Effect of high glucose on cultured retinal Müller cell in vitro

Background Retinal Muller cells can offer nutrient and maintain the normal structure of retina.Researches showed that the abnormality of Muiller cells leads to retinal vascular disease.To explore the effect of high glaucoma on retinal Muller cells is of a very important significance for the study on diabetic retinopathy (DR).Objective This study was to investigate the effects of different concentrations of glucose on retinal Muller cells in vitro.Methods Retinal tissue was isolated from 1 10-day-oM clean SD rat.Mtiller cells were cultured by explant culture method and passaged in DMEM containing 20％ fetal bovine serum.The third generation of cells were obtained and identified using glial fibrillary acidic protein (GFAP) staning.Then,5.5,30.0 and 40.0 mmol/L glucose were added into the culture medium for 4 days respectively.The proliferation (A570) of Muller cells was detected by MTT,and apoptosis rate of Muller cells was calculated by flow cytometer to evaluate the effects of 5.5,30.0 and 40.0 mmol/L glucose to cell vitality.Results Cultured and passaged cells grew well with the spindle shape.The positive reactive cells were ＞95％ for GFAP.The A570 value of Muller cells was 0.24±0.01,0.21±0.03 and 0.20±0.02 in 5.5,30.0 and 40.0 mmol/L glucose group respectively,showing a significant difference among the three groups(F=6.755,P＜0.05).Compared with 5.5 mmol/L glucose group,As70 values were significantly lower in 30.0and 40.0 mmol/L glucose group (q =0.645,0.486,P ＜ 0.05).Apoptosis rates of Miiller cells were (26.40 ±0.25)％,(30.19±0.16)％ and (36.23±0.19)％ in 5.5,30.0 and 40.0mmol/L glucose groups,with a significant difference among them (F =294.530,P＜0.05),and those in 30.0 and 40.0 mmol/L glucose groups were significantly reduced in comparison with 40.0 mmol/L glucose group (q =0.754,0.484,P ＜ 0.05).Conclusions High concentration of glucose inhibits the viability and promote the apoptosis of retinal Muller cells at a concentrationdependent manner. Key words: Retina; Muller cell; Glucose; Cell culture; Proliferation; Apoptosis