三种滇产鼠尾草复方注射液抗心肌缺血再灌注损伤的研究
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摘要
目的：与复方丹参注射液(丹参+三七)比较，研究三种滇产鼠尾草、滇丹参、甘西鼠尾和褐毛甘西鼠尾分别与三七合用对大鼠心肌缺血再灌注损伤的保护作用，探讨这三种滇产鼠尾草注射液在抗心肌缺血再灌注损伤方面替代丹参的可行性，探讨复方制剂的协同作用。方法：在体实验采用冠脉结扎松扎法建立大鼠心肌缺血再灌注模型(缺血30min，再灌注60min)，实验过程中以Ⅱ导联记录心电图变化，实验结束后取血清测定MDA、LDH、CK、SOD、GSH-Px，并测定心肌梗塞面积。离体实验采用Langerdoff灌流法建立心肌缺血再灌注模型，低灌(流量：0.4ml／min)15min，再灌注30min，再灌注后收集灌流液测定MDA、LDH、CK、SOD、GSH-Px。全程记录心肌收缩幅度、冠脉流量和心率。结果：1)这四种复方制剂均能显著降低在体和离体缺血再灌注后MDA、CK、LDH水平，升高SOD和GSH-Px，并有一定的剂量相关性。2)这四种复方制剂在在体模型均能加快心率，减轻ST段抬高程度，减少心肌梗塞面积；增加离体大鼠再灌注心脏的心肌收缩幅度、冠脉流量。3)与文献资料比较表明，各复方制剂在降低CK水平上强于单方制剂。4)三种滇产鼠尾草复方制剂高剂量(4g／kg)在降低在体生化指标MDA(p＜0.05，0.01)上强于复方丹参；除复方滇丹参外，各复方制剂在升高GSH-Px上优于复方丹参制剂(p＜0.05)；复方甘西鼠尾在高剂量时升高GSH-Px强于丹参(P＜0.01)。结论：丹参和三种滇产鼠尾草复方制剂均有较好的抗心肌缺血再灌注损伤作用，复方制剂优于其单方制剂，存在抗大鼠心肌缺血再
灌注损伤的协同作用，三种滇产鼠尾草在抗心肌缺血再灌注损伤方面可代用丹参。其机制
可能抗氧自由基损伤、钙拮抗等有关。
Background: Danshen (Salvia miltiorrhiza Bunge, SM), Diandanshen (S. yunnanesis C. H. Wright, SY), Ganxishuwei (S. przewalskii Maxim., SP), Hemaoganxishuwei (S. przewalskii Maxim. Var. mandarinorum stib.), and Sanqi [Panax notoginseng (Burk.) F.H.Chen, PN] are traditional drugs, which have been used to treat cardiovascular diseases for many years in China. Objective: In order to find out the cardio protection induced by three compounded injections made from SY+PN, SP+PN, SPM+PN respectively, during ischemia/reperfusion in rats, compared with injection made from SM+PN. And to confirm the synergism of the compounded injections, and find out whether the three drugs, SY, SP, and SPM can be used as SM against cardio ischemia/perfusion injury or not. Methods: Two ischemia/reperfusion models were employed. In one mordel, we ligated rats' artery of left anterior descending (LAD) for 30 min, then let it loose for 60 min, and we recorded the electrocardiogram (ECG) during experiment. We examined maleic dialdehyde (MDA
), lactate dehydrogenase (LDH), creatine kinase (CK), superoxide dismutase (SOD), and glutathione peroxidase (GSH-Px) in rats' serum after reperfusion. In another model, a heart-perfusion model in vitro, we perfused every rat's heart at low rate of flow with Kreb's solution for 15min at first, then followed normal perfusion for 30min, also examined MDA, LDH, CK, SOD, and GSH-Px in the flow of effluent after reperfusion. Contractile force, perfusion flow, and rhythm were recorded during experiment. Results: 1) All the four injections were able to decrease MDA, CK, LDH, and increase SOD, GSH-Px, in a way of dose-dependence, in vivo and in vitro. 2) All the four injections were able to quicken heart rhythm, mitigate ST segment, and decrease infarcted area in vivo. And were able to alleviate contractile force during ischemia, and enhance myocardio constractile force, increase coronary flow in the state of reperfusion. 3) It was indicated that all the compounded injections decreased CK in vivo more powerfully tha
n all the single historical ones respectively. 4) The three compounded injections of SY, SP, SPM decreased MDA at high dose (4g/kg) more powerfully than that of SM (p<0.05,0.01), in vivo; and except for SY, increased GSH-Px more powerfully than that of SM (p<0.05). In vitro, at high dose, the compounded injection of SP more powerfully increased GSH-Px than that of SM (p<0.01). Conclusion: All the three compounded injections (from SY+PN, SP+PN, and SPM+PN, respectively) employ cardio protection against ischemia/perfusion injury, and are similar to injection from SM+PN, and are more effective than the single ones respectively, which suggest that there is synergism of the compounded injections and that the three drugs can be replacer for SM to protect myocardium from ischemia/reperfusion injury. The mechanisms of which may be associated with blocking calcic channels, deactivating oxygen free radicles.
