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

孙志兵; 杨娟

doi:10.13898/j.cnki.issn.1000-2200.2022.11.002

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

孙志兵,
杨娟

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

摘要

摘要:
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。

Abstract:
ObjectiveTo investigate the effect of v-maf musculoaponeurotic fibrosarcoma oncogene homologue A(MafA) on the proliferation of mouse islet β cells Min6, and the mRNA and protein expression of insulin-related genes.
MethodsThe 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.
ResultsThere 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).
ConclusionsMafA gene is related to the proliferation and insulin secretion of mouse islet β cells, which may be related to the concentration of blood glucose.

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