【摘要】 目的:探讨复方水蛭滴眼液(SZ)防护人晶状体上皮细胞紫外线(UV)损伤的蛋白质组学机制,为将SZ作为防治白内障的有效药物提供实验依据。方法:采用SZ与人晶状体上皮细胞系HLE-B3细胞(HLEC)共同孵育,以吡喏克辛滴眼液(PS)作为阳性对照,经UV照射HLEC后,采用双向凝胶电泳(2-DE)和质谱(MS)技术,寻找HLEC有效的功能蛋白及SZ作用靶点。结果:①UV组与正常组比较:发现13个差异点。其中表达上调的差异点有5个,经质谱鉴定为:T复合蛋白1β亚基、核内不均一核糖核蛋白、磷酸甘油酸变位酶1、阻抑素(pro-hibitin)4种蛋白质;表达下调的差异点有8个,经质谱鉴定为:波形蛋白、半胱氨酰tRNA合成酶、锰超氧化物歧化酶3种蛋白质。②SZ组与UV组比较:发现12个差异蛋白点。其中表达上调的差异点8个,经质谱鉴定为:波形蛋白、半胱氨酰tRNA合成酶、锰超氧化物歧化酶3种蛋白质;表达下调的差异点4个,经质谱鉴定为:核内不均一核糖核蛋白、磷酸甘油酸变位酶1、角蛋白8Ⅱ和阻抑素4种蛋白质。结论:SZ可使UV照射引起HLEC下调的3种蛋白质(波形蛋白、半胱氨酰tRNA合成酶、锰超氧化物歧化酶)上调,又可使UV照射引起上调的4种蛋白质中的3种蛋白质下调(核内不均一核糖核蛋白、磷酸甘油酸变位酶1、阻抑素)。说明在SZ防护UV损伤中,以上6种蛋白质可能起重要作用。更多还原

【Abstract】 Objective:To investigation of proteomics mechanisms on compound Shui Zhi eye drop(SZ)protecting injury of human lens epithelia cells(HLEC)induced by ultraviolet(UV).This observation provides the experiment bases for clinical application of SZ on cataract.Methods:This work will use HLEC-B3 cells.The cultured HLEC were divided into control group;UV group;SZ group and pirenoxine solution(PS)group.The cells were exposed under UV.Then Using technology of two-dimensional electrophoresis(2-DE)and Mass Spectrometry(MS)to analysis for functional protein in HLEC induced by UV and target protein of SZ preventing injury of HLEC.Results:①UV group compared with control group:obtained different spots are 13,in which,5 spots regulated up.Four kinds of protein are identified by analysis of MS.These are T-complex protein 1 subunit beta,heterogeneous nuclear ribonucleoprotein,phosphoglycerate mutase 1 and prohibitin.Meanwhile,8 spots regulated down.Three kinds of protein are identified.These included vimentin,cysteinyl-tRNA synthetase and superoxide dismutase [Mn].②SZ group compared with UV group:8 spots regulated up,in which,three kinds of protein are identified.These are vimentin,cysteinyl-tRNA synthetase and superoxide dismutase [Mn].Meanwhile,4 spots regulated down,in which,four kinds of protein are identified.These are heterogeneous nuclear ribonucle-oprotein,phosphoglycerate mutase 1,keratin and prohibitin.Conclusion:SZ can regulate up expression of three kinds of protein that regulated down in UV irradiation.These are Vimentin,Cysteinyl-tRNA synthetase and Superoxide dismutase [Mn].Moreover,SZ can regulate down expression of three proteins that regulated up in UV irradiation.These are heterogeneous nuclear ribonucleoprotein,Phosphoglycerate mutase 1 and Prohibitin.This result show that above six kinds of protein can play important role in SZ preventing injury of HLEC induced by UV.`更多还原