全 文 ：3个剂量组均能抑制 Neu 释放 O÷2 , 且呈剂量依赖
性,其抑制率分别为 10. 19%, 29. 96%, 41. 43%。
2. 4　对灌洗液中 Neu 内 cAMP 含量的影响: 由表
2可见, Car 致炎后, 灌洗液 Neu 内 cAMP 含量明
显降低( P< 0. 01) ; PnS 能剂量依赖性地增加 Neu
内 cAMP 含量, 且其大剂量组能使 Neu 内 cAMP
含量恢复到正常水平。Dex 也具有类似作用。
3　讨论
自由基是广泛存在于生物体组织细胞内的非特
异性损伤因素,也是炎症发生发展的重要病理机制
之一[ 2]。炎症时,活化的白细胞呼吸暴发过程产生的
O
÷
2等活性氧自由基,不仅具有增加血管通透性的作
用,而且极易与生物膜的多不饱和脂肪酸发生脱氢
反应, 从而诱发组织细胞的脂质过氧化损伤[ 10]。本
实验结果发现, PnS 3 个剂量组均能剂量依赖性地
抑制 Car 致炎大鼠 Neu 释放 O÷2 、降低灌洗液中脂
质过氧化物 MDA 的含量,且与其降低 Neu 数量及
蛋白含量呈平行性变化。这与 Pns 对体外多形核白
细胞呼吸暴发时产生的氧自由基及黄嘌吟氧化酶体
系产生的 O÷2 均具有清除作用的结果相一致 [ 11]。
细胞内的第二信使分子 cAMP 在调控炎症发
生发展中具有重要作用,胞内升高的 cAMP 可抑制
炎细胞释放自由基而产生抗炎活性[ 3]。本实验观察
到, Car 致炎后,气囊灌洗液中 Neu 内 cAMP 含量
明显下降 ( P< 0. 01) , PnS 3 个剂量组均能剂量依
赖性升高 Neu 内 cAMP 含量,相关分析结果显示,
PnS 升高 Neu 内 cAMP 含量与其抑制 Neu 释放
O
÷
2 呈显著负相关 ( r= - 0. 938 4, P< 0. 01)。上述
结果表明,升高 Neu 内 cAMP 从而抑制 Neu 释放
O
÷
2 、减轻脂质过氧化损伤是 PnS 发挥抗炎作用的
重要分子机制之一。
参 考 文 献
1　张晓丹,刘建军,马英丽,等 . 中医药信息, 1993, ( 5) : 28
2　M aeda H, Akaike T . Biochem ist ry Mosc, 1998, 63( 7) : 854
3　T akei K, Toku yama K, Kato M , et al. Pharmacology, 1998, 57
( 1) : 1
4　Edw ards J C W, Sedgw ick A D, W illoug hby D A, et al . J
Pathology, 1981, 134( 2) : 147
5　福州部队总医院编 . 临床医学检验 . 上海:上海科技出版社,
1978: 15
6　Low ry O H, Rosebrough N J, Farr A L, et al. J Biol Chem,
1951, 193: 265
7　向　荣,王鼎年 . 生物化学与生物物理进展, 1990, 17: 241
8　Braquet M , Lavaud P, Dormont D, et al . Prostaglandines,
1985, 29( 5) : 747
9　陈海鑫,李振甲 . 生物化学与生物物理进展, 1986, 5: 78
10　Ignatow icz E. Pol J Pharmacol, 1994, 46( 3) : 103
11　周金黄 . 免疫药理学进展—基础与临床 . 北京:中国科学技术
出版社, 1993: 170
( 2000-03-08收稿)
Studies on the Basic Principles for the Processing of Rhizoma Cibotii
Part Ⅰ Influence of Rhizoma Cibotii and Its Processed Samples on
Thrombin Induced Rabbit Platelet Aggregation
　　L iaoning College o f T CM ( Shenyang 110032)　　L i Jun
(李　军) , Jia T ianzhu (贾天柱) , Du Guangli and Han
Hongw ei
　　The Second Affiliated Hospital o f Dalian Medical Univer sity　　L iu Jinpeng (刘进鹏)
摘　要　比较研究了狗脊及其不同炮制品对凝血酶诱导的兔血小板聚集作用的影响, 各炮制品均有抑制血小板聚
集作用, 抗血小板聚集作用砂烫品> 盐制品> 酒蒸品> 单蒸品> 生品。
关键词　狗脊　凝血酶　血小板聚集作用
Abstract　　The influence of Cibotium barometz ( L . ) J . Sm . and its processed samples on thrombin-
induced platelet aggr egation in rabbits w as studied. T he results show ed that al l dif ferent ly pr ocessed
samples tested could inhibit platelet agg regat ion, w ith act ivit ies in the decreasing order of Rhiz oma Ciboti i
roasted in st irring sand> steamed af ter being salted> steamed after steeped in w ine> simply steamed> the
unprocessed crude Rhiz oma Ciboti i.
