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中国应用生理学杂志 ›› 2020, Vol. 36 ›› Issue (5): 503-508.doi: 10.12047/j.cjap.5984.2020.107

• 研究论文 • 上一篇    下一篇

微小核糖核酸106b对肝细胞葡萄糖异生的影响及机制*

付学东, 王寿懿, 朱志强, 邓幼平Δ   

  1. 武汉大学中南医院儿科, 湖北武汉 430071
  • 收稿日期:2019-12-03 修回日期:2020-07-24 发布日期:2021-02-25
  • 通讯作者: Tel: 027-67813138; E-mail: dengyp@whu.edu.cn
  • 基金资助:
    *湖北省自然科学基金资助项目(2018CFB560)

Effects of microRNA-106b on hepatic gluconeogenesis and its underlying mechanism

FU Xue-dong, WANG Shou-yi, ZHU Zhi-qiang, DENG You-pingΔ   

  1. Department of Pediatrics, Zhongnan Hospital of Wuhan University, Wuhan 430071, China
  • Received:2019-12-03 Revised:2020-07-24 Published:2021-02-25

摘要: 目的: 探讨微小核糖核酸106b(miR-106b)对肝细胞葡萄糖异生作用及其机制。方法: 正常人L02肝细胞培养于含10%胎牛血清的DMEM中,利用miR-106b模拟物和抑制剂(mimics和antagomiR,分别20 nmol/L)处理L02肝细胞24 h,Western blot法检测蛋白和磷酸化蛋白的表达,定量RT-PCR检测mRNA的表达,葡萄糖试剂盒检测培养液中葡萄糖含量。结果: miR-106b模拟物可明显增加磷酸烯醇式丙酮酸羧激酶(PEPCK)和葡萄糖-6-磷酸酶(G6Pase)的蛋白表达(P均<0.01)、增加磷酸烯醇式丙酮酸羧激酶1(PCK1)的mRNA表达(P<0.01)、降低葡萄糖激酶(GCK)的mRNA表达(P<0.01)。miR-106b抑制剂可显著降低PEPCK和G6Pase的蛋白表达(P均<0.01)、降低PCK1的mRNA表达(P<0.01)、增加GCK的mRNA表达(P<0.01)。此外,miR-106b模拟物或抑制剂可显著降低或增加信号转导和转录激活子3(STAT3)的蛋白表达(P均<0.01)。STAT3特异性抑制剂可显著拮抗miR-106b抑制剂对肝细胞葡萄糖异生的抑制作用。结论: miR-106b通过抑制STAT3信号通路而增加肝细胞葡萄糖异生。

关键词: 微小核糖核酸, 肝细胞, 葡萄糖异生, 信号转导和转录激活子3

Abstract: Objective: To investigate the potential effects of microRNA-106b (miR-106b) on gluconeogenesis in normal human liver cell line L02 and its underlying mechanisms. Methods: Normal human liver L02 cells were cultured in DMEM containing 10% FBS and transfected with 20 nmol/L of miR-106b mimic or antagomiR-106b, respectively. Twenty-four hours later after transfection, Western blot was performed to detect the levels of proteins or phosphorylated proteins. Quantitative RT-PCR was carried out to measure the mRNA expressions of gluconeogenesis-related genes. Glucose Assay Kit was used to detect the glucose contents in the medium. Results: MiR-106b mimic significantly increased the protein abundances of phosphoenolpyruvate carboxykinase (PEPCK) and glucose-6-phosphatase (G6Pase)(P<0.01,P<0.01, respectively), enhanced the mRNA expression of phosphoenolpyruvate carboxykinase 1 (PCK1) (P<0.01), and decreased the mRNA level of glucokinase (GCK) (P<0.01). AntagomiR-106b dramatically reduced the protein levels of PEPCK and G6Pase (P<0.01,P<0.01, respectively), suppressed the mRNA level of PCK1 (P<0.01), and increased the mRNA level of GCK(P<0.01). In addition,miR-106b mimic or antagomiR-106b significantly reduced or enhanced the protein levels of signal transducer and activator of transcription 3 (STAT3)(P<0.01,P<0.01, respectively).The inhibition of STAT3 by its specific inhibitor abolished the inhibitory effects of antagomiR-106b on hepatic gluconeogenesis. Conclusion: miR-106b increases hepatic gluconeogenesis by inhibiting the STAT3 signaling pathway.

Key words: MicroRNA-106b, gluconeogenesis, STAT3, hepatocytes

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