Effect Of Sep15 Gene Silencing On Glucose-Induced Integrin In Human Lens Epithelial Cells

Cataract is the main blinding disease. According to the reports, cataract makes the 43% human blind in the world, which seriously affects the life quality of human. However, as the improvement of living conditions, the incidence of diabetic cataract is higher and higher, which gets the attention of people. Normal proliferation, migration and differentiation of lens epithelial cell are the basis to ensure the normal development of the lens. Some evidence suggest that necessary trace element selenium plays an important protective role in lens and that, the endoplasmic reticulum selenoprotein Sep15 has an important role in maintaining lens transparency. To carry out the research in the pathogenesis and prevention of cataract has important scientific significance.Integrin alpha6 plays an important role in lens tissue morphology, proliferation and differentiation. In this paper, we choose the human lens epithelial cells(hLECs)SRA01/04 as the research object and use MTT, Western-Blot and RT-PCR method to discuss the effect of Sep15 gene silencing on glucose-induced Integrin in human lens epithelial cells. The main results are as follows:(1) We choosed MTT method to measure the effect of high glucose in h LECs. The results showed that when the glucose concentration was 45 mM, the cell damage was the most serious, which could obviously induce ER stress. However, when the glucose concentration was 25 mM, the cell viability was higher and the expression of Integrin alpha6 increased more significantly. It suggested that the optimal growth glucose concentration to h LE cells was 25 mM, which could as the optimal concentration. 40ng/mLbFGF could promote the differentiation of hLECs, but different time of bFGF treated cell could have different effect on Integrin mRNA and protein. RT-PCR and WB results showed that bFGF duration for 6h, Integrin mRNA expression were the highest and bFGF effecting time for 24 h, Integrin protein expression were the highest. When hLE cells were exposed for bFGF and glucose and the glucose concentration was 25 mM, the cell survival rate was the highest and the expression of Integrin alpha6 mRNA and protein were the highest. However as glucose concentration levels were elevated, the expression of Integrin was inhibited instead. The experimental results showed that high levels of glucose might seriously make damage to cells, which might lead to the occurrence of cataract.(2) We researched the effect of Sep15 gene silencing under high glucose on expression of Integrin in human lens epithelial cells. The results showed that Sep15 gene silencing aggravated the effect of Integrin induced by high glucose during the lens epithelial cells differentiation. The experimental results demonstrated that the Sep15 had an appropriate protection function for differentiation of lens epithelial cells, which could prevent the occurrence of cataract to some extent.