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作 者:陈雪梅[1] 张学东[1] 宗元娟[1] 沈强[1]
机构地区:[1]重庆医科大学附属第一医院眼科,眼科学重庆市市级重点实验室,重庆400016
出 处:《第三军医大学学报》2012年第1期70-73,共4页Journal of Third Military Medical University
基 金:重庆市卫生局重点项目(2011-1-029)~~
摘 要: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。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-oL 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-tx 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-ct 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-ot from cytoplasm to cytomembrane were significantly lower in PKC-cL inhibitor inhalation group than in 25.5 mmol/L glucose group ( P 〈 O. O1 ). Conclusion concentration can up-regulate the expression of TLR2 mRNA in ARPE-19 cells and promote the PKC-ot from cytoplasm to cytomembrane through the PKC-ct pathway High glucose translocation of
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