钙结合蛋白S100A16对胰岛素抵抗的作用研究

Effects of calcium binding protein S100A16 on promoting insulin resistance

目的探究钙结合蛋白S100A16在胰岛素抵抗中的作用。方法用S100A16抗体进行免疫沉淀,然后用蛋白质谱分析寻找与S100A16相互作用的蛋白。实验1转染Vector质粒的HepG2细胞作为对照,用转染shRNA质粒、S100A16过表达质粒干预作为处理组。实验2以慢性胰岛素刺激细胞构建胰岛素抵抗模型,采用转染shRNA质粒的细胞作为对照,用未转染和转染Vector质粒干预作为处理组。实验3以不做任何处理的细胞作为对照,在胰岛素抵抗模型中用吡格列酮干预作为处理组。Western blot检测相关蛋白的表达水平。组间比较采用成组t检验。结果与转染Vector质粒比较,转染S100A16过表达质粒中胎球蛋白A表达(1.39±0.54比2.85±0.25)水平上调(P<0.05);与转染Vector质粒比较,转染shRNA质粒胎球蛋白A蛋白表达(0.36±0.03比0.20±0.03)水平降低(P<0.01)。在胰岛素抵抗条件下,与转染shRNA质粒的细胞比较,未转染和转染Vector质粒的IRS-2蛋白表达(0.11±0.04比1.65±0.48)水平上调(P<0.01);与不做任何处理的细胞比较,用吡格列酮处理的细胞IRS-2表达(0.26±0.11比0.52±0.05)水平上升(P<0.01)。结论S100A16在HepG2细胞中通过胎球蛋白A促进胰岛素抵抗。

Objective To explore the effects of S100A16,a calcium-binding protein on insulin resistance.Methods Immunoprecipitation was performed with S100A16 antibody,and mass spectrometry was used to find proteins interacting with S100A16.HepG2 cells transfected with shRNA plasmids or S100A16 overexpression plasmids were defined as experimental group 1 and HepG2 cells transfected with vector plasmids as control group 1.Chronic insulin stimulation was done to the cells to establish insulin resistance models,in which cells transfected with shRNA plasmids were experimental group 2.Those not transfected or transfected with vector plasmids were set to be control group 2.Cells treated with pioglitazone were experimental group 3 and those without pioglitazone treatment were control group 3 in insulin resistance group.The expression levels of relevant proteins were detected by Western blot.Unpaired t test was used for statistical analysis.Results Compared with control group 1,the expression of Fetuin A in overexpressed S100A16 group in experimental group 1 was increased significantly(1.39±0.54 vs 2.85±0.25,P<0.05),while the expression of Fetuin A in shRNA group in experimental group 1 was significantly decrease(0.36±0.03 vs 0.20±0.03,P<0.01).In contrast to control group 2,the expression of IRS-2(insulin receptor substrate-2)in experimental group 2 was significantly increased under the condition of insulin resistance(0.11±0.04 vs 1.65±0.48,P<0.01).And compared with control group 3,the expression of IRS-2 in experimental group 3 was obviously increased(0.26±0.11 vs 0.52±0.05,P<0.01).Conclusion S100A16 promotes insulin resistancein HepG2 cells via Fetuin A.