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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。
Objective: To investigate the proteomics mechanism of compound leech eye drops (SZ) in protecting human lens epithelial cells from ultraviolet (UV) damage, and to provide experimental basis for SZ as an effective drug for the prevention and treatment of cataract. METHODS: SZ was co-incubated with human lens epithelial cell line HLE-B3 cells (HLEC) and pyridoxicin eye drops (PS) was used as a positive control. After UV irradiation, two-dimensional gel electrophoresis (2-DE) was performed. ) and mass spectrometry (MS) techniques to find HLEC effective functional proteins and SZ targets. Results: 1UV group compared with normal group: 13 differences were found. Among them, there were 5 differences in expression up-regulation, which were identified by mass spectrometry as: T complex protein 1β subunit, nuclear heterogeneous ribonucleoprotein, phosphoglycerate mutase 1, and pro-hibitin 4 proteins; There were 8 differences in down-regulation, identified by mass spectrometry as vimentin, cysteinyl tRNA synthetase, and manganese superoxide dismutase. 2SZ group compared with UV group: 12 differential protein spots were found. Among them, 8 were differentially expressed and identified by mass spectrometry: vimentin, cysteinyl tRNA synthetase, and manganese superoxide dismutase; 4 differences in down-regulation were identified by mass spectrometry: intranuclear Heterogeneous ribonucleoprotein, phosphoglycerate mutase 1, keratin 8II, and prohibitin 4 proteins. CONCLUSIONS: SZ can upregulate the three proteins (vimentin, cysteinyl tRNA synthetase, manganese superoxide dismutase) downregulated by HLEC caused by UV irradiation, and can also cause 3 of the 4 proteins upregulated by UV irradiation. Downregulation of proteins (nuclear heterogeneous ribonucleoprotein, phosphoglycerate mutase 1, repressin). This shows that the above six proteins may play an important role in SZ protection against UV damage.