高糖对大鼠心肌微血管内皮细胞凋亡和迁移的影响及机制

Effects of High Glucose on Apoptosis and Migration of Cardiac Microvascular Endothelia Cells in Rats and Its Mechanism

目的 观察高糖对体外培养的心肌微血管内皮细胞(CMEC)的凋亡及迁移能力的影响及可能的分子机制。方法 将原代分离、培养的大鼠CMEC随机分为对照组、高糖组、抑制剂组(高糖+XMD8-92)。流式细胞计数检测细胞凋亡,划痕实验测定细胞迁移能力;蛋白免疫印迹实验测定细胞外信号调节激酶5(extracellular signal-regulated kinase5, ERK5)水平及ERK5磷酸化水平。酶联免疫吸附试验测定血管内皮生长因子(vascular endothelial growth factor, VEGF)水平。结果 与对照组比较,高糖组细胞凋亡率增加,迁移能力及VEGF表达水平明显降低,P-ERK5水平明显升高( P <0.05)。抑制剂组与对照组比较差异无统计学意义( P >0.05)。结论 高糖可促进CMEC凋亡,抑制内皮细胞迁移能力,其机制可能与高糖促进ERK5的磷酸化从而抑制VEGF的表达有关。

Objective To investigate effects of high glucose on apoptosis and migration of cardiac microvascular endothelia cells (CMEC) in vitro culture and its potential mechanisms. Methods CMEC, which were separated and cultured from primary rats, were randomly divided into control group, high glucose group and inhibitor group (high glucose+XMD8-92). Apoptosis was detected by flow cytometry, and cell migration ability was detected by scratch test. Westernblot was used to detect extracellular signal-regulated kinase5 (ERK5) level and phosphorylation ERK5 (P-ERK5) level. Enzyme linked immunosorbent assay was used to detect vascular endothelial growth factor (VEGF) expression. Results Compared with those in control group, in high glucose group, apoptosis rate was increased, and cell migration ability and VEGF expression were significantly decreased, while P-ERK5 level was significantly increased ( P <0.05). There were no significant differences between inhibitor and control groups ( P >0.05). Conclusion High glucose may promote CMEC apoptosis and inhibit VEGF migration ability, and its mechanism may be related to inhibition of phosphorylation ERK5 so as to inhibit VEGF expression.