siRNA干扰MafA对小鼠胰岛β细胞的生长以及胰岛素相关基因的表达分析

Effects of siRNA interference with MafA on the growth of mouse isletβcells and the expression of insulin-related genes

目的:探讨肌腱膜纤维肉瘤癌基因同源物A(MafA)对小鼠胰岛β细胞Min6的增殖以及胰岛素相关基因mRNA及蛋白表达的影响。方法:实验分为MafA siRNA转染的MafA干扰A组、MafA干扰B组、MafA干扰C组及未转染的对照D组、对照E组、对照F组。MafA干扰A组和对照D组培养的葡萄糖浓度为5.6 mmol/L,MafA干扰B组和对照E组为10 mmol/L,MafA干扰C组和对照F组为25 mmol/L。采用实时荧光定量PCR及Western blotting检测mRNA及蛋白表达,MTT法检测胰岛细胞增殖,ELISA测定胰岛素浓度。结果:在MaFA mRNA及蛋白的表达水平,MafA干扰B组和对照E组、MafA干扰C组、对照F组相比差异均有统计学意义(P<0.05)。MafA干扰C组Min6细胞增殖水平低于对照F组(P<0.05)。MafA干扰C组Insulin-1、Insulin-2 mRNA相对表达水平均低于对照F组(P<0.05)。MafA干扰C组胰岛素浓度低于对照F组(P<0.05)。结论:MafA基因与小鼠胰岛β细胞增殖和胰岛素分泌相关,其作用可能与血糖浓度有关。

Objective:To investigate the effect of v-maf musculoaponeurotic fibrosarcoma oncogene homologue A(MafA)on the proliferation of mouse isletβcells Min6,and the mRNA and protein expression of insulin-related genes.Methods:The experiment was divided into MafA siRNA transfected MafA interference group A,MafA interference group B,MafA interference group C,and non-transfected control group D,control group E,control group F.MafA interference group A and control group D were treated with 5.6 mmol/L glucose,MafA interference group B and control group E were treated with 10 mmol/L glucose,and MafA interference group C and control group F were treated with 25 mmol/L glucose.The expression of mRNA and protein was detected by real-time PCR and Western blotting.The proliferation of islet cells was detected by MTT assay.The concentration of insulin was determined by ELISA.Results:There were significant differences in the expression of MafA mRNA and protein between MafA interference group B and control group E,MafA interference group C and control group F(P<0.05).The proliferation level of Min6 cells in MafA interference group C was lower than that in control group F(P<0.05).The relative expression levels of Insulin-1 and Insulin-2 mRNA in MafA interference group C were lower than those in control group F(P<0.05).The insulin concentration in MafA interference group C was lower than that in control group F(P<0.05).Conclusions:MafA gene is related to the proliferation and insulin secretion of mouse isletβcells,which may be related to the concentration of blood glucose.