柚皮素通过调控miR-29b的高表达对HepG2细胞胰岛素抵抗的改善作用  

Naringenin ameliorates insulin resistance in HepG2 cells by regulating high miR-29b expression

作　　者：王元 曾凯宏 余雪梅[1] 邓波[1] Wang Yuan;Zeng Kaihong;Yu Xuemei;Deng Bo(Dept of Clinical Nutrition,Sichuan Academy of Medical Sciences&Sichuan Provincial People′s Hospital,Chengdu 610072;School of Medicine,University of Electronic Science and Technology of China,Chengdu 610054;Dept of Health Management Center&Institute of Health Management,Sichuan Provincial People's Hospital,University of Electronic Science and Technology of China,Chengdu 610072)

机构地区：[1]四川省医学科学院•四川省人民医院临床营养科,成都610072 [2]电子科技大学医学院,成都610054 [3]四川省医学科学院•四川省人民医院健康管理研究所,成都610072

出　　处：《安徽医科大学学报》2024年第8期1423-1428,共6页Acta Universitatis Medicinalis Anhui

基　　金：四川省卫生和计划生育委员会科研课题(编号:150216);四川省中医药管理局中医药科研专项(编号:2021MS470);四川省人民医院院科研基金(编号:2023QN06)。

摘　　要：目的 探讨柚皮素(Nar)调控微小RNA-29b(miR-29b)的高表达对人肝癌细胞HepG2细胞胰岛素抵抗(IR)的作用,为研究Nar对糖尿病的防治作用机制做铺垫。方法 用100 nmol/L胰岛素刺激HepG2细胞建立HepG2细胞IR模型(IR-HepG2),分别用不同浓度(0、25、50、100μg/ml)的Nar干预IR-HepG2细胞;用葡萄糖试剂盒测定Nar对IR-HepG2细胞葡萄糖消耗的影响;对50μg/ml Nar干预组IR-HepG2细胞转染miR-29b mimic和miR-29b inhibitor,分别用实时定量反转录聚合酶链式反应(RT-qPCR)和蛋白质印迹法检测胰岛素信号传导通路中胰岛素受体底物-1(IRS-1)、蛋白激酶B/磷酸化蛋白激酶B(Akt/p-Akt)、葡萄糖转运子4(GLUT4)基因和蛋白的表达。结果 与IR-HepG2模型组比较,不同浓度Nar干预组细胞的葡萄糖消耗量升高(P<0.01),其中50μg/ml Nar干预IR组最明显(P<0.001),不同浓度Nar干预组IRS-1、Akt的mRNA表达增加(P<0.05),其中50μg/ml Nar干预组IRS-1、Akt的mRNA表达增加最明显(P<0.001),且50μg/ml Nar干预组GLUT4的mRNA表达增加(P<0.05),同时,不同浓度Nar干预组IRS-1、p-Akt的蛋白表达增加(P<0.001)。与IR-HepG2模型组相比,miR-29b mimic转染组细胞中IRS-1、Akt、GLUT4的mRNA表达和IRS-1、p-Akt蛋白表达降低(P<0.001),miR-29b inhibitor转染组中IRS-1、Akt、GLUT4的mRNA表达量和IRS-1、p-Akt蛋白表达无差异,Nar干预模型组和Nar干预转染miR-29b mimic组IRS-1、Akt、GLUT4的mRNA表达增加(P<0.001),IRS-1、p-Akt的蛋白表达增加(P<0.05)。与Nar干预模型组比较,Nar干预转染miR-29b mimic组IRS-1、Akt、GLUT4的mRNA表达无变化,IRS-1、p-Akt蛋白表达增加(P<0.05),Nar干预转染miR-29b inhibitor组IRS-1、Akt、GLUT4的mRNA表达和IRS-1、p-Akt的蛋白表达下降(P<0.001)。结论 Nar干预可增加IR-HepG2细胞葡萄糖的消耗,提高胰岛素抵抗HepG2细胞中IRS-1、Akt、GLUT4基因的表达量,增加IRS-1、p-Akt蛋白的表达量,Nar通过抑制miR-29b高表达增加IR-HepG2细胞IRS-1、p-Akt蛋白的表达,Objective To investigate the impact of naringenin(Nar)on insulin resistance(IR)in HepG2 cells and evaluate the role of mircoRNA-29b(miR-29b)expression in mediating this effect,thereby providing a foundation for further exploration into the mechanisms underlying naringenin's potential as a preventative and therapeutic agent for diabetes.Methods Insulin resistant HepG2(IR-HepG2)was established by stimulating HepG2 cells with 100 nmol/L insulin.Nar was treated with different concentrations(0,25,50,100μg/ml)in IR-HepG2 cells.The effect of Nar on glucose consumption in IR-HepG2 cells was determined with glucose kit.miR-29b mimic and inhibitor were transfected into IR-HepG2 cells of the 50μg/ml Nar intervention group,and the expressions of insulin receptor substrate-1(IRS-1),protein kinase B(Akt)/phosphorylated Akt(p-Akt),glucose transporter-4(GLUT4)genes and proteins in the insulin signaling pathway were detected by Real-time fluorescence quantitative reverse transcription polymerase chain reaction(RT-qPCR)and Western blot,respectively.Results Compared with IR-HepG2 model group,glucose consumption was increased in Nar intervention group with different concentrations(P<0.01),among which 50μg/ml Nar intervention group was the most significant(P<0.001),and mRNA expressions of IRS-1 and Akt were increased in Nar intervention group with different concentrations(P<0.05),the mRNA expression of IRS-1 and Akt in 50μg/ml Nar intervention group was the most significantly increased(P<0.001),and GLUT4 mRNA expression in 50μg/ml Nar intervention group was increased(P<0.05).The protein expressions of IRS-1 and p-Akt were increased in different Nar concentration groups(P<0.001).Compared with IR-HepG2 model group,mRNA expression of IRS-1,Akt and GLUT4 and protein expression of IRS-1 and p-Akt were decreased in miR-29b mimic transfected cells(P<0.001),mRNA expression of IRS-1,Akt and GLUT4 and protein expression of IRS-1 and p-Akt were not different in miR-29b inhibitor transfection group,Nar intervention model group and Nar inter

关 键 词：柚皮素 胰岛素抵抗 微小RNA-29b 

分 类 号：R151.3[医药卫生—营养与食品卫生学]