RESEARCH COMMUNICATION Inhibiting miR-27a and miR-142-5p attenuate nonalcoholic fatty liver disease by regulating Nrf2 signaling pathway Maryam Teimouri 1 | Hossein Hosseini 1 | Maryam Shabani 1 | Mehdi Koushki 1 | Farshid Noorbakhsh 2 | Reza Meshkani 1 1 Department of Clinical Biochemistry, Faculty of Medicine, Tehran University of Medical Sciences, Tehran, Iran 2 Department of Immunology, Faculty of Medicine, Tehran University of Medical Sciences, Tehran, Iran Correspondence Reza Meshkani, Department of Clinical Biochemistry, Faculty of Medicine, Tehran University of Medical Sciences, Tehran, Iran. Email: rmeshkani@tums.ac.ir Funding information Tehran University of Medical Sciences and Health Services, Grant/Award Number: grant 96-01-30-33971 Summary The gene Nrf2 (nuclear factor-erythroid 2-related factor 2) is the most important regulator of the cellular antioxidant system and its dysregulation has a role in the etiology of nonalcoholic fatty liver disease (NAFLD). The aim of this study was to investigate the association between Nrf2 targeted miRNAs (miR-27a, miR-142-5p, miR-153, and miR-128) with lipid accumulation in vitro and in vivo models of NAFLD. We used two in vivo and in vitro models of NAFLD. The expression of the genes and miRNAs was assessed by real-time PCR and the protein level was evaluated using western blot. To investigate the potential role of miRNAs in NAFLD, the inhibitors or mimics of the miR-27a and miR-142-5p were trans- fected into HepG2 cells. The mRNA and protein levels of Nrf2 were significantly decreased in the liver of high fat diet-fed mice as well as in HepG2 cells treated with high glucose (HG). Reduced expression of Nrf2 was associated with increased expression levels of miR-27a and miR-142-5p in both models of NAFLD. HG-induced triglyceride accumulation was attenuated by inhibition of miR-27a or miR-142-5p in HepG2 cells. Overexpression of miR-27a or miR-142-5p suppressed the expression of Nrf2 and its downstream antioxidant genes and increased production of reactive oxygen species, whereas inhibition of miR-27a or miR-142-5p reversed these effects. In conclusion, the data of this study may sug- gest that miR-27a and miR-142-5p are increased in NAFLD, where they suppress Nrf2 expression and contribute to the accumulation of lipids in the hepatocytes. KEYWORDS lipogenesis, miR-27a, miR-142-5p, NAFLD, Nrf2, oxidative stress Abbreviations: ARE, antioxidant responsive elements; BSA, bovine serum albumin; DCFH-DA, dichlorodihydro-fluorescindiacetate; DMEM, dulbecco's modified Eagle's medium; ECL, enhanced chemiluminescence; EDTA, ethylenediaminetetraacetic acid; FAS, fatty acid synthase; FBS, fasting blood sugar; HG, high glucose; HFD, high fat diet; HO1, heme oxygenase 1; Keap1, kelch-like ECH-associated protein 1; Mn-SOD, manganese superoxide dismutase; NAFLD, nonalcoholic fatty liver disease; NC, negative control; NG, normal glucose; NQO1, NAD(P)H:Quinone Oxidoreductase 1; Nrf2, nuclear factor erythroid 2-related factor 2; PMSF, phenylmethylsulfonyl fluoride; PVDF, polyvinylidene difluoride; ROS, reactive oxygen species; RIPA, radioimmuno-precipitation; SCD, standard chow diet; SDS-PAGE, sodium dodecyl sulfate-polyacrylamide gel electrophoresis; SREBP- 1c, Sterol regulatory element-binding transcription factor 1c; TG, triglyceride. Received: 30 November 2019 Accepted: 16 December 2019 DOI: 10.1002/iub.2221 © 2019 International Union of Biochemistry and Molecular Biology IUBMB Life. 2020;72:361–372. wileyonlinelibrary.com/journal/iub 361