分类号: R 286132　　　文献标识码: A 　　　文章编号: 0253 2670 (2004) 11 1269 04
·9621·中草药　Ch inese T radit ional and H erbal D rugs　第 35 卷第 11 期 2004 年 11 月
Ξ 收稿日期: 2004202211
基金项目: 辽宁省教委科研项目 (20272271)
作者简介: 韩国柱 (1943—) , 男, 江苏省泰州市人, 现为大连医科大学教授, 从事抗血栓药理学与药动学研究。
T el: (0411) 84720229　E2m ail: hgzhx2236@ sina. com. cn3 通讯作者