Abstract
This study was aimed to investigate the potential regulatory mechanism of high-content hydrogen water (HHW) in non-alcoholic fatty liver disease (NAFLD). A high-fat diet (HFD)-induced NAFLD mice model and cellular model were prepared. The serum levels of alanine transaminase (ALT), aspartate transaminase (AST), total cholesterol (TCH) and triglycerides (TG) were measured. The expression levels of representative five microRNA (miRNAs) (miR-103, miR-488, miR-136, miR-505 and miR-148a) in liver tissues were determined by quantitative real-time PCR (qRT-PCR). The target of miR-136 was validated by RNA immunoprecipitation (RIP) and pull-down assay. MiR-136, MEG3 and nuclear factor erythroid 2-related factor 2 (Nrf2) expression levels following cell treatment were detected in hepatocytes using qRT-PCR and Western blotting. Moreover, cell viability and TG content were conducted. MiR-136 was downregulated, MEG3 as well as Nrf2 was upregulated and serum lipid level was reduced in NAFLD mice model after HHW treatment, which exerted the same effect in cellular model. RIP and RNA pull-down assay confirmed that MEG2 was a downstream target of miR-136. What’s more, HHW ameliorated lipid accumulation by regulating miR-136/MEG3/Nrf2 axis in vitro and in vivo. Hence, HHW alleviated NAFLD by downregulation of miR-136 through mediating Nrf2 via targeting MEG3.
Acknowledgments
This study was supported by Natural Science Foundation of Henan Province (No. 162300410306), Funder Id: 10.13039/501100006407.
References
Al Zarzour, R.H., Ahmad, M., Asmawi, M.Z., Kaur, G., Maa, S., Almansoub, M.A., Sam, S., Usman, N.S., Aldulaimi, D.W., and Yam, M.F. (2017). Phyllanthus niruri standardized extract alleviates the progression of non-alcoholic fatty liver disease and decreases atherosclerotic risk in Sprague-Dawley Rats. Nutrients 9, 766.10.3390/nu9070766Search in Google Scholar PubMed PubMed Central
An, X., Yang, Z., and An, Z. (2017). MiR-149 compromises the reactions of liver cells to fatty acid via its polymorphism and increases non-alcoholic fatty liver disease (NAFLD) risk by targeting methylene tetrahydrofolate reductase (MTHFR). Med. Sci. Monit. 23, 2299–2307.10.12659/MSM.901377Search in Google Scholar
Anstee, Q.M., Targher, G., and Day, C.P. (2013). Progression of NAFLD to diabetes mellitus, cardiovascular disease or cirrhosis. Nat. Rev. Gastro. Hepat. 10, 330–344.10.1038/nrgastro.2013.41Search in Google Scholar PubMed
Bostjancic, E., Bandelj, E., Luzar, B., Poljak, M., and Glavac, D. (2015). Hepatic expression of miR-122, miR-126, miR-136 and miR-181a and their correlation to histopathological and clinical characteristics of patients with hepatitis C. J. Viral Hepat. 22, 146–157.10.1111/jvh.12266Search in Google Scholar PubMed
Chowdhry, S., Nazmy, M.H., Meakin, P.J., Dinkova-Kostova, A.T., Walsh, S.V., Tsujita, T., Dillon, J.F., Ashford, M.L.J., and Hayes, J.D. (2010). Loss of Nrf2 markedly exacerbates nonalcoholic steatohepatitis. Free Radic. Biol. Med. 48, 357–371.10.1016/j.freeradbiomed.2009.11.007Search in Google Scholar PubMed
Gupta, R.K., Patel, A.K., Shah, N., Chaudhary, A.K., Jha, U.K., Yadav, U.C., Gupta, P.K., and Pakuwal, U. (2014). Oxidative stress and antioxidants in disease and cancer: a review. Asian Pac. J. Cancer Prev. 15, 4405–4409.10.7314/APJCP.2014.15.11.4405Search in Google Scholar PubMed
Hahn, M.E., Timme-Laragy, A.R., Karchner, S.I., and Stegeman, J.J. (2015). Nrf2 and Nrf2-related proteins in development and developmental toxicity: insights from studies in zebrafish (Danio rerio). Free Radic. Biol. Med. 88, 275–289.10.1016/j.freeradbiomed.2015.06.022Search in Google Scholar PubMed PubMed Central