灌注损伤的协同作用，三种滇产鼠尾草在抗心肌缺血再灌注损伤方面可代用丹参。其机制
可能抗氧自由基损伤、钙拮抗等有关。
Background: Danshen (Salvia miltiorrhiza Bunge, SM), Diandanshen (S. yunnanesis C. H. Wright, SY), Ganxishuwei (S. przewalskii Maxim., SP), Hemaoganxishuwei (S. przewalskii Maxim. Var. mandarinorum stib.), and Sanqi [Panax notoginseng (Burk.) F.H.Chen, PN] are traditional drugs, which have been used to treat cardiovascular diseases for many years in China. Objective: In order to find out the cardio protection induced by three compounded injections made from SY+PN, SP+PN, SPM+PN respectively, during ischemia/reperfusion in rats, compared with injection made from SM+PN. And to confirm the synergism of the compounded injections, and find out whether the three drugs, SY, SP, and SPM can be used as SM against cardio ischemia/perfusion injury or not. Methods: Two ischemia/reperfusion models were employed. In one mordel, we ligated rats' artery of left anterior descending (LAD) for 30 min, then let it loose for 60 min, and we recorded the electrocardiogram (ECG) during experiment. We examined maleic dialdehyde (MDA
), lactate dehydrogenase (LDH), creatine kinase (CK), superoxide dismutase (SOD), and glutathione peroxidase (GSH-Px) in rats' serum after reperfusion. In another model, a heart-perfusion model in vitro, we perfused every rat's heart at low rate of flow with Kreb's solution for 15min at first, then followed normal perfusion for 30min, also examined MDA, LDH, CK, SOD, and GSH-Px in the flow of effluent after reperfusion. Contractile force, perfusion flow, and rhythm were recorded during experiment. Results: 1) All the four injections were able to decrease MDA, CK, LDH, and increase SOD, GSH-Px, in a way of dose-dependence, in vivo and in vitro. 2) All the four injections were able to quicken heart rhythm, mitigate ST segment, and decrease infarcted area in vivo. And were able to alleviate contractile force during ischemia, and enhance myocardio constractile force, increase coronary flow in the state of reperfusion. 3) It was indicated that all the compounded injections decreased CK in vivo more powerfully tha
n all the single historical ones respectively. 4) The three compounded injections of SY, SP, SPM decreased MDA at high dose (4g/kg) more powerfully than that of SM (p<0.05,0.01), in vivo; and except for SY, increased GSH-Px more powerfully than that of SM (p<0.05). In vitro, at high dose, the compounded injection of SP more powerfully increased GSH-Px than that of SM (p<0.01). Conclusion: All the three compounded injections (from SY+PN, SP+PN, and SPM+PN, respectively) employ cardio protection against ischemia/perfusion injury, and are similar to injection from SM+PN, and are more effective than the single ones respectively, which suggest that there is synergism of the compounded injections and that the three drugs can be replacer for SM to protect myocardium from ischemia/reperfusion injury. The mechanisms of which may be associated with blocking calcic channels, deactivating oxygen free radicles.
