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目的用过氧化氢(H_2O_2)诱导大鼠胰岛微血管内皮细胞(IMECs)凋亡,观察钙非依赖型磷脂酶A_2(iPLA_2)对其凋亡的影响。方法 siRNA技术抑制IMECs iPLA_2表达,DNA ladder法检测H_2O2诱导后的DNA片段,Annexin V-FITC/PI双染色检测细胞凋亡率,DIOC_6(3)染色检测细胞线粒体膜电位。结果 (1)siRNA-iPLA_2可明显抑制IMECs内iPLA_2 mRNA的表达。(2)H_2O_2诱导后细胞DNA片段化程度随诱导时间的增加而增多,抑制iPLA_2表达后细胞的梯状条带明显增亮。(3)干扰组和未干扰组、干扰阴性对照组相比凋亡率明显升高(P均<0.01)。(4)与未干扰组比较,干扰组细胞线粒体膜电位明显下降(P<0.1)。结论抑制iPLA_2表达可促进H_2O_2诱导的IMECs凋亡。
Aim To observe the effect of calcium-independent phospholipase A_2 (iPLA_2) on the apoptosis of rat pancreatic islet microvascular endothelial cells (IMECs) induced by hydrogen peroxide (H 2 O 2). Methods siRNA was used to inhibit the expression of iPLA_2 in IMECs. DNA ladder was used to detect the DNA fragments induced by H2O2. The apoptotic rate was detected by Annexin V-FITC / PI double staining. The mitochondrial membrane potential was measured by DIOC_6 (3) staining. Results (1) siRNA-iPLA_2 significantly inhibited iPLA_2 mRNA expression in IMECs. (2) The degree of DNA fragmentation induced by H 2 O 2 increased with the increase of induction time, and inhibited the expression of iPLA_2. (3) Compared with the control group, the apoptosis rate in the interference group and the non-interference group was significantly higher (P <0.01). (4) Compared with the non-interfering group, the mitochondrial membrane potential of the interfered group decreased significantly (P <0.1). Conclusion Inhibiting the expression of iPLA_2 can promote the apoptosis of IMECs induced by H_2O_2.