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Upregulation of miR-125b by estrogen protects against non-alcoholic fatty liver in female mice

  • Zhi-Chun Zhang
    Affiliations
    Key Laboratory of Metabolism and Molecular Medicine, Ministry of Education, Department of Biochemistry and Molecular Biology, Fudan University Shanghai Medical College, Shanghai 200032, PR China
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  • Yan Liu
    Affiliations
    Key Laboratory of Metabolism and Molecular Medicine, Ministry of Education, Department of Biochemistry and Molecular Biology, Fudan University Shanghai Medical College, Shanghai 200032, PR China
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  • Liu-Ling Xiao
    Affiliations
    Key Laboratory of Metabolism and Molecular Medicine, Ministry of Education, Department of Biochemistry and Molecular Biology, Fudan University Shanghai Medical College, Shanghai 200032, PR China
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  • Shu-Fen Li
    Affiliations
    Key Laboratory of Metabolism and Molecular Medicine, Ministry of Education, Department of Biochemistry and Molecular Biology, Fudan University Shanghai Medical College, Shanghai 200032, PR China
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  • Jing-Hui Jiang
    Affiliations
    Key Laboratory of Metabolism and Molecular Medicine, Ministry of Education, Department of Biochemistry and Molecular Biology, Fudan University Shanghai Medical College, Shanghai 200032, PR China
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  • Yue Zhao
    Affiliations
    Key Laboratory of Metabolism and Molecular Medicine, Ministry of Education, Department of Biochemistry and Molecular Biology, Fudan University Shanghai Medical College, Shanghai 200032, PR China
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  • Shu-Wen Qian
    Affiliations
    Key Laboratory of Metabolism and Molecular Medicine, Ministry of Education, Department of Biochemistry and Molecular Biology, Fudan University Shanghai Medical College, Shanghai 200032, PR China
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  • Qi-Qun Tang
    Affiliations
    Key Laboratory of Metabolism and Molecular Medicine, Ministry of Education, Department of Biochemistry and Molecular Biology, Fudan University Shanghai Medical College, Shanghai 200032, PR China

    Institute of Stem Cell and Regenerative Medicine, Institutes of Biomedical Sciences, Fudan University, Shanghai 200032, PR China
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  • Xi Li
    Correspondence
    Corresponding author. Address: 138 Yi Xue Yuan Road, P.O. Box 238, Fudan University Shanghai Medical College, Shanghai 200032, PR China. Tel./fax: +86 21 54237290.
    Affiliations
    Key Laboratory of Metabolism and Molecular Medicine, Ministry of Education, Department of Biochemistry and Molecular Biology, Fudan University Shanghai Medical College, Shanghai 200032, PR China
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Published:August 10, 2015DOI:https://doi.org/10.1016/j.jhep.2015.07.037

      Background & Aims

      Due to the protective effect of estrogen against hepatic fat accumulation, the prevalence of non-alcoholic fatty liver disease (NAFLD) in premenopausal women is lower than that in men at the same age and in postmenopausal women. Our study was to further elucidate an underlying mechanism by which estrogen prevents NAFLD from miRNA perspective in female mice.

      Methods

      miRNA expression was evaluated by TaqMan miRNA assay. Luciferase and ChIP assay were done to validate regulation of miR-125b by estrogen via estrogen receptor alpha (ERα). Nile red and Oil red O staining were used to check lipid content. Overexpressing or inhibiting the physiological role of miR-125b in the liver of mice through injecting adenovirus were used to identify the function of miR-125b in vivo.

      Results

      miR-125b expression was activated by estrogen via ERα in vitro and in vivo. miR-125b inhibited lipid accumulation both in HepG2 cells and primary mouse hepatocytes. Consistently, ovariectomized or liver-specific ERα knockdown mice treated with miR-125b overexpressing adenoviruses were resistant to hepatic steatosis induced by high-fat diet, due to decreased fatty acid uptake and synthesis and decreased triglyceride synthesis. Conversely, inhibiting the physiological role of miR-125b with a sponge decoy slightly promoted liver steatosis with a high-fat diet. Notably, we provided evidence showing that fatty acid synthase was a functional target of miR-125b.

