摘要
目的探讨miR-93-5p对胰岛素抵抗细胞模型中肝细胞生长因子(hepatocyte growth factor,HGF)、葡萄糖转运蛋白4(glucose transporter 4,GLUT4)表达及GLUT4分布的影响。方法将细胞分为对照组、模型组、miR-93-5p inhibitor组、inhibitor-NC组、miR-93-5p mimics组和mimics-NC组。对照组细胞正常培养,模型组构建胰岛素抵抗细胞模型,其余组分别转染miR-93-5p inhibitor、inhibitor-NC、miR-93-5p mimics、mimics-NC,转染完成后构建胰岛素抵抗模型。CCK-8检测细胞存活率;试剂盒检测细胞上清液中葡萄糖含量,计算葡萄糖消耗量;qPCR检测miR-93-5p、HGF、GLUT4的mRNA表达水平;Western blot检测HGF、GLUT4蛋白表达水平;免疫荧光检测GLUT4的表达和分布。结果与对照组相比,模型组细胞存活率、葡萄糖消耗量、HGF、GLUT4 mRNA和蛋白表达水平均显著降低(均P<0.01),miR-93-5p水平显著升高(P<0.01),GLUT4膜分布减少。与模型组相比,miR-93-5p inhibitor组细胞存活率、葡萄糖消耗量、HGF、GLUT4 mRNA和蛋白表达水平均显著升高(均P<0.05),miR-93-5p水平显著降低(P<0.01),GLUT4膜分布增多;mimics组结果相反。结论miR-93-5p能够抑制胰岛素抵抗细胞模型中HGF和GLUT4的表达,并减少GLUT4的膜分布,具有促进胰岛素抵抗的功能。
Objective To investigate the effect of miR-93-5p on the expression of hepatocyte growth factor(HGF),glucose transporter 4(GLUT4)and distribution of GLUT4 in insulin resistance cell model.Methods The cells were divided into control group,model group,miR-93-5p inhibitor group,inhibitor-NC group,miR-93-5p mimics group and mimics-NC group.The cells in control group were cultured normally,while insulin resistance cell model was constructed in cells of model group.The other groups were transfected with miR-93-5p inhibitor,inhibitor-NC,miR-93-5p mimics and mimics-NC,respectively.And the insulin resistance model was constructed after transfection.Cell viability was detected by CCK-8.The kit was used to measure the glucose content in the cell supernatant and the glucose consumption was calculated.The expressions of miR-93-5p,HGF and GLUT4 mRNA were detected by qPCR.The expressions of HGF and GLUT4 protein were detected by Western blotting.The expression and distribution of GLUT4 were detected by immunofluorescence.Results Compared with the control group,the cell survival rate,glucose consumption,HGF,GLUT4 mRNA and protein expression levels were significantly decreased(all P<0.01),miR-93-5p level was significantly increased(P<0.01),and GLUT4 membrane distribution was decreased in the model group.Compared with the model group,the cell survival rate,glucose consumption,HGF,GLUT4 mRNA and protein expression levels were significantly increased(all P<0.05),miR-93-5p level was significantly decreased(P<0.01),and GLUT4 membrane distribution was increased in the miR-93-5p inhibitor group.The results were opposite in the miR-93-5p mimics group.Conclusion miR-93-5p can inhibit the expression of HGF and GLUT4 and reduce the membrane distribution of GLUT4 in insulin resistance cell model,which can promote insulin resistance.
作者
周曼
吴军
干定云
陈婉
曹萍
李广利
侯以琳
Zhou Man;Wu Jun;Gan Dingyun(Department of Endocrinology,Wuhan Third Hospital(Tongren Hospital of Wuhan University),Wuhan 430060,China)
出处
《华中科技大学学报（医学版）》
CAS
CSCD
北大核心
2023年第6期823-827,共5页
Acta Medicinae Universitatis Scientiae et Technologiae Huazhong
基金
武汉市卫生健康委员会医学科研项目(No.WX21D17)。
