吉林大学学报(医学版) ›› 2018, Vol. 44 ›› Issue (01): 36-40.doi: 10.13481/j.1671-587x.20180107

• 基础研究 • 上一篇    下一篇

Exendin-4对胰岛素抵抗人肝癌HepG2细胞脂代谢相关因子基因表达的影响

苏寒1,2, 张美家1,2, 王怀杰1,2, 李娜1,2, 霍连广1,2, 李桂芝3, 戴功1,2, 高志芹1,4, 杨晓云1,3, 曲梅花1,2   

  1. 1. 潍坊医学院山东省高校应用药理学重点实验室, 山东 潍坊 261053;
    2. 潍坊医学院药学院药理学教研室, 山东 潍坊 261053;
    3. 潍坊医学院临床医学院生物化学教研室, 山东 潍坊 261053;
    4. 潍坊医学院 生物科学与技术学院细胞生物学教研室, 山东 潍坊 261053
  • 收稿日期:2017-09-27 出版日期:2018-01-28 发布日期:2018-01-24
  • 通讯作者: 曲梅花,教授,硕士研究生导师(Tel:0536-8462466,E-mail:qumeihua2016@163.com);杨晓云,副教授,硕士研究生导师(Tel:0536-8462465,E-mail:xiaoyuny1118@163.com) E-mail:qumeihua2016@163.com;xiaoyuny1118@163.com
  • 作者简介:苏寒(1992-),女,山东省济宁市人,在读医学硕士,主要从事内分泌和代谢性疾病方面的研究。
  • 基金资助:
    国家自然科学基金资助课题(31671208,H1304);山东省科技厅自然科学基金资助课题(ZR2015HL128,ZR2009CL047,ZR2015HL056);山东省教育厅高等学校科技计划基金资助课题(J14LK15,J15LM10)

Effects of Exendin-4 on expressions of lipid metabolism related genes in HepG2 cells with insulin resistance

SU Han1,2, ZHANG Meijia1,2, WANG Huaijie1,2, LI Na1,2, HUO Lianguang1,2, LI Guizhi3, DAI Gong1,2, GAO Zhiqin1,4, YANG Xiaoyun1,3, QU Meihua1,2   

  1. 1. Department of Pharmacology, Key Laboratory of Applied Pharmacology in Universities of Shandong Province, Weifang Medical University, Weifang 261053, China;
    2. Department of Pharmacology, School of Pharmacy, Weifang Medical University, Weifang 261053, China;
    3. Department of Biochemistry, Weifang Medical University, Weifang 261053, China;
    4. School of Biological Science and Technology, Weifang Medical University Department of Biology, Weifang 261053, China
  • Received:2017-09-27 Online:2018-01-28 Published:2018-01-24

摘要: 目的:探讨Exendin-4(Ex-4)对胰岛素抵抗(IR)人肝癌HepG2细胞脂代谢相关因子表达的影响,阐明Ex-4改善IR的作用。方法:选取处于对数生长期的人肝癌HepG2细胞,采用高浓度胰岛素诱导HepG2细胞制备IR模型(HepG2-IR细胞),再将其分为对照组(不含胰岛素的HepG2细胞)、IR组(IR模型,即HepG2-IR细胞)和Ex-4组(IR模型中加入10 nmol·L-1 Ex-4)。采用葡萄糖氧化酶法(GOD-POD)检测细胞中葡萄糖消耗量,采用油红O染色观察细胞形态及胞内脂滴形成情况,应用甘油三酯(TG)试剂盒检测细胞中TG水平,采用荧光定量PCR(qRT-PCR)法检测细胞中乙酰辅酶A羧化酶(ACC)、脂肪酸合成酶(FAS)、固醇调节元件结合蛋白1c(SREBP-1c)和载脂蛋白B100(apoB100)等脂代谢相关因子mRNA表达水平。结果:HepG2细胞建立IR模型后,与对照组比较,IR组HepG2-IR细胞葡萄糖消耗量明显降低(P<0.01);与IR组比较,Ex-4组HepG2-IR细胞葡萄糖消耗量明显增加(P<0.05)。油红O染色法,与对照组比较,IR组细胞含脂率明显升高(P<0.05);与IR组比较,Ex-4组细胞中含脂率明显降低(P<0.05)。与对照组比较,IR组细胞中TG水平明显升高(P<0.01);与IR组比较,Ex-4组细胞中TG水平明显降低(P<0.05)。qRT-PCR检测,与对照组比较,IR组细胞中ACC、FAS和SREBP-1c mRNA表达水平明显升高(P<0.01),apoB100mRNA表达水平明显降低(P<0.05);与IR组比较,Ex-4组细胞中ACC、FAS和SREBP-1c mRNA表达水平降低(P<0.05),apoB100 mRNA表达水平升高(P<0.01)。结论:Ex-4可通过调节人肝癌HepG2细胞脂类代谢相关因子的表达进而改善IR。

关键词: Exendin-4, 胰岛素抵抗, 乙酰辅酶A羧化酶, 脂代谢, 脂肪酸合成酶, HepG2细胞

Abstract: Objective: To investigate the effects of Exendin-4 (Ex-4) on the expressions of lipid metabolism related genes in the human liver cancer HepG2 cells with insulin resistance (IR), and to elucidate the effect of Ex-4 in improvement of IR. Methods: The HepG2 cells in logarithmic growth phase were induced into IR model with high concentration of insulin, then divided into control group (HepG2 cells), IR group (HepG2 cells were treated with insulin,HepG2-IR cells), and Ex-4 group (HepG2-IR cells were treated with Ex-4). Glucose oxidase (GOD-POD) kit was used to detect the consumption of glucose.The cell morphology and intracellular lipid drip formation were observed by Oil red O staining. The triglyceride (TG) level in cells was detected by kit; qRT-PCR was used to detect the mRNA expression levels of acetyl-CoA carboxylase (ACC), fatty acid synthase (FAS), sterol regulatory element-binding protein-1c (SREBP-1c) and apolipoprotein B100 (apoB100). Results: Compared with control group(HepG2 cells), the glucose consumption in the HepG2-IR cells in IR group was significantly decreased (P<0.01). Compared with IR group, the glucose consumption in the HepG2-IR cells in Ex-4 group was increased (P<0.05). The Oil O red staining results showed that compared with control group, the fat percentage in the HepG2-IR cells in IR group was increased(P<0.05); compared with IR group, the fat percentage in Ex-4 group was decreased (P<0.05). Compared with control group, the level of TG in the cells in IR group was significantly increased (P<0.01); compared with IR group, the level of TG in the cells in Ex-4 group was significantly decreased (P<0.05).The qT-PCR results showed that compared with control group, the expression levels of ACC FAS and SREBP-1c mRNA in the cells in IR group were increased (P<0.01), and the expression level of apoB100 mRNA was decreased (P<0.05); compared with IR group, the expression levels of ACC, FAS and SREBP-1c mRNA in the cells in Ex-4 group were decreased (P<0.05),and the expression level of apoB100 mRNA was increased (P<0.01). Conclusion: Ex-4 can regulate the expressions of lipid metabolism related genes in the HepG2 cells and improve IR.

Key words: HepG2 cells, insulin resistance, lipid metabolism, Exendin-4, acetyl-CoA carboxylase, fatty acid synthase

中图分类号: 

  • R735.7