摘要
目的:研究乌头类有毒中药毒性成分乌头碱、新乌头碱和次乌头碱对心肌细胞膜受体作用的机制。
方法:本实验采用放射配基结合分析法(radioligand bind assay,RBA)研究乌头碱、新乌头碱和次乌头碱对心肌细胞肾上腺素能受体不同亚型的影响;用放射受体分析法(radioreceptor assay,RAA)研究乌头碱、新乌头碱和次乌头碱对心肌细胞IP3和cAMP/cGMP比值的影响,从而揭示乌头碱、新乌头碱及次乌头碱对心肌细胞肾上腺素能受体不同亚型及乌头碱对心肌细胞信号转导途径的影响。
结果:乌头碱对心肌细胞作用1分钟时,可引起心肌细胞a1—AR数量上调,但对a1—AR的亲和力无影响;对心肌细胞β1—AR、β2—AR的亲和力和受体数量均无影响。新乌头碱和次乌头碱对心肌细胞作用1分钟时,a1—AR、β1—AR、β2—AR的亲和力和受体数量均无影响。乌头碱、新乌头碱及次乌头碱在不同时间点作用于心肌细胞后,给药组IP3和cAMP/cGMP的比值与正常组相比均升高。
结论:
1.乌头碱对心肌细胞作用1分钟时,对a1—AR有影响,对β—AR无影响;新乌头碱及次乌头碱对心肌细胞作用1分钟时,a1-和β—AR均无变化。
2.乌头碱、新乌头碱及次乌头碱对心肌细胞内IP3和cAMP/cGMP均有影响。
3.乌头碱对心肌细胞的损伤可能是通过多信号转导途径实现的。
Objiective:To research the mechanisms of the venenons constituent of the Aconitum—aconitine、mesaconitine、hypaconitine on the membrane receptor of myocardial cell.
Methods:Researching the influence that aconitine mesaconitine and hypaconitine act on the adrenoceptor subtype of the myocardial cell by RBA. Researching the effect that aonitine masaconitine and hypaconitine act on the IP3 and cAMP/cGMP of the myocardial cell by RAA.On the basis of these experiment,it is revealed that effect that aconitine mesaconitine and hypaconitine act on the adrenoceptor subtype and signal transduction pathway of the myocardial cell.
Results:Aconitine acting on the myocardial cell produce up-regulation of a1-adrenergic receptors,but has no effect on the affinity of a1-adrenergic receptors. Aconitine has no effect on the affinity and quantity ofβ-adrenergic receptors of the myocardial cell.Mesaconitine and hypaconitine have no influence on the affinity and quantity of a1 andβ-adrenergic receptors.After aconitine mesaconitine and hypaconitien acting on the myocardial cell in different time,the IP3 and the ratio of the cAMP/cGMP of MAT increased than normal group.
Conclusion:
1.Aconitine has no effect on a1-adrenergic receptors,but no effect on the affinity and quantity ofβ-adrenergic receptors of the myocardial cell.Mesaconitine and hypaconitine have no influence on the affinity and quantity of a1 andβ-adrenergic receptors.
2.After aconitine mesaconitine and hypaconitien acting on the myocardial cell,the IP3 and the ratio of the cAMP/cGMP of MAT increased
3.The toxic effect of aconitine action on the myocardial cell may have multiple signal transduction pathway.
引文
[1]陈信义,李峨,候丽,贾枚,刘雪强.乌头类生物碱研究进展与应用前景评述[J].中国中医药信息杂志,2004,11(10):922
[2]黄泰康.常用中药成分与药理手册[M].北京:中国医药科技出版社,1994:921-928
[3]符华林.我国乌头属药用植物的研究概况[J].中药材,2004,27(2):149
[4]邓有娟,吕怀莺.从中药有效成分看特殊药传统煎法[J].浙江中西医结合杂志,2001,11(6):387
[5]杨晓敏.实用急性中毒手册[M].成都:四川科学技术出版社,1991,252
[6]李宗浩.现代急救医学[M].杭州:浙江科学出版社,1993,408
[7]孟琼华.乌头碱对心肌细胞毒性作用机制的研究.成都中医药大学博士学位论文.
