高糖对人角膜上皮细胞创伤修复中Cyclin D1蛋白表达的影响

目的:研究高糖环境对人角膜上皮细胞损伤修复的影响,初步探讨Cyclin D1蛋白在高糖培养的角膜上皮细胞创伤愈合中表达的意义。方法:模拟糖尿病角膜病变的高糖微环境,人角膜上皮细胞经复苏、培养传代后,分别设置等体积蒸馏水的DMEM完全培养基的正常对照组和含25mmol/L葡萄糖的DMEM完全培养基高糖处理组的两组细胞,细胞长满后进行划伤刺激,倒置相差显微镜下观察比较各组细胞生长状况及其变化,于不同时间点(0、12、24、48和72h)利用Western blot检测分析高糖培养的角膜上皮细胞中Cyclin D1蛋白的表达,qRT-PCR检测Cyclin D1 mRNA水平相对表达情况。结果:体外高糖处理条件下,人角膜上皮细胞划伤后修复速度减慢,漂浮细胞增多,细胞重新贴壁少,细胞间距增加,随着高糖处理时间的延长,细胞状态变差,生长速度慢;正常组细胞损伤修复较快,细胞密度增大,且形态规则,细胞膜光滑。Western blot检测划伤刺激上调Cyclin D1的表达,但随着时间延长上调作用呈逐渐减弱,两组Cyclin D1最高表达量均出现在12h。高糖处理组Cyclin D1表达量低于正常对照组。qRT-PCR结果显示:高糖处理后,Cyclin D1 mRNA表达呈上调趋势,但随着高糖处理时间延长,上调作用减弱,且在48h处mRNA水平恢复至与对照组同一水平。结论:在角膜上皮细胞创伤愈合过程中,高糖呈负性调节,抑制Cyclin D1蛋白的表达,且与角膜上皮细胞增殖能力下降及凋亡相关。

Study on relationship between diabetic retinopathy and blood glucose, blood lipid

Objective: To provide scientific basis for the pathogenesis and prevention and control of diabetic retinopathy through studying on relationship between different stages of diabetic retinopathy and blood glucose, blood lipid andother related factors. Mehods: Collected 196 type 2 diabetes patients in a hospital branch ,it could be divided into NDR (n=180), NPDR (n=11) and PDR (n=5) and collected 40 control cases. Each object were surveyed fundus colorized photographyorfundus fluorescence angiography examination, physical and blood tests, and the results were statistically analyzed. Results: BMI, SBP, HbAlc, FPG, P2hPG, TG, HDL, DN and urinary albumin/creatinine ratio (UACR) between the four groups were statistically difference;SBP, HbAlc, P2hPG, TG and UACR were positively correlated relationship with different stage of DR;HDL was negatively related with different stage of DR. Conclusion: SBP, HbAlc, P2hPG, HDL and UACR were factors of different stages of DR, the SBP, HbAlc, P2hPG and TG were more higher and the HDL was more lower, the DR was more serious.