U0126对高糖作用下角膜上皮细胞活性的影响

Effect of U0126 on Corneal Epithelial Cell Activity Induced by High Glucose

【目的】 探讨U0126对高糖作用下人角膜上皮细胞(HCEC)活性的影响。【方法】 将HCEC分为正常组(5.5 mmol/L的葡萄糖),高糖组(50 mmol/L的葡萄糖)及U0126低、中、高浓度组(5,10,20 μmo/L的U0126+50 mmol/L的葡萄糖),并培养48 h。测定细胞活力、凋亡情况、ROS荧光强度、钙离子浓度、线粒体膜电位、天冬氨酸蛋白水解酶3(Caspase 3)及Caspase 9活性及Bcl-2、Bax、细胞色素C(Cyt C)蛋白表达。【结果】 与高糖组比较,U0126低、中、高浓度组细胞活力显著提高(P<0.01),细胞凋亡率、ROS荧光强度、钙离子浓度及Caspase3、9活性降低(P<0.01),线粒体膜电位升高(P<0.01),Bcl-2表达量上调(P<0.01),胞质中Cyt C表达量下调(P<0.01),U0126中、高浓度组Bax表达量下调(P<0.01)。【结论】 U0126可能通过线粒体途径抵抗高糖诱导的HCEC细胞凋亡。

【Objective】 To investigate the effect of U0126 on activity of human corneal epithelial cells (HCEC) induced by high glucose. 【Methods】 HCEC was divided into normal group (5.5 mmol/L glucose), high glucose group (50 mmol/L glucose) and U0126 low, medium and high concentration group (5, 10, 20 micron o/L, U0126 + 50 mmol/L glucose), and cultured for 48 hours. Cell viability, apoptosis, ROS fluorescence intensity, calcium concentration, mitochondrial membrane potential, aspartate proteolytic enzyme 3 (Caspase 3) and Caspase 9 activity, and the expression of Bcl-2, Bax and Cyt C protein were measured.【Results】 Compared with the high glucose group, the cell viability of U0126 low, medium and high concentration groups was significantly increased (P<0.01), the apoptotic rate, ROS fluorescence intensity, calcium concentration and Caspase 3, 9 activity were decreased (P<0.01), the mitochondrial membrane potential was increased (P<0.01), the expression of Bcl-2 was up-regulated (P<0.01), the expression of Cyt C in cytoplasm was down-regulated (P<0.01), and the expression of Bax in U0126 medium and high concentration groups was down-regulated (P<0.01).【Conclusion】 U0126 may resist apoptosis of HCEC cells induced by high glucose through mitochondrial pathway.