Department of Traditional and Western Medical Hepatology, Third Hospital of Hebei Medical University, Shijiazhuang, Hebei 050051, P.R. China.
Int J Mol Med. 2022 Apr;49(4). doi: 10.3892/ijmm.2022.5102. Epub 2022 Feb 9.
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.
本研究旨在阐明白藜芦醇对非酒精性脂肪性肝炎(NASH)的作用及其在小鼠和 Hepa1-6 细胞中的分子基础,以验证其治疗效果。将 C57BL/6J 小鼠用蛋氨酸-胆碱缺乏(MCD)饮食喂养以诱导脂肪性肝炎,并给予白藜芦醇治疗。收集小鼠血清进行生化分析和酶联免疫吸附试验,收集肝脏进行组织学观察以及检测 mmu-微 RNA(miR)-599 和炎症相关基因的表达。用棕榈酸处理 Hepa1-6 细胞建立 NASH 细胞模型,然后用白藜芦醇、mmu-miR-599 模拟物、mmu-miR-599 抑制剂或重组孕烷 X 受体(PXR)质粒处理细胞。随后,收集细胞进行 mmu-miR-599 和炎症相关基因表达分析。采用逆转录定量聚合酶链反应和蛋白质印迹法检测 mmu-miR-599 的表达水平以及 PXR 和炎症相关基因的 mRNA 和蛋白表达水平。通过荧光素酶活性测定验证 mmu-miR-599 在 PXR mRNA 上的结合位点。用 MCD 饮食喂养 4 周的小鼠表现出肝脂肪变性、局灶性坏死和炎症浸润。白藜芦醇显著降低血清转氨酶和丙二醛水平,改善肝损伤。这些作用与 mmu-miR-599 表达降低、PXR 表达增强以及核因子-κB、肿瘤坏死因子-α、白细胞介素(IL)-1β、IL-6、NOD 样受体家族吡啶结构域含蛋白 3 和信号转导和转录激活因子 3 水平降低有关。给予 mmu-miR-599 模拟物抑制 Hepa1-6 细胞中的 PXR 表达,而 mmu-miR-599 抑制剂则产生相反的作用。在 PXR mRNA 序列中鉴定到 mmu-miR-599 的结合位点。此外,PXR 的过表达抑制了 Hepa1-6 细胞中炎症因子的表达。本研究为白藜芦醇通过调节 mmu-miR-599/PXR 通路和抑制相关炎症因子来改善脂肪性肝炎提供了证据。白藜芦醇可能是治疗脂肪性肝炎的潜在候选药物。