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目的:观察高糖对人脐静脉内皮细胞c-Jun表达的影响。方法:提取、分离人脐静脉内皮细胞,体外培养至第3代,分别用5.5,11.0,22.0,44.0 mmol/L葡萄糖培养液培养内皮细胞,并设22.0 mmol/L甘露醇培养液培养者为对照组。以22.0 mmol/L葡萄糖分别作用0,0.5,1.0,2.0 h,应用RT-PCR和免疫细胞化学方法检测c-Jun mRNA和蛋白表达水平。结果:高糖以浓度依赖方式上调人脐静脉内皮细胞c-Jun mRNA的表达,22.0 mmol/L葡萄糖达到最强刺激效应,与5.5 mmol/L葡萄糖组及对照组比较差异均有统计学意义(P<0.01)。22.0 mmol/L葡萄糖刺激0.5 h c-JunmRNA表达升高,刺激1.0 h达高峰,2.0 h开始下降。22.0 mmol/L葡萄糖刺激2 h后,人脐静脉内皮c-Jun蛋白表达增加。结论:高糖可上调人脐静脉内皮细胞c-Jun mRNA和蛋白表达。
Objective: To observe the effect of high glucose on c-Jun expression in human umbilical vein endothelial cells. Methods: Human umbilical vein endothelial cells were isolated and cultured in vitro. Endothelial cells were cultured in the third passage and cultured with 5.5, 11.0, 2.04 and 44.0 mmol / L glucose respectively. The medium containing 22.0 mmol / L mannitol Control group. The cells were treated with 22.0 mmol / L glucose for 0, 0.5, 1.0, 2.0 h respectively. The mRNA and protein levels of c-Jun were detected by RT-PCR and immunocytochemistry. Results: High glucose increased the expression of c-Jun mRNA in human umbilical vein endothelial cells in a concentration-dependent manner. The highest stimulation effect was achieved with 22.0 mmol / L glucose, which was significantly different from 5.5 mmol / L glucose group and control group P <0.01). After being treated with 22.0 mmol / L glucose for 0.5 h, the expression of c-Jun mRNA increased and peaked at 1.0 h, then decreased at 2.0 h. After 22.0 mmol / L glucose stimulation for 2 h, the expression of c-Jun protein in human umbilical vein endothelial cells increased. Conclusion: High glucose can up-regulate c-Jun mRNA and protein expression in human umbilical vein endothelial cells.