High glucose enhances H_2O_2-induced apoptosis in bovine aortic endothelial cells  

高糖增强过氧化氢诱导牛主动脉内皮细胞凋亡(英文)

作　　者：郭迅[1] 刘文兰[1] 谌立伟[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1期43-47,共5页中国药理学报（英文版）

摘　　要：AIM: To investigate the effect of high glucose on hy-droperoxide ( H2O2 )-induced apoptosis in cultured bovine aortic endothelial cells (BAEC). METHODS: BAEC were cultured and passaged in normal glucose (5.5 mmol·L-1, NG) and high glucose (25 mmol·L-1, HG). Morphologic changes and quantification of apoptotic cells were determined under fluorescence microscope after H2O2- treated BAEC for 24 h with Hoechst 33258 staining. DNA fragmentation was visualized by agarose gel electrophoresis. The expression of phospho-p38 Ca2+-calmodulin dependent protein kinase (CCDPK, formerly called MAPK) was measured by Western blotting. RESULTS: H2O2 elicited typical apoptotic morphologic changes (chromatic condensation, nucleus fragmentation). At 100 -300 μmol·L-1, both NG- and HG-BAEC incubated with H2O2 for 24 h increased cell apoptosis and phos-pho-p38 CCDPK expression in a concentration-dependent manner. In HG-BAEC, H2O2 induced DNA fragmentation at a lower concentration than that in NG-BAEC, and the apoptotic cell count in HG-BAEC was also higher than that of NG-BAEC (P< 0.05). Similarly, the expression of phospho-p38 CCDPK induced by H2O2 was up-regulated in HG-BAEC ( P < 0.05). CONCLUSION: High glucose enhances H2O2-induced apoptosis in BAEC, which is related to high expression of phospho-p38 CCDPK.目的:研究高糖对过氧化氢(H_2O_2)诱导牛主动脉内皮细胞(BAEC)凋亡作用。方法:BAEC培养并传代于正常葡萄糖(5.5 mmol·L^(-1))和高糖(25mmol·L^(-1))中,经H_2O_2处理24 h后,Hoechst 33258染色,荧光显微镜观察形态学变化及凋亡细胞计数;琼脂糖凝胶电泳分析DNA降解,Western blot法检测磷酸化p38 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)。在100-300 μmol·L^(-1)范围内,正常糖和高糖BAEC经H_2O_2处理后,浓度依赖性诱导细胞凋亡和磷酸化p38 CCDPK表达。高糖条件下诱导BAEC DNA降解浓度低于正常糖BAEC,细胞凋亡率和磷酸化p38 CCDPK表达均显著高于正常糖组(p<0.05)。结论:高糖促进H_2O_2诱导BAEC凋亡,可能与其增强磷酸化p38 CCDPK的表达相关。

关 键 词：APOPTOSIS DNA fragmentation Ca2+ -calmudulin dependent protein kinase hydrogen peroxide GLUCOSE Western blotting vascular endothe-lium cultured cells 

分 类 号：R363[医药卫生—病理学]