[8]任引津,张寿林,倪为民,等.实用急性中毒全书[M].北京:人民卫生出版社,2003:487-857
[9]中华人民国和国卫生部药典委员会编.《中华人民国和国药典(一部)》[M].北京:化学工业出版社,2005,132-133
[10]中华人民国和国卫生部药典委员会编.《中华人民国和国药典(一部)》[M].北京:化学工业出版社,2005,26-27
[11]中华人民国和国卫生部药典委员会编.《中华人民国和国药典(一部)》[M].北京:化学工业出版社,2005,163-164
[12]李娅萍,田颂九,王国荣.乌头类药物的化学成分及分析方法概况[J].中国中医杂志,2001,26(10):659
[13]邹文铨,严锐,汪丹丹,等.附子生用与炮用的化学分析研究[J].中国中药杂志,1995,20(2):86-87
[14]国家中医药管理局《中华本草》编委会.中华本草精选本(上册)[M].上海:上海科学技术出版社,1996,476,484
[15]中国药科大学,中国医药科技出版社主编.中药辞海(第二卷)[M].北京:中国医药科技出版社,1996,467
[16]楼之岑,秦波主编.常用中药材品种整理和质量[M].北京:北京医科大学· 中国协和医科大学联合出版社,1995,144-145
[17]周远鹏.附子对心血管系统作用和毒性的比较[J].中西医结合杂志,1984,4(1):48
[18]Kosuge T,Yokora M.Cardioactive principle of Aconitum jap on icum Thunb[J].Chem Pharm Bull,1976,24:176
[19]Konno C,Shirasaki M,Hikinoh,et al.Cardioactive principle of Aconitum carmichaeli roots[J].Planta Med,1979,35:150-155
[20]Huang L Z,Zhang D Y,Wang C Y,et al.Synthesis of higenaming[J].Acta Pharm Sin(药理学报),1981,16(12):931-933
[21]黄能慧,周远鹏,刘文化,等.附子注射液和去甲乌药碱药理作用的比较[J].中国药理学报,1980,1:34
[22]张正,刘秀杰,陶忠华,等.去甲乌药碱对血液动力学的影响及耐受性与安全性[J].中华医学杂志,2002,82(5):352
[23]Kimura I,Makino M,Takamura Y,et al.Positive chronotropic and inotropic effects of higenamine and its enhancing action on the action-induced tachyarrhythmia in isolated Murine Atria Japan J Pharmacol,1994,66:75
[24]Hammond H K,Mokiman F D.Effects of dobutamine and arbutamine on regional myocardial function in porcine-model myocardial ische mia[J].J Am Coll Cardial,1994,23:475
[25]Chong W S,Lee Y J,et al.Comparison of inodilator effect oh hige menine,tetrahydroisoquinoline analogs,and dobutamine in the rat[J].Korean J Physiol Pharmacol,1998,2:323
[26]Zhou Y P.The pharmacologiceffects and toxicity of FU-ZI(Acontium carmichaeli Dex.)and its major constituents[Y].Acta Pharm Sin(药学学报),1983,18(3):394-397
[27]Chen D H,Liang X T.Studies on the constituents of blateral from aconitum carmichaeli Dex.(FUZI).Isolation and atructural determination of salsolinol[J].Acta Pharm Sin(药学学报),1982,17(10):792-974
[28]Han G Y,Liang H Q,Zhang W D,et al.Studies on the alkaloids and a new cardiac principle isolated from JIANGYOUFU-ZI(Aconitum carmichaeli Dex.) [J].Nat Prod Res Dec(天然产物研究与开发).1997,9(3):30-33
[29]徐礅海,赵洪峰,徐雅娟,等.四川江油生附子强心成分得研究[J].中草药,2004,35(9):964-966
[30]丁光霞,等.川附子水溶性提取物对“阳虚”小鼠免疫功能的影响[J].陕西中医,1995,(7):299
[31]Hikino Hiroshi,et al.附子的抗炎成分[J].国外医学中医中药分册,1980,3(2):52
[32]周远鹏,等.附子研究Ⅵ-附子中乌头碱及其有关化合物的药理作用[J].中药药理与临床,1992,8(5):145
[33]张贾沐.乌头碱和闹羊花毒素的镇痛作用以及并用东莨菪碱和阿托品后的增强现象[J].生理学报,1958,(2):98
