全 文 ：组胺刺激 30 min,与单纯应用组胺的比较 , K f和 Jv
值明显降低和 σ值的明显升高 ,差异明显 ( P <
0. 01)。说明当归补血汤对组胺造成的 EC单层对蛋
白质和大分子物质的通透性的增加具有较强的抑制
作用。
3　讨论
表 2　当归补血汤对含组胺的 5 g /L白蛋白 Hanks灌流液引起的 EC单层通透性增加的作用 (x+ s)
组　别 n Kf
(μL /min· cm2· kPa)
Jv
(μL /min· cm2· kPa) σ
对照组 5 4. 62± 0. 53 11. 16± 1. 11 0. 52± 0. 02
组胺组 7 6. 27± 0. 82* 14. 89± 2. 18* 0. 26± 0. 04* *
当归补血汤组 4 4. 53± 0. 78△△ 11. 38± 1. 81△ 0. 38± 0. 03* △
　　　　与对照组比较: * P < 0. 05　* * P < 0. 01;　与组胺组比较: △ P < 0. 05　△△ P < 0. 01
　　在内皮细胞膜上介导的炎症介质受体有组胺、
缓激肽、血小板激活因子 ( PAF)和细胞因子等。当这
些受体被激活后 ,引起内皮细胞的收缩 ,导致细胞旁
路的开放或跨细胞穿透的膜孔径增宽 ,随之使血管
内皮通透性增加 [7 ]。其通透性增加机制涉及到细胞
外 [Ca2+ ]的内流和细胞内 [Ca2+ ]的释放 ;信号转导
系统的激活 ,包括 G蛋白和磷脂酶 C( PLC)信号、
蛋白激酶 ( PKC)和酪氨酸激酶 ( TK)的激活、以及肌
球蛋白轻链 ( M LC)的磷酸化和去磷酸化等 [ 8、 9]。近
年来 , M LC的磷酸化和去磷酸化的研究倍受重视。
细胞外 [Ca2+ ]的内流和细胞内 [ Ca2+ ]的释放以及
MLC的去磷酸化能够诱导肌动-肌球蛋白间相互作
用 ,中心张力增加 ,引起细胞骨架的变化和细胞间裂
隙形成 [10 ]。应用 [Ca2+ ]内流阻断剂和外源性 cAMP
能够抑制炎症介质所致的内皮通透性增加 [9 ]。
本实验采用内皮单层加压灌注装置 ,精确地定
量 Starling方程中的各个参数 K f、 Jv 和 σ。 在此 ,
K f和 J v值反映的是内皮单层对液体和小分子物
质的通透性 ,σ值反映的是内皮单层对大分子物质
的通透性。内皮细胞单层经含组胺的 hanks液和含
组胺的 5 g /L白蛋白 hanks液灌流后 ,对液体和小
分子物质的通透性以及对蛋白质和大分子物质的通
透性均明显增高。内皮单层经组胺刺激后 ,应用当归
补血汤能够逆转因组胺引起的内皮单层的通秀性增
加 ,说明当归补血汤对组胺造成的 EC单层通透性
增加具有改善作用。根据本实验的结果 ,还不能确定
当归补血汤改善内皮单层的通透性的作用机制 ,进
一步的工作将着重研究当归补血汤在抗炎症作用中
的细胞和分子机制 ,特别是对信号转导系统的影响。
参 考 文 献
1　周件贵 .时珍国药研究 , 1996, 7( 4): 250
2　康　永 ,杜晓峰 ,许　强 . 中成药 , 1997, 19( 12): 38
3　张　勇 ,曹　英 . 实用中西医结合杂志 , 1995, 8( 9): 542
4　 Schw art z S M . In Vi tro, 1978, 14: 966
5　 Pos tlethw ai te A E, Syyd erman R, Kang A H. J Ex p M ed, 1976,
144: 1188
6　 Cooper J A, Peter J D V, Fred L M , et al . J Appl, Physiol,
1987, 62: 1076
7　Walter T, Zydi Z, Les zek G, et al . Pf lugers Arch, 1995, 430: 145
8　 Ikeda K, Utoguchi N, Makimoto H, et al . Inf lammation , 1999,
23: 87
9　 Van Nieuw Amerongen G P, Drai jer R, Vermeer M A, et a l.
Ci rc Res , 1998, 83: 1115
10　 Rabiet M J, Plantier J L, Rival Y, et al . Arterios ele Th romb
Vasc Biol , 1996, 16: 488
( 2000-01-31收稿 )
Studies on the Calcium Antagonist Action of Arctigenin
　　 Liaoning Colleg e o f TCM ( Shenyang 110032)　 Gao Yang , Kang Tingguo and Zhang Xiaoyu
Abstract　　 The calcium antagonist action o f arctigenin ( ACT) w as studied in o rder to v erify the ef-
fect of Fructus Arctii fo r th e reliev e of ex terio r syndrome. M uscular contraction of iso lated rat t rachea,
colon, pulmona ry a rtery and thoracic ao rta induced by KCl, that of guinea pig t rachea induced by CaCl2 ,
befo re and af ter the addition of ACT w ere assessed and thei r contraction-response curv es draw n and PD 2
calculated according to Scot t. The inhibi tion rate o f tw o-phase contraction o f guinea pig t rachea induced by
