Advertisement

Rosiglitazone Inhibits Activation of Hepatic Stellate Cells via Up-Regulating Micro-RNA-124-3p to Alleviate Hepatic Fibrosis

  • Shao-ce Zhi
  • Shi-zuan Chen
  • Yan-yan Li
  • Jun-jian Li
  • Yi-hu ZhengEmail author
  • Fu-xiang YuEmail author
Original Article
  • 13 Downloads

Abstract

Background

The activation of hepatic stellate cells (HSCs) is involved in hepatic fibrogenesis and is regulated by the decreased expression of peroxisome proliferator-activated receptor γ (PPARγ). Rosiglitazone (RGZ) is a highly potent agonist of PPARγ.

Aims

To clarify molecular regulatory mechanism of RGZ in the activation of HSCs in hepatic fibrosis.

Methods

A mouse model of hepatic fibrosis was established by carbon tetrachloride with or without RGZ intervention. A vector carrying pcDNA-HOTAIR was constructed and injected into a mouse model. HSCs were isolated from liver tissue and activated by transforming growth factor-β. The expression of miR-124-3p, HOTAIR, Col1A1, α-SMA, and PPARγ mRNAs was measured by quantitative real-time PCR. The level of PPARγ was measured by Western blotting. The interaction between HOTAIR and PPARγ was assessed by RNA immunoprecipitation (RIP) and RNA pull-down. The target gene of miR-124-3p was determined by luciferase reporter assay and RNA interference approaches.

Results

The expression of Col1A1 and α-SMA was reduced after RGZ intervention. Different expressions of HOTAIR and miR-124-3p were observed in liver tissue and HSCs. The luciferase reporter assay and RNA interference approaches indicated that miR-124-3p negatively regulated HOTAIR expression. RIP and RNA pull-down results revealed that PPARγ was interacted by HOTAIR. The therapeutic effect of RGZ on hepatic fibrosis was reversed by overexpression of HOTAIR.

Conclusions

RGZ inhibits the activation of HSCs by up-regulating miR-124-3p. The silencing of HOTAIR by miR-124-3p in HSC activation provided the foundation to understand interactions of ncRNAs and potential treatment target in hepatic fibrosis.

Keywords

Hepatic fibrosis HOTAIR MiR-124-3p PPARγ Rosiglitazone 

Notes

Acknowledgments

This study was funded by Zhejiang Provincial Natural Science Foundation (Grant Number: LY16H030014) and Zhejiang Provincial Medical Technology Project (Grant Number: 2019KY104) and Zhejiang Provincial Science Technology Project (Grant Number: 2015C37101).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

This study was approved by the Institutional Ethics Committee of the First Affiliated Hospital of Wenzhou Medical University. All of these experiments in the current research were in compliance with the government policies and defined protocols.

Informed consent

Informed consent was obtained from all individual participants included in the study.

Availability of data and material

The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.