[34]Hikino Hiroshi,et al.单胺类化合物对乌头碱镇痛作用的影响[J].国外医学中医中药分册,1981,3(3):56
[35]黄衍民,李成韶.乌头注射液对小鼠的镇痛作用及其药效动力学研究[J].中国药学杂志,2000,35(9):613-615
[36]王雅贤,贾宽,张德山,王桂山.乌头碱对小鼠免疫功能影响的实验研究[J].中医药信息,1989,(6):40-41
[37]汪沪双.乌头碱抗风湿作用的药效学研究概述[J].基层中药杂志,1996,10(3):45-46
[38]苏孝礼.乌头及其炮制品中粗多糖药理作用的研究[J].中药材,1991,14(5):27
[39]刘天培.中国乌头对血管的作用[J].药学学报,1966,13(4):250
[40]江苏新医学院编.中药大辞典(上册)[M].上海:上海人民出版社.1997
[41]区耀华.乌头碱对依赖钙调蛋白的环核苷酸磷酸二酯酶的拮抗作用[J].生物化学与生物物理进展,1988,5(4):290
[42]柯铭清编.中草药有效成分理化与药理特性[M].第二版.长沙:湖南科学技术出版社.1982:119
[43]曹小玉,刘川,彭成.90年以来附子研究的文献定量分析[J].成都中医药大学学报,2004,27(3):58-59
[44]谷润国,胡晓梅,王宏侠,黄艳红.附子中毒前后心电图及心室电位检测分析[J].福建医药杂志,2002,24(2):131
[45]瞿琼华,罗龙新.乌头类药物中毒46例心电图分析[J].实用医药杂志,2004,11(6):1029
[46]周佐林.再议乌头类药物中毒的原因及防范[J].时珍国药研究.1997,8(1):48
[47]陈永东.镁剂治疗乌头碱致恶性心律失常[J].医学文选,2000,19(1):65
[48]王浴生.中药药理与应用[M].北京:北京科技出版社,2000,592
[49]Mosmann T.Rapid colorimetrie assay for cellular growth and survival:application to prolife ration and cytotoxic assays[J].J Imuuno Methods,1983,65:55-63
[50]Graham RM,Perez LM,Hwa J,et al.α1-Adrenergie receptor subtypes Molecular structure,function and signaling[J].Cin Res,1996,78(5):737
[51]韩启德.α1肾上腺素受体3种亚型在心血管共存的生理与病理生理意义[J].北京医科大学学报,1999,31(2):97-102
[52]Fadida D,Shimoni Y,Giles WR.A novel effect of norepinephrine on cardiac cells is mediated by adrenoceptors[J].Am J Physiol,1989:256(Heart Circ Physiol 25):H1500-H1504
[53]Endoh M,Blinks JR.Actions of sympathomimetie amines on the Ca~(2+)transients and contrctions of rabbit myocardium:reciproenl changes in myofibrillar responsiveness to Ca~(2+) mediated through α- and β-adrenoceptors[J].Circ Res,1988,62(2):247-265
[54]I Zheng M,Han QD,Xiao RP.Distine β-adreneigic receptor subtype signaling in the heart and their pathophysiological relevance[J].Acta Physiologica Sinica,2004,56(1):1-15
[55]Akhther SA,Lutterell LM,Rlckman HA,et al.Targeting the receptor-Gq interface to inhibit in vim pressure overload myocartiial hypertrophy[J].Science,1998,280(5363):574-577[56]Rens-Domiano S,Harem HE.Structural and functional relation-shlps of heterotrimeric G-proteins[J].FASEB J,1995,9(11):1059-1066
[57]Dzimiri N.Regulation of β-adrenoceptor signaling in cardiac function and disease[J].Pharmacol Rev 1999,51(3):465-501
[58]Kaumann A J,Bartel S,Molcnaar P,et al.Action of β2-adrenergic receptors hastens relaxation and mediates phosphorylation of phospholamban,troponin Ⅰ and G-protein in ventricular myocardium from patients with terminal heart failure[J].Circulation,1999,9(2):65-72