acety lcholine chloride ( Ach) in comparison wi th verapamil ( V ER) w as calculated. Results o f the study
show ed that ACT could non-compa titiv ely antagonize the muscular cont raction of the test specimens w ith
·758· 中草药　 Chinese Traditiona l and He rbal Drug s　 2000年第 31卷第 10期
高　阳　女 , 1975年出生 , 1998年毕业于辽宁中医学院中药系 ,现为该系 98级药理硕士研究生。
PD 2 of 4. 01, 5. 11, 5. 98 and 6. 05 respectiv ely. Similar to V ER, ACT could non-competi tiv ely antag onize
the isola ted guinea pig t rachea wi th PD 2 of 4. 04 and 5. 62 respectiv ely. Bo th of them could inhibi t the
fi rst phase contraction induced by Ach wi th inhibi tion rates of 66. 14% and 81. 42% respectively. It w as
concluded tha t ACT, as the activ e consti tuent o f Fructus Arct ii , relax ed smooth muscle contraction by
blocking the po tential dependant Ca
2+
channel and the internal release of Ca
2+
.
Key words　　 arctigenin　 smoo th muscle contraction　 calcium antagnist
摘　要　对牛蒡苷元 ( ACT )的钙拮抗作用进行探讨 ,为确认 ACT是牛蒡子解表功能的有效成分提供实验依据。描
记给 AC T前后 ,各标本对 KCl或 CaCl2诱发收缩的量效曲线 ,按 Sco tt法计算 PD 2;测定给 ACT后 ,标本对乙酰
胆硷 ( Ach)诱发的两相收缩的抑制百分率。 ACT对离体大鼠气管、结肠、肺动脉、胸主动脉平滑肌由 KCl引起的收
缩产生非竞争性拮抗作用 ,其 PD 2分别为 4. 01, 5. 11, 5. 98, 6. 05; ACT和维拉帕米 ( Ver apamil)相似 ,对离体豚鼠
气管平滑肌由 CaCl2引起的收缩产生非竞争性拮抗作用 ,其 PD 2分别为 4. 04, 5. 62;对 Ach诱发的两相收缩只明显
抑制第一时相收缩 ,抑制率分别为 66. 14% , 81. 42% . ACT对平滑肌的松弛作用可能是由于阻滞电压依赖性钙通
道和内钙释放所致 ; ACT是牛蒡子解表功能的有效成分。
关键词　牛蒡苷元　平滑肌收缩　钙拮抗剂
　　Fructus Arct ii , the dried f ruit of Arct ium lap-
pa L. (Compositae ) i s a t radi tional Chinese medi-
cine pungent in f lav o r but cooling in effect. It wa s
used as an antipyretic for the reliev e of syndrome
o f interna l heat. It acts to dispel ex terio r w ind
whi le clea r away internal heat; by promo ting the
dispersing function o f the lung fo r the complete
eruption o f the conjunctiva and ai r passage, thus
remov ing toxic ma terials and benefi ts the pharynx.
ACT, a lignan isolated f rom Fructus Arct ii ha s
been repo rted in li teratures to possess the thera-
peutic ef fect o f lowering elev ated blood pressure by
blocking the Ca
2+
channel
[1 ]
. Thus, i t is justifiable
to make a deeper inquiry into the Ca
2+
antag onist
ef fect of ACT.
1　Material and Method
1. 1　 Material: ACT w as isolated and purified
f rom Fructus Arctii to a puri ty o f 99. 8% by Dr.
Xu Zhaohui and Pro f. Yang Songsong o f Depar t-
ment of Phytochemist ry of this Col leg e. It w as dis-
solv ed in w ater to concentra tions of 8. 06× 10- 4 ,
2. 69× 10- 3 and 8. 06× 10- 3 mol /L wi th the addi-
tion of 60, 200 and 200 m l /L of abso lute alcohol
respectiv ely.