      Conclusion

      Our findings identify a novel mechanism by which estrogen protects against hepatic steatosis in female mice via upregulating miR-125b expression.

      Graphical abstract

      Abbreviations:

      NAFLD (non-alcoholic fatty liver disease), ChIP (chromatin immunoprecipitation), HFD (high-fat diet), ALT (alanine aminotransferase), AST (aspartate aminotransferase), FFA (free fatty acid), FAS (fatty acid synthase), SCD1 (stearoyl coenzyme A desaturase 1), SREBP-1C (sterol regulatory element binding protein 1C), ACC (acetyl coenzyme A carboxylase), PPAR (peroxisome proliferator-activated receptor), LXRα (liver X receptor alpha), DGAT (diacylglycerol acyl coenzyme A acyltransferase), CPT1 (carnitine palmitoyltransferase 1), MCD (malonyl coenzyme A decarboxylase), LCAD (long-chain acyl coenzyme A dehydrogenase), UCP2 (uncoupling protein 2), NC (negative control), WAT (white adipose tissue), TG (triglyceride), ERE (estrogen response element)

      Keywords

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      References

      Author names in bold designate shared co-first authorship

        • Angulo P.
        Nonalcoholic fatty liver disease.
        N Engl J Med. 2002; 346: 1221-1231
        • De Alwis N.M.
        • Day C.P.
        Non-alcoholic fatty liver disease: the mist gradually clears.
        J Hepatol. 2008; 48: S104-S112
        • Starley B.Q.
        • Calcagno C.J.
        • Harrison S.A.
        Nonalcoholic fatty liver disease and hepatocellular carcinoma: a weighty connection.
        Hepatology. 2010; 51: 1820-1832
        • Anstee Q.M.
        • Targher G.
        • Day C.P.
        Progression of NAFLD to diabetes mellitus, cardiovascular disease or cirrhosis.
        Nat Rev Gastroenterol Hepatol. 2013; 10: 330-344
        • Suzuki A.
        • Abdelmalek M.F.
        Nonalcoholic fatty liver disease in women.
        Women’s Health. 2009; 5: 191-203
        • Yang J.D.
        • Abdelmalek M.F.
        • Pang H.
        • Guy C.D.
        • Smith A.D.
        • Diehl A.M.
        • et al.
        Gender and menopause impact severity of fibrosis among patients with nonalcoholic steatohepatitis.
        Hepatology. 2014; 59: 1406-1414
        • Zhu L.
        • Brown W.C.
        • Cai Q.
        • Krust A.
        • Chambon P.
        • McGuinness O.P.
        • et al.
        Estrogen treatment after ovariectomy protects against fatty liver and may improve pathway-selective insulin resistance.
        Diabetes. 2013; 62: 424-434
        • Jones M.E.E.
        • Thorburn A.W.
        • Britt K.L.
        • Hewitt K.N.
        • Wreford N.G.
        • Proietto J.
        • et al.
        Aromatase-deficient (ArKO) mice have a phenotype of increased adiposity.
        Proc Natl Acad Sci U S A. 2000; 97: 12735-12740
        • Nguyen M.C.
        • Stewart R.B.
        • Banerji M.A.
        • Gordon D.H.
        • Kral J.G.
        Relationships between tamoxifen use, liver fat and body fat distribution in women with breast cancer.
        Int J Obes Relat Metab Disord. 2001; 25: 296-298
        • Cole L.K.
        • Jacobs R.L.
        • Vance D.E.
        Tamoxifen induces triacylglycerol accumulation in the mouse liver by activation of fatty acid synthesis.
        Hepatology. 2010; 52: 1258-1265
        • Nemoto Y.
        • Toda K.
        • Ono M.
        • Kiyomi F.A.
        • Saibara T.
        • Onishi S.
        • et al.
        Altered expression of fatty acid–metabolizing enzymes in aromatase-deficient mice.
        J Clin Invest. 2000; 105: 1819-1825
        • Esau C.
        • Davis S.
        • Murray S.F.
        • Yu X.X.
        • Pandey S.K.
