Effects of p38 and p42/p44 CCDPK signaling on H_2O_2-mduced apoptosis in bovine aortic endothelial cells  

p38和p42/p44 Ca^(2+)-钙调蛋白依赖性蛋白激酶信号在过氧化氢诱导牛主动脉内皮细胞凋亡中的作用<英文>

作　　者：刘文兰[1] 郭迅[1] 郭兆贵[1] 

机构地区：[1]湖南医科大学分子药理研究室

出　　处：《Acta Pharmacologica Sinica》2000年第11期33-38,共6页中国药理学报（英文版）

摘　　要：AIM: To investigate the effects of p38 and p42/p44 Ca2+ -calmodulin dependent protein kinases (CCDPK) signaling on hydroperoxide (H2O2) -induced apoptosis in cultured bovine aortic endothelial cells (BAEC). METHODS: Morphologic changes and quantification of apoptotic cells were determined under fluorescence micro-scope after a 24-h treatment of BAEC by H2O2. Cell vi-ability was determined with MTT method. DNA frag-mentation was visualized by agarose gel electrophoresis. The expression of phospho-p38 and phospho-p42/p44 CCDPK was measured by Western blotting. RE-SULTS: H2O2 elicited typical apoptotic morphologic changes (chromatic condensation, nucleus fragmentation) and DNA fragmentation. At 100 - 500 μmol·L-1, in-cubation of BAEC with H2O2 for 24 h also induced phos-pho-p38 and phospho-p42/p44 CCDPK expression in a concentration-dependent manner. Interestingly, H2O2-induced apoptosis was markedly increased by preincuba-tion with U0126, a specific p42/p44 CCDPK inhibitor. However, SB203580, a specific p38 CCDPK inhibitor, enhanced the expresssion of phospho-p42/p44 CCDPK induced by H2O2, but had no effect on BAEC survival. CONCLUSION: p42/p44 CCDPK signaling appears to play protective roles in H2O2-induced apoptosis in BAEC, whereas p38 CCDPK is not the main signaling pathway mediating H2O2-induced cellular apoptosis.目的:研究p38和p42/p44 Ca^(2+).钙调蛋白依赖性蛋白激酶(CCDPK)信号通路对过氧化氢(H_2O_2)诱导牛主动脉内皮细胞(BAEC)凋亡的调节作用.方法:H_2O_2处理BAEC 24 h后,荧光显微镜下观察形态学变化及凋亡细胞计数.MTT法测定细胞活性,琼脂糖凝胶电泳分析DNA降解,蛋白质印迹法检测磷酸化p38和p42/p44 CCDPK表达.结果:H_2O_2诱导BAEC产生典型的凋亡细胞形态学变化(核浓染,核碎裂)和DNA断片.H_2O_2(100-500μmol·L^(-1))浓度依赖性刺激磷酸化p42/p44和p38 CCDPK的表达.p42/p44 CCDPK抑制剂U0126显著增强H_2O_2致凋亡作用;然而p38 CCDPK抑制剂SB203580可增强H_2O_2诱导的磷酸化p42/p44 CCDPK的表达,但不影响BAEC的存活.结论:p42/p44 CCDPK对H_2O_2诱导的BAEC凋亡起保护作用,而p38 CCDPK不是介导H_2O_2所致细胞凋亡的主要信号通路.

关 键 词：APOPTOSIS DNA fragmentation Ca2+-calmodulin dependent protein kinase hydrogen peroxide Western blotting vascular endothelium cultured cells 

分 类 号：R363[医药卫生—病理学]