金诺芬通过EGFR/p38MAPK信号通路抑制视网膜色素上皮细胞活力  

Inhibition of auranofin on cell viability of retinal pigment epithelium by EGFR/p38MAPK signaling pathway

作　　者：陈晓冬 王彤[1] 晁媛媛 CHEN Xiao-dong;WANG Tong;CHAO Yuan-yuan(Shaanxi Ophthalmological Institute/Ophthalmology Department,Xi＇an No.1 Hospital,Xi＇an 710002,China)

机构地区：[1]陕西省眼科研究所/西安市第一医院眼科,陕西西安710002

出　　处：《临床医学研究与实践》2018年第32期1-4,共4页Clinical Research and Practice

基　　金：陕西省自然科学基础研究计划面上项目(No.2018JM7040)

摘　　要：目的探讨金诺芬(AF)抑制视网膜色素上皮(RPE)细胞活力的作用机制。方法体外培养人视网膜色素上皮细胞株(ARPE-19细胞),经2.0μM AF刺激ARPE-19细胞后,通过噻唑兰(MTT)试验检测AF对ARPE-19细胞活力的影响;通过Western blot检测AF对表皮生长因子受体(EGFR)和磷酸化p38分裂原激活蛋白激酶(MAPK)及其下游蛋白表达的影响;采用免疫荧光染色检测EGFR的表达及变化。结果经2.0μMAF刺激12 h后,ARPE-19细胞胞体出现明显收缩、变形、细胞间隙扩大;MTT试验结果显示,2.0μMAF刺激12 h可抑制ARPE-19的细胞活力。Western blot结果检测显示,AF刺激可诱导总EGFR和MAPKAPK-2蛋白表达下调,同时可诱导p-EGFR、p-p38MAPK、p-MAPKAPK-2及p-HSP27蛋白表达上调。免疫荧光及鬼笔环肽染色检查结果显示,2.0μMAF刺激ARPE-19细胞12 h,可诱导EGFR从细胞膜转移到细胞质,细胞体收缩,细胞骨架破坏,F-肌动蛋白纤维变形,细胞核显著收缩。结论 AF可通过EGFR/p38MAPK信号通路抑制ARPE-19细胞的活力,其可能对增殖性玻璃体视网膜病变的发生、发展过程中RPE细胞异常存活具有潜在的防治作用。Objective To investigate the inhibitory mechanism of aureofin （AF） on cell viability of retinal pigmentepithelium （RPE）. Methods Human RPE cell strains （ARPE-19） were cultured in vitro and stimulated with 2.0 μM AF.The effect of AF on cell viability was determined by methyl thiazolyl tetrazolium （MTT） assay. Western blot was used todetect the effects of AF on epidermal growth factor receptor （EGFR）, phosphorylated p38 mitogen-activated protein kinase（MAPK） and the expression of their downstream proteins. Immunofluorescence staining was used to detect the expressionand change of EGFR. Results After 12 hours of stimulation with 2.0 μM AF, the cell body of ARPE-19 cells contracted,deformed and the cell gap enlarged significantly. MTT test results showed that the cell viability of ARPE-19 was inhibitedby 2.0 μM AF stimulation for 12 h. Western blot results showed that AF stimulation could induce down-regulation of totalEGFR and MAPKAPK-2 protein expression, and can induce up-regulation of p-EGFR, p-p38MAPK, p-MAPKAPK-2 andp -HSP27 protein expression. Immunofluorescence and phalloidin staining results showed that EGFR was induced totransfer from cell membrane to cytoplasm, cell body contraction, cytoskeleton destruction, F-actin fiber deformation, andnucleus significant contraction after 12 hours of stimulation with 2.0 μM AF for ARPE-19 cells. Conclusion AF caninhibit the cell viability of ARPE -19 cells through EGFR/p38MAPK signaling pathway, which may have potentialpreventive and therapeutic effects on the abnormal survival of RPE cells during the occurrence and development ofproliferative vitreoretinopathy.

关 键 词：金诺芬(AF) 视网膜色素上皮细胞(RPE) 表皮生长因子受体(EGFR) p38分裂原激活蛋白激酶(MAPK) 

分 类 号：R774.1[医药卫生—眼科]