[59]Xiao R P,Avdonin P,Zhou Y Y,et al.Coupling of beta2-ademoceptor to Gi protein and its physiological relevance in murine cardiac myocytes[J].Cire.Res,1999,84:43-52
[60]Xiao R P,Lakatta F G.Betal-adrenoceptor stimulation and beta2-adrenoceptor stimulation differ in their effects on contraction,cytosolic Ca~(2+),and Ca~(2+) currant in single rat ventricular cells[j].Cire.Res,1993,73:286-300
[61]Wehrens XH,Marks AR.Altered function and regulation of cardiac ryanodine receptors in cardiac disease[J].Trends Biochem Sic,2003,28:671-678
[62]Xiao R P,Avdonin P,Zhou YY,et al.Coupling of beta2-adrenoceptor to Gi proteins and its physiological relevance in routine,cardiac myocytes[J].Cire.Res,1999,84;43-52
[63]Xiao R P,Lakatta F.G.Beta1-adrenoceptor stimulation and beta2-adrenoceptor stimulation differ in their effects on contraction,cytosolic Ca~(2+),and Ca~(2+) current in single rat ventricular cells[J].Cire.Res,1993,73:286-300
[64]Mogamic H,Nakano K,Tepikin AV,at al.Ca~(2+) flow via tunnels in polarized cell:recharging of apical Ca~(2+) stores by focal Ca~(2+) entry through basal membrane patch[J].Cell,1997,88:49-54
[65]Luo Clt,Rudy Y,A dynamic model of the cardiac rentricular action potential:Ⅱafterdepolarizations triggered activity and potenitation[J].Circ Res,1994,74:1047-1113
[66]沈磊,吴玉林.钠钙交换体及其抑制剂在心律失常中的作用[J].中国临 床药理学与治疗学,2006,11(6):610-614
[67]郑广华.环核苷酸与临床[M].第一版,天津:天津科学技术出版社,1985,63
[1]李娅萍,田颂九,王国荣.乌头类药物的化学成分及分析方法概况[J].中国中药杂志,2001,26(10):659
[2]黄泰康.常用中药成分与药理手册[M].北京:中国医药科技出版 社,1994:921-928
[3]符华林.我国乌头属药用植物的研究概况[J].中药材,2004,27(2):149
[4]陈龙,马聘,蔡宝昌,陆跃明,吴皓.乌头碱对大鼠心肌细胞钙通道阻滞作用的单通道分析[J].药学学报,1995,30(3):168-171
[5]张世忠,吴博威.乌头碱与钾离子通道激动剂合用对离体大鼠心脏的正性肌力作用研究[J].中国药理学通报,2001,17(5):570～572
[6]潘群皖,汪萌芽.乌头碱抑制心脏收缩作用的时反应量-效关系分析[J].中药药理与临床,1997,13(1):21-23
[7]周远鹏.附子研究Ⅵ-附子中乌头碱及其有关化合物的药理作用[J].中药药理与临床,1992,8(5):145
[8]潘群皖,汪桐.乌头碱致心律失常毒性作用的剂量分析[J].皖南医学院学报,1996,15(3):198-200
[9]李石蓝.乌头附子的药理与毒理[J].陕西新医药,1979,8(3):54
[10]翟启辉译.乌头的药理作用[J].国外医学·中医中药分册,1988,10(5):26
[11]张贾沐.乌头碱和闹羊花毒素的镇痛作用以及并用东莨菪碱和阿托品后的增强现象[J].生理学报,1958,(2):98
[12]Hikino Hisoshi,et al.单胺类化合物对新乌头碱镇痛作用的影响[J].国外医学中医中药分册,1981,3(3):56
[13]郑平.乌头碱镇痛作用部位及其与中枢去甲肾上腺素能系统的关系[J].中国药理学报,1988,9(6):48
[14]Shu H,Arita H,Hayashida M,Sekiyama H,Hanaoka K.Effects of processed Aconiti tuber and its ingredient alkaloids on the development of antinociceptive tolerance to morphine[J].J Ethnopharmacol,2006 Feb 20;103(3):398-405
[15]江苏新医学院编.中药大辞典[M].上册.上海:上海科学技术出版社,1986,228.