        • Pear M.
        • et al.
        MiR-122 regulation of lipid metabolism revealed by in vivo antisense targeting.
        Cell Metab. 2006; 3: 87-98
        • Dávalos A.
        • Goedeke L.
        • Smibert P.
        • Ramírez C.M.
        • Warrier N.P.
        • Andreo U.
        MiR-33a/b contribute to the regulation of fatty acid metabolism and insulin signaling.
        Proc Natl Acad Sci U S A. 2011; 108: 9232-9237
        • Ng R.
        • Wu H.
        • Xiao H.
        • Chen X.
        • Willenbring H.
        • Steer C.J.
        • et al.
        Inhibition of mir-24 expression in liver prevents hepatic lipid accumulation and hyperlipidemia.
        Hepatology. 2014; 60: 554-564
        • Xiao F.
        • Yu J.
        • Liu B.
        • Guo Y.
        • Li K.
        • Deng J.
        • et al.
        A novel function of microRNA 130a–3p in hepatic insulin sensitivity and liver steatosis.
        Diabetes. 2014; 63: 2631-2642
        • Guan Y.
        • Yao H.
        • Zheng Z.
        • Qiu G.
        • Sun K.
        MiR-125b targets BCL3 and suppresses ovarian cancer proliferation.
        Int J Cancer. 2011; 128: 2274-2283
        • Iorio M.V.
        • Ferracin M.
        • Liu C.G.
        • Veronese A.
        • Spizzo R.
        • Sabbioni S.
        • et al.
        MicroRNA gene expression deregulation in human breast cancer.
        Cancer Res. 2005; 65: 7065-7070
        • Alpini G.
        • Glaser S.S.
        • Zhang J.P.
        • Francis H.
        • Han Y.
        • Gong J.
        • et al.
        Regulation of placenta growth factor by microRNA-125b in hepatocellular cancer.
        J Hepatol. 2011; 55: 1339-1345
        • Tili E.
        • Michaille J.J.
        • Luo Z.
        • Volinia S.
        • Rassenti L.Z.
        • Kipps T.J.
        • et al.
        The down-regulation of miR-125b in chronic lymphocytic leukemias leads to metabolic adaptation of cells to a transformed state.
        Blood. 2012; 120: 2631-2638
        • Donadeu F.X.
        • Schauer S.N.
        • Sontakke S.D.
        Involvement of miRNAs in ovarian follicular and luteal development.
        J Endocrinol. 2012; 215: 323-334
        • Mishima T.
        • Takizawa T.
        • Luo S.S.
        • Ishibashi O.
        • Kawahigashi Y.
        • Mizuguchi Y.
        • et al.
        MicroRNA (miRNA) cloning analysis reveals sex differences in miRNA expression profiles between adult mouse testis and ovary.
        Reproduction. 2008; 136: 811-822
        • Murphy A.J.
        • Guyre P.M.
        • Pioli P.A.
        Estradiol suppresses NF-κB activation through coordinated regulation of let-7a and miR-125b in primary human macrophages.
        J Immunol. 2010; 184: 5029-5037
        • Li S.
        • Chen X.
        • Zhang H.
        • Liang X.
        • Xiang Y.
        • Yu C.
        • et al.
        Differential expression of microRNAs in mouse liver under aberrant energy metabolic status.
        J Lipid Res. 2009; 50: 1756-1765
        • Han S.I.
        • Komatsu Y.
        • Murayama A.
        • Steffensen K.R.
        • Nakagawa Y.
        • Nakajima Y.
        • et al.
        Estrogen receptor ligands ameliorate fatty liver through a nonclassical estrogen receptor/Liver X receptor pathway in mice.
        Hepatology. 2014; 59: 1791-1802
        • Paquette A.
        • Wang D.
        • Jankowski M.
        • Gutkowska J.
        • Lavoie J.M.
        Effects of ovariectomy on PPAR alpha, SREBP-1c, and SCD-1 gene expression in the rat liver.
        Menopause. 2008; 15: 1169-1175
        • Castellano L.
        • Giamas G.
        • Jacob J.
        • Coombes R.C.
        • Lucchesi W.
