高糖对体外培养视网膜色素上皮细胞toll样受体2表达的影响.doc

高糖对视网膜色素上皮细胞TLR2表达的影响 陈雪梅 张学东 宗元娟 沈强 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 【摘要】目的 观察高糖对体外培养的ARPE-19细胞TLR2表达的影响及机制。方法各组ARPE-19细胞刺激48h,实验组（不同浓度D-葡萄糖糖：5.5mmol/L、 15.5mmol/L、25.5mmol/L)，PKC抑制剂组（25.5 mmol/L D-葡萄糖糖和10mmol/l Go6979 PKC-α药物抑制剂）,高渗对照组（5.5mmol/L D-葡萄糖和20mmol/L甘露醇）。qRT-PCR和Wesrern blot方法对比分析不同糖浓度组ARPE—19细胞TLR2蛋白和mRNA表达水平、PKC-α蛋白转位，以及PKC药物抑制剂对ARPE—19细胞在高糖环境下表达TLR2蛋白的影响。结果 与正糖组相比，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（P0.05），并且高糖组高于中度高糖组（P0.05）；PKC-α抑制剂可以消弱高糖上调表达ARPE-19细胞TLR2的作用（P0.05）。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。 【关键词】糖尿病视网膜病变；ARPE-19；Toll样受体2；高糖 中图分类号：R779.6 The expression of Toll-like receptor -2 in ARPE -19 by High glucose Chen Xuemei Zhang Xuedong Zong Yuanjuan shengqiang (Department of Ophthalmology,the First Hospotal,Chongqing400016,China; Medical University; Chongqing Key Laboratory of Ophthalmology) [Abstract] Objective To observe the effection of high glucose on expression of the Toll-like receptor -2 in ARPE-19 and its mechanism . Method ARPE-19 were divided into different glucose concentration grape (5.5 mmol/l、 15.5 mmol/l、25.5 mmol/l D-glucose) ， inhibition grape (10mmol/l Go6976, PKC-alpha pharmacological inhibitor, + 25.5 mmol/l D-glucose); and an osmotic control grape (20mmol/l mannitol+5.5 mmol/l D-glucose). ARPE-19 were cultured in different condition for 48h .Then Western blot and Real—Time Reverse Transcription—Polymerase Chain Reaction(RT—PCR)were respectively used to determine TLR-2 protein and mRNA 1evels、PKC-alpha translocation in different concentration glucose ,and effection of Go6976,PKC-alpha pharmacological inhibitor on expression of TLR-2 protien in high glucose. Results high glucose (15.5 mmol/l、 25.5 mmol/l) increased TLR2 mRNA and protein,and PKC-alpha translocation (P0.05),and the 25.5mmol/l groupe were higher than the 15.5mmol/l group(p0.05) .Pharmacological inhibition of PKC-alpha activity significantly decreased effection of high glucose on TLR2 protein in ARPE-19 (P0.05). C