引文
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[2] 黄泰康．常用中药成分与药理手册．中国医药科技出版社，北京；1994：125
[3] 郑虎占，董泽宏，余靖等．中药现代研究与应用(第二卷)．学苑出版社，北京：1997；1099-1100
[4] 黄泰康．常用中药成分与药理手册．中国医药科技出版社，北京：1994；127-134
[5] 中国科学院昆明植物所．云南种子植物名录(下册)．云南人民出版社，昆明；1984：1772-1775
[6] 何洪静，李惠兰，张荣平，等．滇丹参，甘西鼠尾，褐毛甘西鼠尾水提取液对鼠血栓形成及微循环的影响。天然产物研究与开发，2003：15(2)；144-151
[7] 闫彩珍．滇丹参，甘西鼠尾，褐毛甘西鼠尾药效学研究．昆明医学院硕士研究生学位论文，2001；6
[8] 闰彩珍，李惠兰，张荣平，等．滇丹参，甘西鼠尾，褐毛甘西鼠尾对离体豚鼠心脏的影响．昆明医学院学报，2002：(1)；14-17
[9] 秦剑．三种鼠尾草注射液对大鼠心肌缺血血流动力学的影响和对心肌缺血再灌注损伤的保护作用．昆明医学院硕士研究生学位论文，2003；6
[10] 胡建林，张荣平，李惠兰，等．3种滇产鼠尾草属植物丹参酮ⅡA的薄层扫描定量。中成药．2000；22(10)：723-724
[11] 中国医学科学院药用植物资源开发研究所，中国医学科学院药物研究所，等．中药志(第一册)．人民卫生出版社，北京；1982：
[12] 陈奇．中药药理研究方法．人民卫生出版社．北京；1993：522-525
[13] 金铭惠．病理生理学．人民卫生出版社．北京：1996；153-154
[14] 王志敏，陈立信，张月芳，等．丹参酮ⅡA磺酸钠对心肌和溶血的作用．生理学报．1980；32(1)：18-22
[15] 伍杰雄，陈俊秀．三七总皂甙的负性变频和变力作用．中国药理学报．1988；9(5)：
409-412
[16] 徐长庆，王孝铭，范劲松，等．丹参酮Ⅱ-A对豚鼠单个心室肌细胞跨膜电位及L-型钙电流的影响．中国病理生理杂志．1997；13(1)：43-47
[17] 朱甫祥，刘国美，杨志宏，等．氧反常心肌细胞内游离钙浓度的变化及丹参素和SOD的影响．中国病理生理杂志．1999；15(10)：877-879
[18] 熊志刚，陈俊秀，孙家均．三七总皂甙对心肌动作电位及慢钙通道电流的影响．中国药理学报．1989；10(2)：122-125
[19] 李洪泰，石琳．三七总皂甙对心肌细胞Ca2+内流的影响．中国药理学报．1990：11(3)：213-217
[20] Koksal C, Suzer O, Bozkurt A-K, et al. Comparison of enoximone, amrinone, or levosimendan enriched St. Thomas' hospital cardioplegic solutions used for myocardial preservation in isolated guinea pig hearts. Acta-Medica-(Hradec-Kralove). 2002; 45(3): 93-7
[21] Kusumoto K, Igata H, Abe A, et al. In vitro and in vivo pharmacology of a structurally novel Na+-H+ exchange inhibitor, T-162559. Br-J-Pharmacol. 2002; 135(8): 1995-2003
[22] 郑虎占，董泽宏，佘靖，等．中药现代化研究与应用(第二卷)．学苑出版社，北京：1997；1115
[23] 邓士贤，莫云强．云南三七总皂甙的降压作用及其机制．云南医药1987；(2)：116-119
[24] 张翼，周兆年．心肌细胞低氧损伤的钠、钾、钙离子机制．心功能杂志．1999；11(4)：946-249
[25] Maczewski M, Beresewicz A. Role of nitric oxide and free radicals in cardioprotection by blocking Na+/H+ and Na+/Ca2+ exchange in rat heart. Eur-J-Pharmaeol. 2003; 461(2-3): 139-47
[26] 苏晓华，王孝铭，焦选茂，等．大鼠心肌缺血再灌注线粒体内膜体电子质子偶联和氧化磷酸化变化及丹参素的作用．中国病理生理杂志．1996；12(2)：186-189
[27] Liu K, Abe T, Sekine S, et al. Experimental study on the scavenging effects of ginsenosides on oxygen free radicals using model of heterotopic heart transplantation in rats. Ann-Thorac-Cardiovasc-Surg. 1998 Aug; 4(4): 188-91
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