钙非依赖型磷脂酶A2对过氧化氢诱导人胰岛微血管内皮细胞凋亡的影响

Influence of calcium-independent phospholipase A2 on the H_2O_2-induced apoptosis of islet microvascular endothelial cells

目的研究钙非依赖型磷脂酶A2(iPLA2)对过氧化氢(H2O2)诱导的胰岛微血管内皮细胞(IMEC)凋亡的影响。方法实验分空白对照组、重组腺病毒空载体组(Ad-GFP组)、Ad-iPLA2组三组进行,每组均接受100μmol/L H2O2和500μmol/L H2O2两个干预浓度,作用时间12h,每组细胞做6个复孔(n=6);采用Hoechst33258核染色形态学观察、定量分析以及Annexin V-APC/P I双染色检测细胞的凋亡情况。结果空白对照组与空载体组比较,在H2O2浓度(100、500μmol/L)中细胞凋亡率差异均无统计学意义(P均>0.05);iPLA2组在100μmol/L H2O2浓度下凋亡率为(17.16±1.53)%,明显低于其他两组(P均<0.05);而在500μmol/LH2O2浓度下iPLA2组凋亡率(46.18±2.12)%,明显高于其他两组(P均<0.05)。结论 iPLA2在100μmol/L H2O2浓度下可抑制IMECs的凋亡,而在500μmol/L H2O2浓度下则促进IMECs的凋亡。

Objective To investigate the influence of calcium-independent phospholipase A2 （iPLA2） on the apoptosis in islet microvascular endothelial cells （IMEC） induced by hydrogen peroxide （H2 O2）. Methods Human IMECs were divided into 3 groups （n = 6）： the control group, Ad-GFP group, and Ad-iPLA2 group. They were treated by different concentrations of H202 （100 and 500 μmol/L） for 12 h. The apoptosis detection was performed by morphologic observation and quantitative analysis of IMECs stained with Hoechst33258 and by Annexin V APC/PI staining by FCM in each group. Results The comparison between the control group and Ad-GFP group showed that there was no significant difference in the apoptosis ratio （all P〉0.05）. The apoptosis ratio of Ad-iPLA2 group [（17.16 ±1.53）%] was lower than the other two groups （P〈0.05） in H2O2 100 μmol/L, but it was higher [（46.18±2. 12）%] than the other two groups （P〈0. 05） in H2O2 500 μmol/L. Conclusion The iPLA2 may inhibit the apoptosis of IMECs induced by H202 100 μmol/L, but may promote the apoptosis of IMECs induced by H2 02 500 μmol/L.