He, Y., Wu, Y.T., Huang, C., Meng, X.M., Ma, T.T., Wu, B.M., Xu, F.Y., Zhang, L., Lv, X.W., and Li, J. (2014). Inhibitory effects of long noncoding RNA MEG3 on hepatic stellate cells activation and liver fibrogenesis. Biochim. Biophys. Acta 1842, 2204–2215.10.1016/j.bbadis.2014.08.015Search in Google Scholar PubMed
Kawai, D., Takaki, A., Nakatsuka, A., Wada, J., Tamaki, N., Yasunaka, T., Koike, K., Tsuzaki, R., Matsumoto, K. and Miyake, Y. (2012). Hydrogen-rich water prevents progression of nonalcoholic steatohepatitis and accompanying hepatocarcinogenesis in mice. Hepatology 56, 912–921.10.1002/hep.25782Search in Google Scholar PubMed
Okamoto, K., Koda, M., Okamoto, T., Onoyama, T., Miyoshi, K., Kishina, M., Kato, J., Tokunaga, S., Sugihara, T.A., Hara, Y., et al. (2016). A series of microRNA in the chromosome 14q32.2 maternally imprinted region related to progression of non-alcoholic fatty liver disease in a mouse model. PLoS One 11, e0154676.10.1371/journal.pone.0154676Search in Google Scholar PubMed PubMed Central
Olaywi, M., Bhatia, T., Anand, S., and Singhal, S. (2013). Novel anti-diabetic agents in non-alcoholic fatty liver disease: a mini-review. Hepatobiliary Pancreat. Dis. Int. 12, 584–588.10.1016/S1499-3872(13)60092-2Search in Google Scholar PubMed
Runtuwene, J., Amitani, H., Amitani, M., Asakawa, A., Cheng, K.C., and Inui, A. (2015). Hydrogen-water enhances 5-fluorouracil-induced inhibition of colon cancer. Peer J. 3, e859.10.7717/peerj.859Search in Google Scholar PubMed PubMed Central
Tamaki, N., Orihuelacampos, R.C., Fukui, M., and Ito, H. (2016). Hydrogen-rich water intake accelerates oral palatal wound healing via activation of the Nrf2/antioxidant defense pathways in a rat model. Oxid. Med. Cell. Longev. 2016, 5679040.10.1155/2016/5679040Search in Google Scholar PubMed PubMed Central
Vos, M.B. (2014). Nutrition, nonalcoholic fatty liver disease and the microbiome: recent progress in the field. Curr. Opin. Lipidol. 25, 61–66.10.1097/MOL.0000000000000043Search in Google Scholar PubMed PubMed Central
Wang, F., Yu, G., Liu, S.Y., Li, J.B., Wang, J.F., Bo, L.L., Qian, L.R., Sun, X.J., and Deng, X.M. (2011). Hydrogen-rich saline protects against renal ischemia/reperfusion injury in rats. J. Surg. Res. 167, e339.10.1016/j.jss.2010.11.005Search in Google Scholar PubMed
Wang, Y., Wang, J., Wei, L.J., Zhu, D.M., and Zhang, J.S. (2017). Biological function and mechanism of lncRNA-MEG3 in Tenon’s capsule fibroblasts proliferation: by MEG3-Nrf2 protein interaction. Biomed. Pharmacother. 87, 548–554.10.1016/j.biopha.2016.12.040Search in Google Scholar PubMed
Wong, R.J., Cheung, R., and Ahmed, A. (2014). Nonalcoholic steatohepatitis is the most rapidly growing indication for liver transplantation in patients with hepatocellular carcinoma in the U.S. Hepatology 59, 2188–2195.10.1002/hep.26986Search in Google Scholar PubMed
Yang, X. and Deng, F. (2017). Iron overload is associated with non-alcoholic fatty liver disease (NAFLD): results from The NHANES III survey. Eur. J. BioMed. Res. 3, 10–15.10.18088/ejbmr.3.1.2017.pp10-15Search in Google Scholar
Younossi, Z.M., Koenig, A.B., Abdelatif, D., Fazel, Y., Henry, L., and Wymer, M. (2016). Global epidemiology of nonalcoholic fatty liver disease – meta-analytic assessment of prevalence, incidence, and outcomes. Hepatology 64, 73–84.10.1002/hep.28431Search in Google Scholar PubMed
Zhou, Y., Zhang, X., and Klibanski, A. (2012). MEG3 noncoding RNA: a tumor suppressor. J. Mol. Endocrinol. 48, R45–R53.10.1530/JME-12-0008Search in Google Scholar PubMed PubMed Central
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