高浓度葡萄糖体外对人晶状体上皮细胞整合素连接激酶表达的影响

目的:观察体外培养的人晶状体上皮细胞系SRA01/04在高浓度葡萄糖条件下细胞的增殖活性及整合素连接激酶(integrin-linked kinase,ILK)的表达变化。方法:用人晶状体上皮细胞系SRA01/04,在培养液中加入不同浓度的葡萄糖溶液,使糖浓度分别为5.5mmol/L(正常对照组),15.5mmol/L,30.5mmol/L和50.5mmol/L,并设立5.5mmol/L葡萄糖+25mmol/L甘露醇组(甘露醇对照组)。细胞经过处理0,6,12,24,48,72h后,用MTT法检测晶状体上皮细胞的增殖活性,并用Western blot法检测细胞在30.5mmol/L葡萄糖处理6,12,72h后ILK蛋白表达量的变化。结果:高糖作用下晶状体上皮细胞的增殖活力高于正常对照组和甘露醇组,在30.5mmol/L组最为明显,而且随高糖暴露时间的延长,在48,72h时细胞的增殖活力增加更显著。30.5mmol/L葡萄糖作用6,72h,晶状体上皮细胞ILK蛋白的表达明显增高,分别是正常组的1.25和1.28倍,而甘露醇并不引起ILK的表达增加。结论:高浓度的葡萄糖促进人晶状体上皮细胞的增生,使细胞中ILK的表达量增加,这些变化并不依赖于高糖引起的高渗透压改变。

Effects of high glucose on the expression of integrin-linked kinase in human lens epithelial cells

AIM:To investigate the effects of high glucose on the proliferation and the expression of integrin-linked kinase(ILK)in human lens epithelial cells in vitro.METHODS:Human lens epithelial cell line SRA01/04 was used.The cells were exposed to either 5.5(normal control),15.5,30.5,or 50.5mmol/L glucose or 25mmol/L mannitol plus 5.5mmol/L glucose(mannitol control)for 0,6,12,24,48 and 72 hours.The survival and proliferation of the cells were quantified by MTT assay.The ILK protein expression was analyzed with western blotting after exposure to 30.5mmol/L glucose for 6,12 and 72 hours.RESULTS:There was an increase in cell proliferation from 15.5 to 50.5mmol/L of glucose compared with the normal and mannitol controls.The peaks of cell prolifera-tion were in the group of 30.5mmol/L glucose,and at 48 or 72 hours exposure.ILK protein levels in SRA01/04 cells exposure to 30.5mmol/L glucose were 1.25 and 1.28 times higher than those in control cells at 6 hours and 72 hours exposure respectively.High concentration of mannitol did not upregulate the expression of ILK protein.CONCLUSION:High glucose may induce up-regulation of ILK and increase the cell proliferation in the human lens epithelial cells.These effects are not dependent on the osmotic stress resulting from elevating the concentration of glucose.