[16]区耀华.乌头碱对依赖钙调蛋白的环核苷酸磷酸二酯酶的拮抗作用[J].生物化学与生物物理进展,1988,5(4):290
[17]段建红,杨瑞华,胡三觉.乌头碱对受损背根节神经元自发放电的作用[J].第四军医大学学报,2005,26(13):1153-1156
[18]张道启,陈婉梅.乌头碱对螯虾神经轴突膜电生理与物理特征及兴奋性的影响[J].济宁医学院学报,1992,15(3):18-22
[19]Kimura M,Muroi M,Kimura I,Sakai S,Kitaqawa I.Hypaconitine,the dominant constituent responsible for the neuromuscular blocking action of the Japanese-sino medicine "bushi"(aconite root)
[20]Sikino Hiroshi,et al.附子的抗炎成分[J].国外医学中医中药分册,1980,3(2):50
[21]高田洋.中乌头碱对角叉菜胶浮肿抑制作用的机理[J].国外医学中医中药分册,1980,3(2):50
[22]周京滋.附子、四逆汤镇痛、抗炎作用的药效动力学研究[J].中国中药杂志,1992,17(2):104
[23]宋东江.乌头碱类化合物抗炎作用研究进展[J].中草药,1990,21(11):43
[24]唐希灿.3—乙酰乌头碱的抗炎作用[J].中国药理学报,1984,5(2):85
[25]欧阳瑜.10种中草药抗炎有效成分对白三烯生物合成的影响[J].中药药理与临床,1991,7(1):24
[26]康乐,岳寿伟,王晓聆,方明.乌头碱直流电离子导入对大鼠关节炎的镇痛作用及其机制探讨[J].中华物理医学与康复杂志,2005,27(12):733-736
[27]蔡定芳,沈自尹,陈晓红,刘彦芳.乌头碱对大鼠下丘脑促肾上腺皮质激素释放激素含量的影响[J].中国中西医结合杂志.1996,16(9):544-546
[28]马健,樊巧玲,牧野充弘,木村正康.阳虚模型小鼠腹腔巨噬细胞Ia抗原表达及乌头碱的作用[J].中国中西医结合杂志,1995,15(9):544-546
[29]柯铭清编.中草药有效成分理化与药理特性[M].第二版,长沙:湖南科学技术出版社.1982,119
[30]王雅贤,贾宽,张德山,王桂山.乌头碱对小鼠免疫功能影响的实验研究[J].中医药信息,1989,(6):40-41
[31]马健,陆平成,牧野充弘,木村正康.乌头碱对小鼠腹腔巨噬细胞Ia抗原 表达影响的研究[J].中国药理学通报,1997,13(4):341-344
[32]汤新铭,孙桂芝.乌头碱抑瘤及抗转移的研究与治癌的观察[J].北京中医杂志,1986,(3):27
[33]黄永融.乌头抗癌研究概述[J].福建中医药,1991,22(1):54
[34]王华灵,韩培秀,徐世明.乌头碱对癌症疼痛的治疗效果[J].中国中西医结合杂志,1994,14(4):219
[35]李峨,陈信义.乌头碱抗耐药性人口腔鳞状上皮细胞作用的体外研究[J].中国中医药信息杂志,2004,11(2):103
[36]刘雪强,陈信义,王玉芝,唐勇.乌头碱对KBv_(200)细胞Pgp蛋白表达影响的研究[J].中医药学刊,2005,23(1):70-72
[37]陈永东.镁剂治疗乌头碱致恶性心律失常[J].医学文选,2000,19(1):65
[38]凌佩莹.乌头类药物中毒53例临床心电图分析[J].右江民族医学院学报,2000,22(6):957
[39]钟林,朱永福,黄云祥.乌头碱中毒30例临床救治[J].中国全科医学,2002,5(11):919
[40]张福才.大剂量阿托品抢救乌头碱中毒体会[J].中国乡村医生,1997,4(12):18
[41]李名亮.急性乌头碱中毒致严重室性心律失常21例[J].中国职业医学,2001,28(5):45
[42]姜伟玲.草乌中毒16例救治体会[J].现代实用医学,2002,14(12):652
[43]王芳,张丽娟,高顺梅.乌头碱类中毒的临床观察及护理体会[J].黑龙江医学,2002,26(2):139
[44]张丽,李晔.乌头碱中毒致精神障碍2例[J].包头医学院学报,2002,18(3):245
[45]赵初环,卢中秋,黄唯佳,等.血液净化治疗乌头碱中毒[J].中华内科杂志,2001,40(7):502
[46]徐红,林津,刘孟庄,沈桢巍.重度乌头碱中毒致血红蛋白尿2例[J].内科急危重症杂志,2002,8(3):128