References

  1. 1.
    Trautwein C, Friedman SL, Schuppan D, Pinzani M. Hepatic fibrosis: concept to treatment. J Hepatol. 2015;62:S15–S24.CrossRefGoogle Scholar
  2. 2.
    Mehal WZ, Schuppan D. Antifibrotic therapies in the liver. Semin Liver Dis. 2015;35:184–198.CrossRefGoogle Scholar
  3. 3.
    Hsu W-H, Lee BH, Hsu YW, Pan TM. Peroxisome proliferator-activated receptor-gamma activators monascin and rosiglitazone attenuate carboxymethyllysine-induced fibrosis in hepatic stellate cells through regulating the oxidative stress pathway but independent of the receptor for advanced glycation end products signaling. J Agric Food Chem. 2013;61:6873–6879.CrossRefGoogle Scholar
  4. 4.
    Kersten S, Desvergne B, Wahli W. Roles of PPARs in health and disease. Nature. 2000;405:421.CrossRefGoogle Scholar
  5. 5.
    Marra F, Efsen E, Romanelli RG, et al. Ligands of peroxisome proliferator-activated receptor gamma modulate profibrogenic and proinflammatory actions in hepatic stellate cells. Gastroenterology. 2000;119:466–478.CrossRefGoogle Scholar
  6. 6.
    Miyahara T, Schrum L, Rippe R, et al. Peroxisome proliferator-activated receptors and hepatic stellate cell activation. J Biol Chem. 2000;275:35715–35722.CrossRefGoogle Scholar
  7. 7.
    Lu Y, Ma D, Xu W, Shao S, Yu X. Effect and cardiovascular safety of adding rosiglitazone to insulin therapy in type 2 diabetes: a meta-analysis. J Diabetes Investig. 2015;6:78–86.CrossRefGoogle Scholar
  8. 8.
    Zhao C, Chen W, Yang L, Chen L, Stimpson SA, Diehl AM. PPARgamma agonists prevent TGFbeta1/Smad3-signaling in human hepatic stellate cells. Biochem Biophys Res Commun. 2006;350:385–391.CrossRefGoogle Scholar
  9. 9.
    Asano T, Yamazaki H, Kasahara C, et al. Identification, synthesis, and biological evaluation of 6-[(6R)-2-(4-fluorophenyl)-6-(hydroxymethyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyr imidin-3-yl]-2-(2-methylphenyl)pyridazin-3(2H)-one (AS1940477), a potent p38 MAP kinase inhibitor. J Med Chem. 2012;55:7772–7785.CrossRefGoogle Scholar
  10. 10.
    Yoshihara D, Kurahashi H, Morita M, et al. PPARgamma agonist ameliorates kidney and liver disease in an orthologous rat model of human autosomal recessive polycystic kidney disease. Am J Physiol Renal Physiol. 2011;300:F465–F474.CrossRefGoogle Scholar
  11. 11.
    Cech TR, Steitz JA. The noncoding RNA revolution-trashing old rules to forge new ones. Cell. 2014;157:77–94.CrossRefGoogle Scholar
  12. 12.
    Zell S, Schmitt R, Witting S, Kreipe HH, Hussein K, Becker JU. Hypoxia induces mesenchymal gene expression in renal tubular epithelial cells: an in vitro model of kidney transplant fibrosis. Nephron Extra. 2013;3:50–58.CrossRefGoogle Scholar
  13. 13.
    Gong M, Liang T, Jin S, et al. Methylation-mediated silencing of miR-124 facilitates chondrogenesis by targeting NFATc1 under hypoxic conditions. Am J Transl Res. 2017;9:4111–4124.PubMedPubMedCentralGoogle Scholar
  14. 14.
    Wang D, Zhang H, Li M, et al. MicroRNA-124 controls the proliferative, migratory, and inflammatory phenotype of pulmonary vascular fibroblasts. Circ Res. 2014;114:67–78.CrossRefGoogle Scholar
  15. 15.
    Jiang J, Gusev Y, Aderca I, et al. Association of microRNA expression in hepatocellular carcinomas with hepatitis infection, cirrhosis, and patient survival. Clin Cancer Res. 2008;14:419–427.CrossRefGoogle Scholar
  16. 16.
    Jiang X-P, Ai WB, Wan LY, Zhang YQ, Wu JF. The roles of microRNA families in hepatic fibrosis. Cell Biosci. 2017;7:34.CrossRefGoogle Scholar
  17. 17.
    Wang Y, Hu M. Peroxisome proliferators activated receptor γ protects against acute lung injury alveolar macrophages inflammation by upregulating miR-124 expression. Chin J Lung Dis. 2015;8:160–165.Google Scholar
  18. 18.
    Carninci P, Kasukawa T, Katayama S, et al. The transcriptional landscape of the mammalian genome. SPJ. 2005;309:1559–1563.Google Scholar
  19. 19.
    Sun M, Kraus WL. From discovery to function: the expanding roles of long noncoding RNAs in physiology and disease. Endocr Rev. 2015;36:25–64.CrossRefGoogle Scholar
  20. 20.
    Bian EB, Wang YY, Yang Y, et al. Hotair facilitates hepatic stellate cells activation and fibrogenesis in the liver. Biochimica et Biophysica Acta. 2017;1863:674–686.CrossRefGoogle Scholar
  21. 21.
    Rinn JL, Kertesz M, Wang JK, et al. Functional demarcation of active and silent chromatin domains in human HOX loci by non-coding RNAs. Cell. 2007;129:1311–1323.CrossRefGoogle Scholar
  22. 22.
    Chiyomaru T, Fukuhara S, Saini S, et al. Long non-coding RNA HOTAIR is targeted and regulated by miR-141 in human cancer cells. J Biol Chem. 2014;289:12550–12565.CrossRefGoogle Scholar
  23. 23.
    Bennett RG, Simpson RL, Hamel FG. Serelaxin increases the antifibrotic action of rosiglitazone in a model of hepatic fibrosis. World J Gastroenterol. 2017;23:3999–4006.CrossRefGoogle Scholar
