Effect of high glucose on cell viability of rat retinal Müller cells cultured in vitro and the mechanism

QU Hong; LI Quan

doi:10.3969/j.issn.1002-266X.2015.28.005

Abstract：Objective To observe the effect of high glucose on the cell viability of rat retinal Müller cells cultured in vitro and to investigate the mechanism.Methods Rats′retinal Müller cells of serial subcultivation were divided into N, G15 , G25 , and G35 groups which were cultured by DMEM medium or DMEM medium containing 15 mmol/L, 25 mmol/L and 35 mmol/L glucose for five days, respectively.MTT assay was applied to detect the optical density ( OD value which represents the cell viability) in all groups.The expression of hypoxia-inducible factor-1α( HIF-1α) and erythropoietin ( EPO) was detected by immunochemistry, immunofluorescence and enzyme linked immunosorbent assay.Results The OD values of the N, G15 , G25 and G35 groups were respectively 0.83 ±0.02, 0.55 ±0.02, 0.45 ±0.02 and 0.33 ±0.02, and significant difference was found among these four groups ( all P<0.01) .There were no positive cells of HIF-1αand EPO in the N group.Cells presented yellow positive expression of HIF-1αby immunochemistry and green fluorescence posi-tive expression of EPO by immunofluorescence in the G15 , G25 and G35 groups.The OD values of HIF-1αprotein were re-spectively 0.02 ±0.01, 0.58 ±0.02, 0.75 ±0.02 and 0.92 ±0.02 in the N, G15 , G25 and G35 groups, and significant difference was found among these four groups (all P<0.01).The OD values of EPO in the N, G15, G25 and G35 groups were respectively 0.03 ±0.01, 0.48 ±0.02, 0.33 ±0.02 and 0.25 ±0.02, and significant difference was found among these four groups (all P<0.01).Conclusion High glucose may decrease the cell viability of rat retinal Müller cells cul-tured in vitro and increase the expression of HIF-1αand EPO which may be one of its mechanisms.