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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。
AIM: To investigate the effect of high glucose on apoptosis of bovine aortic endothelial cells (BAEC) induced by hydrogen peroxide (H 2 O 2). Methods: BAECs were cultured and passaged in normal glucose (5.5 mmol·L -1) and high glucose (25 mmol·L -1) for 24 h. Hoechst 33258 staining and morphology were observed by fluorescence microscopy The changes of apoptotic cells and the number of apoptotic cells were analyzed. DNA degradation was analyzed by agarose gel electrophoresis. The expression of phosphorylated p38 CCDPK was detected by Western blot. Results: H 2 O 2 induced typical morphological changes of apoptotic cells (nuclear staining and nuclear fragmentation). In the range of 100-300 μmol·L -1, normal glucose and high glucose BAECs were treated with H 2 O 2 to induce apoptosis and phosphorylation of p38 CCDPK in a concentration-dependent manner. Under high glucose conditions, the BAEC DNA concentration was lower than that of normal BAEC, and the apoptotic rate and phosphorylated p38 CCDPK expression were significantly higher than those of normal glucose group (p <0.05). Conclusion: High glucose can promote the apoptosis of BAEC induced by H 2 O 2, which may be related to the enhancement of phosphorylated p38 CCDPK expression.