V ER, product of Shanghai Tianfeng Pharma-
ceutical Facto ry.
Ach, product of Shanghai Third Reagent Fac-
to ry.
Kreb s so lution; Ca
2+
f ree Kreb s so lution and
high po tassium Ca
2+
f ree Kreb s solution w ere pre-
pared af ter the modi fied Polster s phy siological salt
so lutions
[2 ] .
Wistar rat and guinea pig of ei ther sex w eigh-
ing ( 250± 50) g and ( 300± 50) g respectiv ely w ere
supplied by Liaoning Vivisection Center, Co lleg e
of TCM.
LW A-20 Fo rce displacement t ransducer,
product of Shanghai Medicine Elect ronic Inst ru-
ment Plant.
XW T-204 Potentiometric recorder, product of
Shanghai Dahua Inst rument and M eter Plant.
1. 2　 Method: The isola ted smooth muscle speci-
mens, prepared to thei r respectiv e sui table leng th
( trachea, f rom the low er end of larynx to i t s emi-
nence w ere sectioned into ring s of 4～ 5 mm seg-
ments; colon, fasted for 48 h, sectioned into 1 cm
segments; pulmonary ar tery and tho racic ao rta,
sectioned into 3～ 4 mm segments) , w ere suspend-
ed in a bath fi lled with 10 mL Kreb’ s solution
maintained at a temperature of ( 37± 1)℃ and aer-
ated w ith a mix ture of 95% O2 and 5% CO2 . The
specimens w ere connected to the force displace-
ment t ransducer and loaded wi th an ini tial tension
of 1 g. The solutions w ere renewed every 20 min
to maintain f resh ness. Af ter equi librium, the test
drug s w ere added cumula tively , the added amount
should not ex ceed 2% of the to tal v olume of the
nutrient bath. On each addi tion, the contraction of
·759·中草药　 Chinese Traditiona l and He rbal Drug s　 2000年第 31卷第 10期
the specimen w as reco rded wi th the potentiometric
recorder. The terminal pH was not significant ly
changed when the drug s w ere added. Same amount
o f alcohol used a s blank control showed no signi fi-
cant ef fect on the contraction o f the specimens.
Data w ere expressed in x± s and statistically com-
pared by t-test. PD 2 values w ere calcula ted ac-
cording to Sco t t. The max imal response of the con-
trol w as recorded as 100% , and the concentra tion-
response curv es ( CRC ) were const ructed acco rd-
ing ly.
2　 Results
2. 1　 Effects of ACT on the contraction of 4
smoo th muscle specimens induced by KCl: Con-
tractions induced by a final concentration of 30
mmol /L of KCl was used as the control to assess
the activi ties o f the specimens. Cumulativ e addi-
tion o f KCl to a final concentra tion( mmol /L ) o f 12
～ 44 for t rachea, 9. 0～ 24. 6 for co lon, 6. 0～ 39. 3
fo r pulmona ry artery and thoracic aorta were used
as auto-control fo r their respectiv e CRC. The final
concentra tions ( mol /L) of ACT added w ere 3. 226
× 10- 6 , 1. 075× 10- 5 and 3. 226× 10- 5 respective-
ly. They w ere incubated fo r 10 min and the addi-
tion of KCl w as repea ted as described above fo r
CRC da ta w hich were show n in Fig . 1～ 4. It could
be seen that ACT at the ultimate concentrations
mentioned above could shif t the KCl CRC to the
right , depicting i ts non-competitiv e Ca
2+
antag o-
nist ef fects. The calculated PD 2 were 4. 01, 5. 11,
5. 98 and 6. 05 respectiv ely.
2. 2　 Effects of ACT on CaCl2 induced CRC of
guinea pig trachea: ACT a t the ultimate concentra-
tions of 3. 226× 10- 6 , 1. 075× 10- 5 and 3. 226×
10
- 5
mol /L and V ER a t 5× 10- 7 , 1× 10- 6 and 2×
10
- 6
mo l /L also show ed non-competi tv e antag onist
ef fect ( Fig. 5 and 6) . The calcula ted PD 2 were
4. 04 and 5. 62 respectiv ely.