        • Thiruchelvam P.
        • et al.
        The estrogen receptor-alpha-induced microRNA signature regulates itself and its transcriptional response.
        Proc Natl Acad Sci U S A. 2009; 106: 15732-15737
        • Di Leva G.
        • Piovan C.
        • Gasparini P.
        • Ngankeu A.
        • Taccioli C.
        • Briskin D.
        • et al.
        Estrogen mediated-activation of miR-191/425 cluster modulates tumorigenicity of breast cancer cells depending on estrogen receptor status.
        PLoS Genet. 2013; 9: e1003311
        • Kilnge C.M.
        Estrogen receptor interaction with estrogen response elements.
        Nucleic Acids Res. 2001; 29: 2905-2919
        • Bryzgalova G.
        • Lundholm L.
        • Portwood N.
        • Gustafsson J.A.
        • Khan A.
        • Efendic S.
        • et al.
        Mechanisms of antidiabetogenic and body weight-lowering effects of estrogen in high-fat diet-fed mice.
        Am J Physiol Endocrinol Metab. 2008; 295: E904-E912
        • Yoshizawa T.
        • Karim M.F.
        • Sato Y.
        • Senokuchi T.
        • Miyata K.
        • Fukuda T.
        • et al.
        SIRT7 controls hepatic lipid metabolism by regulating the ubiquitin-proteasome pathway.
        Cell Metab. 2014; 19: 712-721
        • Kim J.K.
        • Noh J.H.
        • Jung K.H.
        • Eun J.W.
        • Bae H.J.
        • Kim M.G.
        • et al.
        Sirtuin7 oncogenic potential in human hepatocellular carcinoma and its regulation by the tumor suppressors MiR-125a-5p and MiR-125b.
        Hepatology. 2013; 57: 1055-1067
        • Han Y.
        • Liu Y.
        • Zhang H.
        • Wang T.
        • Diao R.
        • Jiang Z.
        • et al.
        Hsa-miR-125b suppresses bladder cancer development by down-regulating oncogene SIRT7 and oncogenic long non-coding RNA MALAT1.
        FEBS Letters. 2013; 587: 3875-3882
        • Shin J.
        • He M.
        • Liu Y.
        • Paredes S.
        • Villanova L.
        • Brown K.
        • et al.
        SIRT7 represses Myc activity to suppress ER stress and prevent fatty liver disease.
        Cell Rep. 2013; 5: 654-665
        • Huang H.C.
        • Yu H.R.
        • Huang L.T.
        • Huang H.C.
        • Chen R.F.
        • Lin I.C.
        • et al.
        MiRNA-125b regulates TNF-α production in CD14+ neonatal monocytes via post-transcriptional regulation.
        J Leukoc Biol. 2012; 92: 171-182
        • Tili E.
        • Michaille J.J.
        • Cimino A.
        • Costinean S.
        • Dumitru C.D.
        • Adair B.
        • et al.
        Modulation of miR-155 and miR-125b levels following lipopolysaccharide/TNF-alpha stimulation and their possible roles in regulating the response to endotoxin shock.
        J Immunol. 2007; 179: 5082-5089
        • Scapa E.F.
        • Pocai A.
        • Wu M.K.
        • Gutierrez-Juarez R.
        • Glenz L.
        • Kanno K.
        • et al.
        Regulation of energy substrate utilization and hepatic insulin sensitivity by phosphatidylcholine transfer protein/StarD2.
        FASEB J. 2008; 22: 2579-2590
        • Matsuzaka T.
        • Shimano H.
        • Yahagi N.
        • Kato T.
        • Atsumi A.
        • Yamamoto T.
        • et al.
        Crucial role of a long-chain fatty acid elongase, Elovl6, in obesity-induced insulin resistance.
        Nat Med. 2007; 13: 1193-1202
        • Wang L.
        • Balas B.
        • Christ-Roberts C.Y.
        • Kim R.Y.
        • Ramos F.J.
        • Kikani C.K.
        • et al.
        Peripheral disruption of the Grb10 gene enhances insulin signaling and sensitivity in vivo.
        Mol Cell Biol. 2007; 27: 6497-6505