Angiopoietin-1对高糖培养内皮细胞连接相关蛋白的影响

发布时间：2018-08-11 11:10

【摘要】：目的：观察高糖培养人脐静脉内皮细胞连接相关蛋白(occludin、VE-cadherin和connexin40)的表达变化,探讨Ang-1对高糖环境下这些内皮细胞连接相关蛋白的影响。方法：将体外培养的人脐静脉内皮细胞随机分为正常组(5.5 mmol/L葡萄糖),甘露醇组(5.5 mmol/L葡萄糖+24.5 mmol/L甘露醇),高糖组(30 mmol/L葡萄糖),Ang-1干预组(30mmol/L葡萄糖+0.25mg/L Ang-1),用相应的培养基培养24h、48h后收集细胞,分别用实时定量聚合酶链反应(Real-Time PCR)检测各组脐静脉内皮细胞occludin、VE-cadherin和connexin 40 mRNA的表达,免疫荧光、免疫组化方法检测各组脐静脉内皮细胞occludin、VE-cadherin和connexin 40蛋白的表达。结果：(1)正常组和甘露醇组细胞均有occludi、VE-cadherin和connexin 40 mRNA的表达,但两组间无明显统计学差异(P0.05)；高糖组细胞occludin、VE-cadherin和connexin 40 mRNA的表达均较正常组下调(P0.01),并呈时间依赖性(24h vs 48h,P0.05)；Ang-1干预组细胞occludin、VE-cadherin和connexin 40 mRNA的表达虽低于正常组(P0.05),却较高糖组表达上调(P0.05),(2)免疫荧光和组化显示occludin、VE-cadherin和connexin 40主要表达于胞膜和胞浆,正常组和甘露醇组细胞均有表达,但两组间无明显统计学差异(P0.05)；高糖组细胞occludin、VE-cadherin和connexin 40蛋白表达均较正常组下调(P0.01),并呈时间依赖性(24h vs 48h,P0.05)；Ang-1干预组细胞occludin、VE-cadherin和connexin 40蛋白表达虽低于正常组(P0.05),却较高糖组表达上调(P0.05),(3)细胞occludin、VE-cadherin和connexin蛋白表达水平彼此呈正相关。结论：高糖培养内皮细胞连接相关蛋白occludin、VE-cadherin和connexin 40表达呈时间依赖性下调；外源性给予Ang-1,可上调occludin、VE-cadherin和connexin 40的表达,改善高糖对内皮细胞连接的损伤。
[Abstract]:Aim: to observe the expression of VE-cadherin and connexin40 in human umbilical vein endothelial cells cultured with high glucose, and to investigate the effect of Ang-1 on them. Methods: cultured human umbilical vein endothelial cells were randomly divided into normal group (5.5 mmol/L glucose), mannitol group (5.5 mmol/L glucose 24.5 mmol/L mannitol) and high glucose group (30 mmol/L glucose) 30mmol/L glucose 0.25mg/L Ang-1. The cells were collected after culture for 24 hours and 48 hours later. The expression of VE-cadherin and connexin 40 mRNA in umbilical vein endothelial cells were detected by real-time quantitative polymerase chain reaction (Real-Time PCR), and the expression of VE-cadherin and connexin 40 protein in endothelial cells of umbilical vein were detected by immunofluorescence and immunohistochemistry. Results: (1) there was no significant difference between the normal group and mannitol group in the expression of connexin 40 mRNA, but there was no significant difference between the two groups (P0.05), the expression of ocdinine VE-cadherin and connexin 40 mRNA in the high glucose group was lower than that in the normal group (P0.01) in a time dependent manner (24 h vs 48 h P 0.05). The expression of VE-cadherin and connexin 40 mRNA in the Ang-1 intervention group was lower than that in the normal group (P0.05), but it was higher than that in the high glucose group (P0.05), (2). The expression of VE-cadherin and connexin 40 were mainly expressed in the cell membrane and cytoplasm in the Ang-1 intervention group, and in the normal group and mannitol group, the expression of VE-cadherin and connexin 40 were found in the cells of the normal group and mannitol group. However, there was no significant difference between the two groups (P0.05), the expression of VE-cadherin and connexin 40 in high glucose group was lower than that in normal group (P0.01), and it was time-dependent (P0.05). The expression of VE-cadherin and connexin 40 in the Ang-1 intervention group was lower than that in the normal group (P0.05), but the expression levels of VE-cadherin and connexin protein were positively correlated with those in the high glucose group (P0.05), (3). Conclusion: the expression of VE-cadherin and connexin 40 in endothelial cells cultured with high glucose was down-regulated in a time-dependent manner, and the expression of VE-cadherin and connexin 40 was up-regulated by exogenous Ang-1.
【学位授予单位】：遵义医学院
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R363

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