2. 3　 Effects of ACT on tw o-phase contraction in-
duced by Ach: ACT at concentrations of 1. 29×
10- 4 , 1. 72× 10- 4 mol /L and V ER at 7. 5× 10- 6 ,
and 1. 0× 10- 5 mol /L show ed inhibi tion of the
fi rst-phase contraction but no signi ficant effect on
second-phase contraction ( Fig. 7 ) . Thei r rates of
a-cont rol; b-3. 226× 10- 6 mol /L; c-1. 075× 10- 5 mol /L;
d-3. 226× 10- 5 mol /L
compared to the cont rol: * P < 0. 05　* * P < 0. 01
Fig. 1　 Ef fects of Arctigenin on the Contraction of Iso-
lated Rat Trachea Induced by KCl (n= 5, x± s )
a-cont rol; b-3. 226× 10- 6 mol /L; c-1. 075× 10- 5 mol /L;
d-3. 226× 10- 5 mol /L
compared to the cont rol: * P < 0. 05　* * P < 0. 01
Fig. 2　 Ef fects of Arctigenin on the Contraction of Iso-
lated Rat Colon Induced by KCl (n= 5, x± s )
inhibi tion w ere 49. 13% and 66. 14% by ACT, and
57. 46% and 81. 42% by V ER respectiv ely.
3　 Discussion
High K
+
caused smoo th muscle contraction by
opening the po tential dependent channels ( PDC )
on the cellula r membrane during depo larization to
promo te ex t racellular Ca
2+
influx
[3 ]
. The action of
ACT, simi lar to V ER is to block the PDC and in-
hibi t ex t racel lula r Ca
2+
inf lux. Thus, it suppresses
intracellular calcium mobi li zation rather than calci-
um entry via recepto r opera ted channels ( ROC)
[4 ]
.
Ca
2+
antagonists have been ex tensi tvely used
·760· 中草药　 Chinese Traditiona l and He rbal Drug s　 2000年第 31卷第 10期
a-cont rol; b-3. 226× 10- 6 mol /L; c-1. 075× 10- 5 mol /L;
d-3. 226× 10- 5 mol /L
compared to th e cont rol: * P < 0. 05　* * P < 0. 01
Fig. 3　 Ef fects of Arctigenin on the Contraction of
Isolated Rat Pulmonary Artery Induced by
KCl (n= 5, x± s)
a-cont rol; b-3. 226× 10- 6 mol /L; c-1. 075× 10- 5 mol /L;
d-3. 226× 10- 5 mol /L
compared to th e con trol: * P < 0. 05　* * P < 0. 01
Fig. 4　 Ef fects of Arctigenin on the Contraction of
Isolated Rat Thoracic Aorta Induced by
KCl (n= 5, x± s)
to t rea t cardiov ascula r diseases. Recently, i t wa s
reported tha t bronchial asthma was also related
w ith abnormal Ca
2+
movement. Ca
2+
antagonists
can relax bronchial smooth muscle and blo ck the
production of inf lamma to ry mediators o f the ai r
passage at dif ferent stages of asthma tic at tacks.
Mo reover, this kind of drug s may show a syner-
getic ef fect wi th o ther antiasthmatics and reduce
the dose of ho rmone used by ho rmone-dependent
a-cont rol; b-3. 226× 10- 6 mol /L; c-1. 075× 10- 5 mol /L;
d-3. 226× 10- 5 mol /L
compared to the cont rol: * P < 0. 05　* * P < 0. 01
Fig. 5　 Ef fects of Arctigenin on the Contraction of
Isolated Guinea Pig Trachea Induced by
CaCl2 (n= 5, x± s)
a-cont rol; b-5× 10- 7 mol /L; c-1× 10- 6 mol /L;
d-2× 10- 6 m ol /L
compared to the cont rol: * P < 0. 05　* * P < 0. 01
Fig. 6　 Ef fects of VER on the Contract ion of
Isolated Guinea Pig Trachea Induced by
CaCl2 (n= 5, x± s)
pa tients
[5 ] . As a new type o f Ca
2+
antagonist,
ACT meri ts to be developed as a drug of choice fo r
t rea tment of both ca rdiovascular diseases and
bronchial asthma.
ACT could relax thoracic ao rta, increase the
ci rcula ting blood volume of o rgans, such as skin
and relax th e smoo th muscle of pulmonary a rtery
and trachea. Numerous li tera tures and our ow n ex-
perience show ed that ACT could resist the at tack
·761·中草药　 Chinese Traditiona l and He rbal Drug s　 2000年第 31卷第 10期
a-ACT 1. 29× 10- 4 mol /L; b-ACT 1. 72× 10- 4; c-V ER 7. 5×
10- 6 mol /L; d-V ER 1. 0× 10- 5 mol /L
com pared to th e con trol: * P < 0. 05　* * P < 0. 01
Fig. 7　Ef fects of Arctigenin on the Intracellular Calci-
um-Dependent Contract ion ( n= 5, x± s )
o f some pathogens, such as bacteria , fungi and
vi rus due to i ts action to increase immunological
functions. In t radi tional Chinese medical concept ,
pulmonary functions and intestinal symptoms rep-
resent the ex terio r and interior manifesta tions of
diseases. The relax ation o f colon may facilitated
bow el movement to reliev e gast rointestinal co ld.
