Resveratrol ameliorates nutritional steatohepatitis through the mmu‑miR‑599/PXR pathway.

Abstract

The aim of the present study was to elucidate the effect of resveratrol on non-alcoholic steatohepatitis (NASH), and the molecular basis in mice and Hepa1-6 cells, in order to verify its therapeutic effect. C57BL/6J mice were fed a methionine-choline-deficient (MCD) diet to induce steatohepatitis and were treated with resveratrol. Mouse sera were collected for biochemical analysis and enzyme-linked immunosorbent assay, and livers were obtained for histological observation, and mmu-microRNA (miR)-599 and inflammation-related gene expression analysis. Hepa1-6 cells were treated with palmitic acid to establish a NASH cell model, and were then treated with resveratrol, or transfected with mmu-miR-599 mimic, mmu-miR-599 inhibitor or recombinant pregnane X receptor (PXR) plasmid. Subsequently, the cells were collected for mmu-miR-599 and inflammation-related gene expression analysis. Reverse transcription-quantitative polymerase chain reaction and western blotting were used to assess mmu-miR-599 expression levels, and the mRNA and protein expression levels of PXR and inflammation-related genes. The binding site of mmu-miR-599 in the PXR mRNA was verified by the luciferase activity assay. Mice fed an MCD diet for 4 weeks exhibited steatosis, focal necrosis and inflammatory infiltration in the liver. Resveratrol significantly reduced serum aminotransferase and malondialdehyde levels, and ameliorated hepatic injury. These effects were associated with reduced mmu-miR-599 expression, enhanced PXR expression, and downregulated levels of nuclear factor-κB, tumour necrosis factor-α, interleukin (IL)-1β, IL-6, NOD-like receptor family pyrin domain-containing protein 3 and signal transducer and activator of transcription 3. Administration of the mmu-miR-599 mimic inhibited PXR expression in Hepa1-6 cells, whereas the mmu-miR-599 inhibitor exerted the opposite effect. A binding site for mmu-miR-599 was identified in the PXR mRNA sequence. Furthermore, overexpression of PXR inhibited the expression of inflammatory factors in Hepa1-6 cells. The present study provided evidence for the protective role of resveratrol in ameliorating steatohepatitis through regulating the mmu-miR-599/PXR pathway and the consequent suppression of related inflammatory factors. Resveratrol may serve as a potential candidate for steatohepatitis management.