1,25(OH)2D3对高糖诱导的牛视网膜血管内皮细胞中表达水平变化及对细胞凋亡水平的影响VEGF

Influence of 1,25(OH)2D3 on expression of VEGF and apoptosis rate in bovine retinal capillary endothelial cells induced by high glucose

目的观察1,25(OH)2D3对高糖诱导牛视网膜血管内皮细胞(BRECs)中血管内皮生长因子(VEGF)的表达水平变化及对细胞凋亡水平的影响。方法将分离培养的BRECs分为三组,分别为正常糖组、高糖组和高糖处理组。正常对照组细胞培养液含5 mmol/L葡萄糖,高糖组细胞培养液含30 mmol/L葡萄糖,高糖处理组细胞培养液含30 mmol/L葡萄糖和50n M,1,25(OH)2D3。培养48 h后收集细胞蛋白。蛋白免疫印迹法检测细胞VEGF及细胞凋亡相关蛋白Bax和Bcl-2表达水平;PI/Hoechst双染色法检测细胞凋亡。结果相比于正常糖组,高糖组中VEFG水平和Bax/Bcl-2比值显著增加;而在高糖处理组中表达水平远远低于高糖组,差异均有统计学意义(P<0.05)。高糖的细胞凋亡水平高于正常糖组,而经过1,25(OH)2D3处理后,其细胞凋亡水平则有所下降,差异均有统计学意义(P<0.05)。结论 1,25(OH)2D3可以抑制高糖诱导BRECs中VEGF表达增加及细胞凋亡。

Objetive To investigate the Influence of 1,25（OH）2D3 on VEGF expression and cellar apoptotic level induced by high glucose in bovine retinal capillary endothelial cells（BRECs）. Methods The BRECs were treated by different culture medium with various glucose concentrations（5mmol/L glucose,30 mmol/L glucose, 30 mmol/L glucose＋50n M 1,25（OH）2D3）as normal glucose group, high glucose group and teatment group respectively.After the cells incubated for 48 hours,the protein expression of VEGF、Bax 、Bcl-2 was detected by Western blot; propidium iodide/Hoechst staining was used for Analysis of apoptosis.Results Compared with normal glucose group,the protein level of VEGF、Bax /Bcl-2 markedly increased. The upregulation of VEGF and Bax /Bcl-2 induced by high glucose was prevented by vitamin D3 treatment. The same tendency existed in apoptosis assay。 Conclusions1,25（OH）2D3 decreases high glucose-induced upregulation of VEGF and Bax /Bcl-2 in BRECs.