Resul t o f the present study show ed that ACT, the
mainactiv e principle of Fructus Arct ii , can reliev e
the ex terior syndrome wi th simul taneous improve-
ment of intestinal peristalsis. Determination of the
ACT content in Fructus Arct ii i s a sui table and fea-
sible quality cont rol cri terion to assess the quali ty
of the drug.
References
1　 Ichikaw a K, Kinoshi ta T, Nishibe S, et al . Ch em Pharm Bul l,
1986, 34: 3514
2　 Chen S H, Yin Z S, Ma B B, et al . Acta Ph arm Sini, 1988, 9
( 6): 533
3　 Tenner T J, Eu r J Pharmacol, 1981, 73: 289
4　 Macvicar. BA, 1984, 226: 1345
5　 Liu K, Guan Y Y. Chin Pharmacol Bull , 1992, 8( 2): 91
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 April 30th, 2000)
钩藤中不同成分降压作用的差异△
上海中医药大学中药研究所 ( 200032)　　宋纯清 　樊　懿　黄伟晖　吴大正　胡之璧
摘　要　目的: 比较钩藤中提取的异钩藤碱、钩藤碱、钩藤总碱及非生物碱部分的降压作用。方法:大鼠麻醉后经颈
总动脉插管记录外周血压和经股静脉微量输注实验用药。 结果: 实验表明钩藤中 4种成分的降压强度为异钩藤碱
( 42. 0% ) > 钩藤碱 ( 32. 1% ) > 钩藤总碱 ( 21. 3% )> 钩藤非生物碱 ( 12. 4% )。 结论: 提示钩藤中主要降压成分为异
钩藤碱和钩藤碱。
关键词　钩藤　钩藤碱　异钩藤碱　钩藤总碱　钩藤非生物碱　外周血压
Different Hypotentive Effects of Various Active Constituents Isolated from
Uncar ia rhynchophylla
　　 Institute o f Chinese Ma teria Medica , Shanghai Univ e rsity o f TCM ( Shanghai 200032)　 Song Chunqing , Fan Yi, Huang
Weihui, Wu Dazheng and Hu Zhibi
Abstract　　 Hypo tensive ef fects of rhynchophy lline, isorhynchophylline, total alka loid and non-alka-
loid fraction isola ted f rom Uncaria rhynchophylla ( Miq. ) Jacks. w ere compared. Change in peripheral
blood pressure was reco rded by inserting a catheter into th e righ t common caro tid a rtery o f anaethetized
ra t, w hile each of the four hypo tensiv e constituents w as injected individually into the femoral v ein by a mi-
croinfusion pump. Results o f the findings show ed that the four consti tuents inU. rhynchophylla displayed
di fferent hypotensiv e potency in the o rder of isorhynchophylline [ lowering of mean a rterial pressure
( M AP) by 42. 0% ] > rhynchophy lline ( low ering o f MAP by 32. 1% ) > to tal alkaloid ( lowering o f MAP
by 21. 3% ) > non-alkaloid f raction ( low ering of MAP by 12. 4% ) . It w as concluded that iso rhyn-
chophylline and rhynchophylline w ere th e main h ypo tensiv e consti tuents in U. rhynchophy lla.
·762· 中草药　 Chinese Traditiona l and He rbal Drug s　 2000年第 31卷第 10期
Address: Song Chunqing, Inst itute of Chines e Materia M edica, Shanghai University of TCM, Shanghai宋纯清　男 , 1964年毕业于南京药学院 ,上海中医药大学教授 ,博士生导师。 专长中药化学 ,现任中药研究所中药化学室主任。 1986年和
1993年先后在日本东京大学和美国普渡 ( Purdue)大学多年访问合作研究 ,近年负责和承担国家和上海市研究课题 8项 ,主要从事中药和中药复方药效物质基础、质量标准及其作用机制研究 ,发表论文 40余篇。 参加抗癌新药羟基喜树碱的研究 ,获得上海市重大科技成果奖。△国家中医药管理局“九· 五”攻关基金 ( 969030203)