摘要
目的:探讨抗心肌肌球蛋白重链抗体(AMHCA)诱导心肌细胞凋亡的作用及其机制。方法:从急性心肌梗死患者血清中提取AMHCA,研究其对成年大鼠心肌细胞凋亡的影响。DNA末端标记膜联蛋白-V/PI复染色法观察和测量心肌细胞凋亡。分别用免疫印迹、膜片钳和激光共聚焦法检测凋亡相关蛋白P53和Bcl-2以及第二信使钙离子的表达和浓度。结果:AMHCA诱导的心肌细胞凋亡具有剂量依赖性。在此过程中,促凋亡的核蛋白P53促进心肌细胞凋亡,而抑凋亡的胞质蛋白Bcl-2抑制心肌细胞凋亡。同时,胞内钙离子浓度升高,而L型钙通道则没有影响。结论:急性心肌梗死患者体内的AMHCA可能是一种新的起动因子,可诱导心肌细胞凋亡。
AIM： To study the effect of human anti -cardiac myosin heavy chain antibodies（AMHCA） on rat cardiomyocyte apoptosis. METHODS： Rat cardiomyocytes were isolated by the method of enzymolysis. Apoptosis of the cardiomyocytes was observed and measured by DNA end labelling and Annexin - V/PI double - staining assay. The proteins levels of apoptosis related P53 and Bcl - 2 and the second messenger calcium were measured by Western blotting, patch clamp and confocal calcium imaging, respectively. RESULTS： AMHCA was able to induce cardiomyocyte apoptosis in a dose dependent manner. In the presence of AMHCA, apoptosis - accelerating nucleoprotein P53 promoted myocardial apoptosis, while apoptosis -inhibiting cytoplasmic protein Bcl- 2 inhibited myocardial apoptosis. Meanwhile, the concen- tration of cytoplasmic calcium was elevated. No effect of AMHCA on L - type calcium currents was observed. CONCLU- SION： As a novel triggering factor, AMHCA isolated from the patients with AMI induces cardiomyocyte apoptosis.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2011年第2期209-214,共6页
Chinese Journal of Pathophysiology
基金
国家973重点基础研究发展规划基金资助项目(No.2007CB512000
子课题No.2007CB512005)