Key words　　 Rhiz oma Cibot ii　thrombin　platelet ag gregat ion
·678· 中草药　Chinese T raditional and Herbal Drug s　2000 年第 31卷第 9期
李　军　1993年 7月毕业于辽宁中医学院中药专业(英语班)后,留在辽宁中医学院中药系炮制教研室任教。1998年 7月获辽宁中医学院生药学专业硕士学位。曾参加研究了国家中医药管理局资助课题“中药狗脊炮制工艺及原理研究”。
　　 Rhiz oma Cibot ii ( RC) is the dried rhizome of
Cibot ium bar ometz ( L . ) J. Sm. It is a t radit ional
Chinese herbal drug in common use to inv ig orate
the liver and kidney , st reng then the bones and
muscles and reliev e rheumat ism , by driving aw ay
the pathogenic factors o f cold dampness and
mot ivate the jo int from its numbness and rigidity,
w hich is claimed to be specially good for the
elders. Since these therapeutic eff icacies may be
related to it s act ion by pr omot ing blo od circulat ion
and reso lving blood stasis, it w ould be just ifiable
to delve into the influence o f RC and its processed
samples on platelet agg regat ion.
1　Materials and Instrument
1. 1　Plant M aterials: The raw RC was bought
fr om Shenyang M adicinal M ater ials Company and
authent icated by Professo r Zheng Taikun at
L iaoning Co llege of T CM .
The pro cessed samples, including RC roasted
in st irring sand, steamed after being salted,
steamed af ter steeped in w ine, and simply steamed
w er e processed as described in the methods
recor ded by Wang Xiao tao
[ 1]
. They w er e all
gr ounded, including the crude RC, to pass 20-
meshe sieve for ex t ract ion and preparat ion into
sample solut ions Ⅰ and Ⅱ.
1. 2 　 Reagents: T hrombin w as bought f rom
Beijing Biolog ical Pr oduct Inspect ion Inst itute.
Aspirin, the posit ive control, w as obtained from
the Aff iliated Hospital of L iaoning College of
TCM .
1. 3 　 A nimals: Rabbits w ere supplied by the
Experiment Animal Center of Liaoning Colleg e of
TCM .
1. 4　 Inst rument : Agg regometer, model BS-631,
w as purchased from Beijing Biopharmaceut ical
Factory.
2　Method and Result
2. 1　Preparat ion of Sample Solut ions: 10 g of the
powdered crude and processed RC were w eighed
accurately and sparately ex t racted w ith 95%
alcoho l by r ef lux. Each individual alcoholic ext ract
w as filtered af ter cooling and the solvent recovered
as completely as po ssible to give a residue which
w as t reated w ith 4 times its amount of w ater and
left overnight . They were filt rated and
concentrated on the nex t day and the r esultant
residue w as disso lved in distilled w ater in a 5 mL
volumetr ic f lask to give sample so lut ions with f inal
concentrat ions equivalent to 10 g of crude or
processed RC per 5 mL. This w as designated as
solut ion Ⅰ.
Another 10 g port ions of crude or pr ocessed
RC were likew ise ext racted w ith 95% alcohol, t re-
ated similarly as above, but the f inal w ater
solut ions w ere fur ther subjected to ext ract ion w ith
ethyl acetate to remove the presence of any lipo-
phil ic contents. T he w ater lay er w as concentrated,
t ransferred to 5 mL vo lumetric flasks w ith dist illed
w ater to g ive so lut ions also w ith f inal
concentrat ions equivalent to 10 g of crude or
processed RC per 5 mL, w hich w er e designated as
solut ions Ⅱ.
2. 2　Preparat ion of Platelet Rich Plasma ( PRP)
and Platelet Poo r Plasma ( PPP) : Rabbits of either
sex ( n = 8, w eighing 2～3 kg) w ere anaesthet ized
by int raperitoneal ( ip) inject ion of urethane. T he
citr ated car ot id blood w as centr ifuged for 5 min at
1 000 r/ min to obtain PRP w ith platelet number s
about 2. 0 × 106/ mL, the rest blo od w as
cont inual ly centrifuged fo r 10 min at 3 000 r/ m in
to obtain PPP. 3. 8 percent of sodium cit rate
solut ion w as used as ant icoagulant .
2. 3　Mesurements and Results [ 2, 3] : Turbidimetr y
w as used. 0. 5 mL of PRP w as placed into each
turbidimetr ic tube. 25, 50
L of dist illed w ater
w ere added respect ively into the blank contr ol
tubes. 15
L of aspirin solut ion ( 3 g dissolved in
100 mL of normal saline solution) w as added into
the posit ive control tube. Solut ion Ⅰ and solution
Ⅱ wer e added into the test drug tubes. A ll tubes
w ere placed in the agg regometer and incubated for
2 min at 37 ℃. The thrombin ( 1. 70 mg / 100 units
dissolved in 3 mL o f no rmal saline solut ion) w as
added as platelet agg regat ion inducer to each tube.
