酸敏感离子通道1a在视网膜色素上皮细胞氧化损伤过程中的作用  

Effect of Acid-sensing ion channel la in cultural human retinal pigment epithelial cells during oxidative stress

作　　者：谭健[1] 许益聘[1] 刘广鹏[1] 叶信海[1] 

机构地区：[1]上海市第十人民医院整形外科,200072

出　　处：《中华实验外科杂志》2012年第1期126-128,共3页Chinese Journal of Experimental Surgery

摘　　要：目的观察酸敏感离子通道1a（ASICla）在视网膜色素上皮细胞（RPE）氧化损伤过程中的作用。方法采用实时逆转录-聚合酶链反应（RT—PCR）以及Western blot的方法检测RPE在1mmol／LH2O2诱导的细胞氧化损伤过程中（0．5～4．0h），ASICla的表达变化；同时分别采用ASICs阻断剂amiloride和PCTxl，以及通过细胞转染ASICla，检测对该氧化损伤过程的影响，并应用噻唑蓝（MTT）比色法评价细胞的活性。结果实时RT—PCR以及Western blot的结果均显示RPE中ASICla在mRNA及蛋白水平均有表达。1mmoL／LH2O2处理RPE1、2、4h后，RPE中ASICla的蛋白表达明显降低至（70．5±11．0）％、（40．6±5．6）％和（45．6±7．6）％。PCTx1阻断ASIC1a后，RPE细胞存活率明显降低，至4h细胞存活率为（76．2±5．0）％（与对照组比较，P〈0．05），而过表达ASIC1a能够提高RPE氧化损伤过程中细胞的存活率，与H2O2处理组比较，4h组为（38．0±6．0）％比（16．2±2．0）％（P〈0．05）。结论ASIC1a在RPE中具有表达并对RPE的氧化损伤具有一定的细胞保护作用。Objective To investigated whether Acid-sensing ion channel l a （ASICla） is ex- pressed in the retinal pigment epithelium （RPE） cells and whether it is involved in the function of these cells. Methods Real time reverse transeription-polymerase chain reaction （RT-PCR） and Western blot-ting were used to analyze the ASICla expression in ARPE-19 cells during oxidative stress induced by hydrogen peroxide （H202 ）. Furthermore, blockage or overexpression of ASICla in RPE cells were obtained using inhibitors （ amiloride and PCTx1 ） or by the transfection of cDNA encoding hASICla. Cell viability was determined using the methyl thiazol tetrazolium （MTT） assay. Results The RT-PCR and Western blot results showed that both the mRNA and the protein of ASICla are expressed in RPE. Blockade of ASICs by amiloride in normal RPE cells resulted in cell death, indicating that ASICs play an important physiological role in RPE cells. Furthermore, overexpression of ASICla in RPE increased cell survival after oxidative stress induced by H2O2. Conclusion ASICla is functionally expressed in RPE cells and may play an important role in the physiological function of RPE cells in protecting them from oxidative stress.

关 键 词：酸敏感离子通道1a 视网膜色素上皮细胞 过氧化氢 

分 类 号：R77[医药卫生—眼科]