山莨菪碱对心脏骤停猪内质网源性心肌细胞凋亡的保护

Protection of anisodamine on endoplasmic reticulum-related myocardial cell apoptosis in cardiac arrest swine

目的:通过观察心脏骤停家猪心肌氨基末端激酶(JNK)、caspase-12、钙蛋白酶(calpain)mRNA水平及蛋白表达,以及超微结构的变化,探讨山莨菪碱对心肌内质网源性调亡的保护机理。方法:用随机数字表法将23只健康家猪分为干预组(n=9),对照组(n=9),假手术组(n=5)。电致颤诱发心室纤颤,持续9min后进行标准心肺复苏,干预组为复苏过程中联合使用山莨菪碱。自主循环恢复后24h取心肌组织,透射电镜下观察心肌超微结构改变,RT-PCR检测JNK、caspase-12、calpain mRNA水平及蛋白含量变化。结果:与对照组比较,干预组心肌纤维排列较规则,心肌细胞内的内质网、线粒体和细胞核损伤轻,山莨菪碱减轻心肌细胞超微结构损伤。干预组Calpain mRNA水平较对照组减少[(0.25±0.04与(0.34±0.07),P<0.05];干预组JNK、caspase-12和calpain mRNA蛋白表达介于假手术组和对照组之间,各组间差异均有统计学意义。结论:山莨菪碱可能通过抑制内质网源性细胞凋亡的信号通路,对心脏骤停后缺血再灌注心肌有保护作用。

Objective：Investigate the protection of anisodamine on endoplasmic reticulum （ER）-related myocardial cell apoptosis in cardiac arrest swine, by observing the mRNA amount and protein expression of myocardial J NK, Caspase-12 and Calpain, and the change of cardiac myocyte ultrastructure. Method：By random digits table, 23 health Swines were divided into three groups, treatment group （n ~ 9）, control group （n~ 9） and sham group （n 5）. By alternating current stimulating, ventricular fibrillation was induced and last 9 minutes, then the standard car- diopulmonary resuscitation （CPR） was performed. In treatment group, anisodamine was combined used with epi- nephrine during Crafter resumption of spontaneous circulation （ROSC） for 24 h,the swine hearts were harvested. The myocardial ultrastructure were observed by transmission electron microscopy, and the mRNA amount and protein expression of myocardial JNK, Caspase-12 and Calpain were detected by reverse transcriptionpolymerase chain reaction （RT-PCR） and Western-Blot. Result： Contrast to the control group, the cardiac muscle fibers arranged more regular with light injured of ER,mitochondria and nucleus in myocardial cell, anisodamine could re- duce the cardiac myocyte ultrastructure damaged degree. The Calpain mRNA amount in treatment group was less than that in control group （0.25±0.04 vs 0. 34±0. 07,P〈0. 05）. The protein expression of myocardial JNK, Caspase-12 and Calpain in treatment group were higher than sham group, and lower than control group, there are statistic difference. Conclusion： By inhibiting ER-related myocardial cell apoptosis pathways, anisodamine can protect the myocardia from ischemia-reperfusion injured after cardiac arrest.