  24. 24.
    Zhang K, Han X, Zhang Z, et al. The liver-enriched lnc-LFAR1 promotes liver fibrosis by activating TGFβ and Notch pathways. Nat Commun. 2017;8:144.CrossRefGoogle Scholar
  25. 25.
    Ikeda K, Wakahara T, Wang YQ, Kadoya H, Kawada N, Kaneda K. In vitro migratory potential of rat quiescent hepatic stellate cells and its augmentation by cell activation. Hepatology. 1999;29:1760–1767.CrossRefGoogle Scholar
  26. 26.
    Friedman SL. Molecular regulation of hepatic fibrosis, an integrated cellular response to tissue injury. J Biol Chem. 2000;28:2247–2250.CrossRefGoogle Scholar
  27. 27.
    Armoni M, Harel C, Karnieli E. PPARγ gene expression is autoregulated in primary adipocytes: ligand, sumoylation, and isoform specificity. Horm Metab Res. 2015;47:89–96.PubMedGoogle Scholar
  28. 28.
    Zhang Q, Xiang S, Liu Q, et al. PPARγ antagonizes hypoxia-induced activation of hepatic stellate cell through cross mediating PI3K/AKT and cGMP/PKG signaling. PPAR Res. 2018.  https://doi.org/10.1155/2018/6970407.CrossRefPubMedPubMedCentralGoogle Scholar
  29. 29.
    Kweon S-M, Chi F, Higashiyama R, Lai K, Tsukamoto H. Wnt pathway stabilizes MeCP2 protein to repress PPAR-γ in activation of hepatic stellate cells. PLoS ONE. 2016;11:e0156111.CrossRefGoogle Scholar
  30. 30.
    Troeger JS, Mederacke I, Gwak GY, et al. Deactivation of hepatic stellate cells during liver fibrosis resolution in mice. Gastroenterology. 2012;143:1073.  https://doi.org/10.1053/j.gastro.2012.06.036.CrossRefPubMedGoogle Scholar
  31. 31.
    Morán-Salvador E, Titos E, Rius B, et al. Cell-specific PPARγ deficiency establishes anti-inflammatory and anti-fibrogenic properties for this nuclear receptor in non-parenchymal liver cells. J Hepatol. 2013;59:1045–1053.CrossRefGoogle Scholar
  32. 32.
    Odegaard JI, Ricardo-Gonzalez RR, Eagle AR, et al. Alternative M2 activation of Kupffer cells by PPARδ ameliorates obesity-induced insulin resistance. Cell Metab. 2008;7:496–507.CrossRefGoogle Scholar
  33. 33.
    Neuschwander-Tetri BA, Loomba R, Sanyal AJ, et al. Farnesoid X nuclear receptor ligand obeticholic acid for non-cirrhotic, nonalcoholic steatohepatitis (FLINT): a multicentre, randomised, placebo-controlled trial. Lancet. 2015;385:956–965.CrossRefGoogle Scholar
  34. 34.
    Yang J-J, Tao H, Li J. Hedgehog signaling pathway as key player in liver fibrosis: new insights and perspectives. Expert Opin Therap Targets. 2014;18:1011–1021.CrossRefGoogle Scholar
  35. 35.
    Noetel A, Kwiecinski M, Elfimova N, Huang J, Odenthal M. microRNA are central players in anti- and profibrotic gene regulation during liver fibrosis. Front Physiol. 2012;3:49.CrossRefGoogle Scholar
  36. 36.
    Roderburg C, Urban GW, Bettermann K, et al. Micro-RNA profiling reveals a role for miR-29 in human and murine liver fibrosis. Hepatology. 2010;53:209–218.CrossRefGoogle Scholar
  37. 37.
    Lakner AM, Steuerwald NM, Walling TL, et al. Inhibitory effects of microRNA 19b in hepatic stellate cell-mediated fibrogenesis. Hepatology. 2012;56:300–310.CrossRefGoogle Scholar
  38. 38.
    Liang YG, Liu ZT, Guo SX. Ultrasound reverses adriamycin-resistance in non-small cell lung cancer via positive regulation of BRAF-activated non-coding RNA (BANCR) expression. Clin Surg Res Commun. 2017;1:18–23.CrossRefGoogle Scholar
  39. 39.
    Zhang A, Zhao JC, Kim J, et al. LncRNA HOTAIR enhances the androgen-receptor-mediated transcriptional program and drives castration-resistant prostate cancer. Cell Rep. 2015;13:209–221.CrossRefGoogle Scholar
  40. 40.
    Bhan A, Mandal SS. LncRNA HOTAIR: a master regulator of chromatin dynamics and cancer. Biochim Biophys Acta. 2015;1856:151–164.PubMedPubMedCentralGoogle Scholar
  41. 41.
    Shenoy A, Blelloch RH. Regulation of microRNA function in somatic stem cell proliferation and differentiation. Nat Rev Mol Cell Biol. 2014;15:565–576.CrossRefGoogle Scholar
  42. 42.
    Song R, Walentek P, Sponer N, et al. miR-34/449 miRNAs are required for motile ciliogenesis by repressing cp110. Nature. 2014;510:115–120.CrossRefGoogle Scholar
  43. 43.
    Sun Q, Csorba T, Skourti-Stathaki K, Proudfoot NJ, Dean C. R-loop stabilization represses antisense transcription at the Arabidopsis FLC locus. Science. 2013;340:619–621.CrossRefGoogle Scholar
  44. 44.
    Ghosal S, Das S, Sen R, Basak P, Chakrabarti J. Circ2Traits: a comprehensive database for circular RNA potentially associated with disease and traits. Front Genet. 2013;4:283.CrossRefGoogle Scholar
  45. 45.
    Wang J, Liu X, Wu H, et al. CREB up-regulates long non-coding RNA, HULC expression through interaction with microRNA-372 in liver cancer. Nucl Acids Res. 2010;38:5366–5383.CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of Hepatobiliary and Pancreatic Surgery, The First Affiliated HospitalWenzhou Medical UniversityWenzhouPeople’s Republic of China

Personalised recommendations