Expression of VEGF165 and VEGF165b in human retinal pigment epithelial cells in hypoxia and high glucose environment / 国际眼科杂志(Guoji Yanke Zazhi)

ABSTRACT

·AIM:Through the expression of VEGF165and VEGF165bin human retinal pigment epithelial cells in vitro in artificial simulated hypoxia and high glucose environment, to discuss their roles in the development of diabetic retinopathy and the relationship between each other. ·METHODS: After normal inoculation and cultivation of human retinal pigment epithelial cells (RPE) in vitro, the cells was divided into the normal group (5. 56mmol/L glucose,without CoCl2),the hypoxia group(5.56mmol/L glucose + 150μ mol/L CoCl2), the high glucose group (25mmol/L glucose, without CoCl2), the combination group (25mmol/L glucose + 150μ mol/L CoCl2),a total of four groups. The RNA of each group was extracted respectively in 12h,24h,36h,and 48h. We used the MTT colorimetry to detect cell vitality and growth trend; RT-PCR method to detect VEGF165and VEGF165brelative expression of mRNA of RPE cells in four different time points. ·RESULTS:Hypoxia and high sugar environment limited proliferation of RPE cell division and cell vitality. After comparing cells of the same group in different time points, in the normal group there was no statistically significant different expression over time (P>0.05); the expression in the hypoxia group, the high glucose group and the combination group increased over time, the difference was statistically significant(P<0. 05). At the same time,differences of the expression between groups was not statistical significant in 12h (P > 0. 05); the difference was statistically significant in 24h,36h,48h(P<0.05). ·CONCLUSION: Cultured RPE cells can express VEGF165b normal. Lack of oxygen and high glucose can induce the increase of VEGF165 mRNA, at the same time reduces the VEGF165bmRNA expression.