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目的利用双向电泳技术(2-DE)从蛋白质组学角度分析白藜芦醇对中波紫外线(UVB)照射的人角质形成细胞(HaCaT细胞)的保护作用,以探讨白藜芦醇防御UVB损伤的作用靶点及有关机制。方法 MTT法检测不同浓度的白藜芦醇对经或不经UVB照射的HaCaT细胞增殖的影响。2-DE实验分对照组、UVB组、白藜芦醇组、UVB+白藜芦醇干预组(共同干预组),提取各组蛋白进行2-DE实验,结合质谱(MALDI-TOF-MS)分析与数据库检索对差异蛋白质点进行鉴定。结果筛选10个存在明显差异的蛋白点,质谱鉴定出8种蛋白质,除4种功能未知蛋白外,其余4种蛋白分别为转录起始因子IIE-β(TFIIE-β)、组蛋白H1.2、锌指蛋白、MAGOH蛋白。UVB组细胞内组蛋白H1.2水平较其它各组明显增高(P<0.05);UVB组、UVB+白藜芦醇干预组的TFIIE-β及锌指蛋白表达水平均降低,UVB组下降最为明显(P<0.05);UVB组的MAGOH蛋白较各组明显减少(P<0.05)。结论应用2-DE联合质谱技术发现了白藜芦醇对UVB诱导的HaCaT细胞损伤防护作用相关的差异蛋白点。
Objective To investigate the protective effect of resveratrol on UVB-irradiated human keratinocytes (HaCaT cells) from the viewpoint of proteomics using two-dimensional electrophoresis (2-DE) to explore the protective effects of resveratrol against UVB damage. Targets and related mechanisms. Methods The effect of different concentrations of resveratrol on the proliferation of HaCaT cells irradiated with or without UVB was detected by MTT assay. The 2-DE experiment was divided into control group, UVB group, resveratrol group, UVB+resveratrol intervention group (co-intervention group), and 2-DE experiment was performed to extract each group of proteins and analyzed by mass spectrometry (MALDI-TOF-MS). Identifying differential protein spots with database searches. Results Ten protein spots with distinct differences were screened. Eight proteins were identified by mass spectrometry. In addition to the four unknown proteins, the other four proteins were the transcription initiation factor IIE-β (TFIIE-β) and histone H1.2. , zinc finger protein, MAGOH protein. The level of histone H1.2 in UVB group was significantly higher than that in other groups (P<0.05). The expression levels of TFIIE-β and zinc finger protein in UVB group and UVB+resveratrol intervention group were significantly lower than those in UVB group. (P<0.05); The MAGOH protein in UVB group was significantly lower than that in each group (P<0.05). Conclusion The differential protein spots related to UVB-induced HaCaT cell injury were detected by 2-DE combined with mass spectrometry.