高糖对视网膜色素上皮细胞Toll样受体2表达的影响

目的观察高糖对体外培养的ARPE-19细胞Toll样受体2(toll-like receptor 2,TLR2)表达的影响及机制。方法各组ARPE-19(RPE细胞系)细胞分别刺激48 h,正常糖浓度组(5.5 mmol/L D-葡萄糖)、中度高糖组(15.5 mmol/LD-葡萄糖)、高糖组(25.5 mmol/L D-葡萄糖)、PKC-α抑制剂组(25.5 mmol/L D-葡萄糖加10 mmol/L Go6976 PKC-α抑制剂)、高渗对照组(5.5 mmol/L D-葡萄糖加20 mmol/L甘露醇)。Western blot检测各组ARPE-19细胞TLR2蛋白表达水平,以及PKC-α蛋白胞浆向胞膜的转位情况;Real-time PCR分析各组ARPE-19细胞TLR2的mRNA表达水平。结果与正常糖浓度组相比,高糖组和中度高糖组TLR2的mRNA和蛋白表达,以及PKC-α转位均增加(P<0.05),并且高糖组高于中度高糖组(P<0.05)。与高糖组比较,PKC-α抑制剂组的TLR2蛋白表达降低(P<0.05),PKC-α蛋白转位显著降低(P<0.01)。结论高糖可以上调ARPE-19细胞TLR2的mRNA和蛋白表达,刺激PKC-α转位;高糖可能是通过PKC-α通路上调ARPE-19细胞TLR2表达。

Effect of high glucose concentration on expression of toll-like receptor-2 in ARPE-19 cells

Objective To observe the effect of different glucose concentrations on expression of toll-like receptor-2(TLR-2) in ARPE-19 cells.Methods ARPE-19 cells were divided into 5.5 mmol/L glucose group,15.5 mmol/L glucose group,25.5 mmol/L glucose,25.5 mmol/L glucose +10 mmol/L Go6976 PKC-α inhibition group,5.5 mmol/L glucose + 20 mmol/L mannitol control group.Expression level of TLR-2 in ARPE-19 cells and translocation of PKC-α from cytoplasm to cytomembrane were measured by Western blotting,while that of TLR-2 mRNA in ARPE-19 cells was measured by real-time PCR.Results The expression level of TLR-2 mRNA and the translocation of PKC-α from cytoplasm to cytomembrane were higher in 25.5 mmol/L glucose group and 15.5 mmol/L glucose group than in control group(P<0.05).The expression level of TLR-2 protein and the translocation of PKC-α from cytoplasm to cytomembrane were significantly lower in PKC-α inhibitor inhalation group than in 25.5 mmol/L glucose group(P<0.01).Conclusion High glucose concentration can up-regulate the expression of TLR2 mRNA in ARPE-19 cells and promote the translocation of PKC-α from cytoplasm to cytomembrane through the PKC-α pathway.