T he cur ves of absorbance by thrombin-induced
platelet ag gregat ion w ere reco rded.
T he inhibitory percentage of platelet
ag gregat ion was calculated by the follow ing
formula:
　　I= A B- A S
AB
Where : I= Platelet agg regation inhibition ( % )
·679·中草药　Chinese T raditional and Herbal Drug s　2000 年第 31卷第 9期
A B = Curve o f absorbance ( mm ) o f blank
contr o l
A S= Curve of absorbance ( mm) o f sample
T he data w er e expressed as x ± s and analy zed
w ith analysis of v ariance. ( See T ab. 1)
Tab. 1　Inf luence of Rhizoma Cibotii and its Processed Samples on Thrombin Induced
Rabbit Platelet Aggregation ( n= 8, x±s)
Group Dosage　　
(
L )　　
S olut ion Ⅰ
Absorbance　　
( mm)　　
Inh it ition
( % )
S olut ion Ⅱ
Absorbance
( mm )
Inhibit ion Ⅱ
( % )
1 25 143. 4±11. 8
50 144. 1± 8. 9
2 15 65. 5±11. 5* * 54. 3
3 25 122. 2±22. 2 14. 8 131. 0±34. 8 8. 6
50 104. 4±16. 5* 27. 6 103. 3±36. 6* 28. 3
4 25 97. 8±11. 1* * 31. 8 90. 7±25. 7* * 36. 1
50 71. 8±19. 6* * 50. 2 60. 7±20. 4* * 57. 8
5 25 122. 2±33. 0 14. 8 102. 8±20. 5* * 28. 3
50 94. 7±27. 4* * 34. 3 82. 7±12. 9* * 42. 6
6 25 123. 6±22. 1 13. 8 116. 4±15. 8 18. 8
50 84. 6±18. 3* * 41. 3 77. 8±15. 2* * 46. 0
7 25 92. 8±19. 5* * 35. 3 109. 2±30. 6 23. 8
50 75. 4±19. 5* * 47. 7 68. 1±29. 6 52. 7
　　　　Statis t ical signif icance vs dist ill ed w ater: * P < 0. 01　* * P < 0. 001
　　　　1. dis t illed w ater　2. asiprin　3. crude RC　4. RC roas ted in st irring sand　5. RC sim ply s teamed　6. RC s teamed af ter steeped in
w ine　7. RC s teamed after being salted
　 　 Compared w ith t test , ther e is no
signif icantdif ference betw een the inhibition of
so lut ion Ⅰ and that o f solut ion Ⅱ.
3　Discussion
3. 1　Compared w ith the Blank-Contro l Group: RC
and all of it s pro cessed samples show ed significant
inhibitor y act ions on platelet agg regat ion, w ith
inhibitor y intensity of platelet agg regat ion in the
decr easing o rder of RC r oasted in st irring sand, RC
steamed after being salted, RC steamed af ter stee-
ped in w ine, RC simply steamed and the cr ude RC.
3. 2　The Inhibitory Act ivity o f So lut ion Ⅱ: the
w ater part af ter ethyl acetate ext raction, is slight ly
st ronger than that o f solution Ⅰ, no t subjected to
ethyl acetate ex t ract ion, though stat ist ically non
signif icant lead us to suppo se that the act ive
constituent for inhibition o f platelet agg regat ion is
w ater-soluble. Based on our previous study on the
“Basic Principles for the Processing o f RC”, a
research project g ranted by SPA C, w e have
detected the pr esence and est imated the contents of
pr otocatechuic acid and protocatechualdehyde in
various processed RC by HPLC ( rest ricted
publicat ion data ) , it w ould be justif iable to
speculate that the ant i-platelet act ivity may be
clo sely r elated w ith the presence o f these tw o
compounds, especially the decreasing order of their
contents w as found to be closely coinsident w ith
that o f the platelet inhibition act iv ity . Whether
such speculation can achieve a f irm stand, or there
are some other chemical const ituents responsible
fo r the diverse pharmacolog ical act iv it ies of RC,
need further study .
References
1　 Wang Xiaotao, Corpus ab out Method of Pr oces sing Chin ese
Materia Med ica of Pas t Ages ( modern part ) . (历代中药炮制法
汇典 . 现代部分 ) . Nanch ang : J iangx i S cient ifi c Techn ical
Publis her s, 1989: 105
2　Li Yik ui, et al . Ph armacolog ical Experimental Meth od ology of
Chinese Materia Medica ( 中药药理实验方法学 ) Shanghai:
Shangh ai Scient if ic Tech nical Publis hers. 1991: 94.
3　Sh an Chunw en, et al. Acta Ph armacologica Sin ica. 1998, 19( 2) :
1646
编辑部注:本文经我刊顾问、英文编审史玉俊研究员修改、审定。
( Reciev ed in Nov. 9th, 1999)
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·680· 中草药　Chinese T raditional and Herbal Drug s　2000 年第 31卷第 9期