维生素D_3干预高糖负荷U937细胞的差异蛋白质组分析  

Proteomic differential display analysis of high glucose load human U937 cell line stimulated by vitamin D_3

作　　者：孙双玲 马爱国[1] 张秀莲[1] 汪求真[1] 姜颖[1] 

机构地区：[1]青岛大学医学院,青岛266021

出　　处：《卫生研究》2014年第3期366-371,共6页Journal of Hygiene Research

基　　金：国家自然科学研究基金(No.81172662);2011年达能营养宣教研究基金

摘　　要：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 The purpose of this study was to analysis on the affection of key protein expression by vitamin D3 , and to investigate the mechanism of vitamin D3 on contributing to diabetes. Method Good status U937 cells were cultured by high glucose medium which contained 45mmol/L D-Glucose, and treated by 1, 25-（ OH）2-D3. The control groups were U937 cells cultured by high glucose. The whole cell proteins were extracted and separated by 2-dimentional gel electrophoresis （2-DE）, differential expression proteins were identified by matrix-assisted laser desorption/ionization-time of flight-mass spectrometry （MALDI-TOF-MS）. Result There were 30 protein spots whose expression was significantly different between vitamin D-treated and untreated high glucose loaded cells. Six protein spots were identified by MALDI-TOF-MS, three of which, prohibitin, heat shock 70 kD protein 8, and catalase were protection of cell stress proteins, one of which, electron transport flavoprotein, was respiratory chain related proteins, two of which, fumarylacetoacetate hydrolase and triosephosphate isomerase, were cell metabolism related proteins. Conclusion The differentially expressed proteins of high glucose load human U937 cell line stimulated by vitamin D3 were found. These differential proteins were mainly related to oxidative stress and energy metabolism. These data suggested that 1, 25-（ OH）E-D3 might regulate cell high glucose load by reduction oxidative stress injury and affection energy metabolism.

关 键 词：维生素D3 高糖 U937 蛋白质组 氧化应激 

分 类 号：Q565.3[生物学—生物化学] R587.1[医药卫生—内分泌]