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目的运用蛋白质组学技术分析维生素D3(VD3)对高糖负荷单核细胞的干预作用,探讨VD3在预防和治疗糖尿病时作用于单核免疫细胞的关键蛋白和分子机制。方法选择传代生长良好的U937细胞,经1,25-(OH)2-D3干预后进行高糖负荷培养,以未干预和单纯高糖负荷干预为对照组。提取细胞总蛋白后进行双向电泳,差异蛋白做质谱(MALDI-TOF-MS)鉴定。结果实验组与对照组比较,表达量变化1.5倍以上有30个蛋白质斑点,筛选其中6个差异蛋白点做MALDI-TOF-MS,成功鉴定出6种蛋白质,分别为:抑制素、70 kD的热休克蛋白8、过氧化氢酶、电子转移黄素蛋白、T-复合蛋白、磷酸丙糖异构酶。结论实验发现了VD3对高糖负荷U937细胞产生作用的相关的差异蛋白质,VD3可能通过降低氧化应激损伤和影响能量代谢的途径调节高糖负荷产生的损伤。
Objective To analyze the intervention of vitamin D3 (VD3) on high glucose-loaded monocytes by proteomic techniques and to explore the key proteins and molecular mechanisms of VD3 on mononuclear immune cells in the prevention and treatment of diabetes. Methods U937 cells, which were well-passaged and passaged, were selected and cultured with high glucose loading after 1,25- (OH) 2-D3 intervention. The intervention group without intervention and high glucose loading alone served as the control group. Total cellular protein was extracted and subjected to two-dimensional electrophoresis, and differential proteins were identified by MALDI-TOF-MS. Results Compared with the control group, there were 30 protein spots more than 1.5-fold change in the experimental group. Six protein spots were screened by MALDI-TOF-MS, and six proteins were identified, which were inhibin, 70 kD Heat shock protein 8, catalase, electron transfer flavoprotein, T-complex protein, triosephosphate isomerase. Conclusion VD3 can regulate the damage induced by high glucose load through decreasing the pathways of oxidative stress and energy metabolism, which are related to the effect of VD3 on U937 cells with high glucose.