The effect of microRNA-219 on lipid metabolism, cell viability and apoptosis of hepatocytes under lipotoxic environment

Abstract

Abstract: Objective To explore the effect of microRNA-219 (miR-219) on lipid metabolism, cell viability and apoptosis of lipotoxic hepatocytes.Methods Mice were fed with low-fat diet (LFD) and high-fat diet (HFD) to construct control group and non-alcoholic fatty liver diseases (NAFLD) model. Hematoxylin-eosin (H＆E) staining and oil red staining were used to observe the liver histology. Primary hepatocytes (PHCs) were isolated from wild-type mice and then incubated with 200uM palmitic acid (PA) for 24h to mimic a high-fat environment. Fatty degeneration of hepatocytes was judged by oil red staining, aspartate transaminase (AST) and alanine aminotransferase (ALT) levels. Quantitative polymerase chain reaction (q-PCR) was used to detect the expression of miR-219 in liver tissue and PHCs, and the mRNA expression of lipid metabolism factors [stearoyl-CoA desaturase 1 (SCD1) and fatty acid synthase (FASN)], proliferation-related factors [proliferating cell nuclear antigen (PCNA) and CyclinD1] and apoptosis-related factors [B-cell lymphoma-2 (BCL-2) and Caspase3] after overexpression of miR-219 through miR-219 mimic (mi-miR-219) or the negative control in PA-treated PHCs. Oil red staining was performed to observe cell lipid deposition. EDU was used to observe cell proliferation.Results The NAFLD mice model was successfully established. Compared with the relatively control group, miR-219 expression was significantly decreased in both the liver tissues of HFD group and PA-treated PHCs [HFD group: (0.455±0.028), LFD group: (1.000±0.125), P＜0.05; PA-treated group: (0.676±0.064), Control: (1.000±0.190), P＜0.05]. After overexpressing miR-219, SCD1 and FASN mRNA expression were both significantly decreased in PA-treated PHCs [SCD1: mi-miR-219+PA: (0.539±0.048), negative control group (miR-NC)+PA: (1.000±0.033), P＜0.001; FASN: mi-miR-219+PA: (0.722±0.036), miR-NC+PA: (1.000±0.051), P＜0.05]. Oil red staining indicated that the lipid deposition was also significantly alleviated in PA-treated PHC in mi-miR-219 group. Compared with the miR-NC group, the mRNA expression of proliferation-related factors was both obviously increased [PCNA: mi-miR-219+PA: (1.652±0.185), miR-NC+PA: (1.000±0.203), P＜0.05; CyclinD1: mi-miR-219+PA: (1.791±0.154), miR-NC+PA: (1.000±0.135), P＜0.05]. And the more obvious red fluorescent was showed in mi-miR-219 PHCs group according to EDU assay. Besides, the mRNA expression of pro-apoptotic factor Caspase-3 was decreased [mi-miR-219+PA: (0.574±0.054), miR-NC+PA: (1.000±0.35), P＜0.05], but the mRNA expression of anti-apoptotic factor BCL-2 was increased [mi-miR-291: (1.535±0.109), miR-NC: (1.000±0.208), P＜0.05].Conclusion Under high-fat environment, miR-219 could significantly alleviate hepatocytes lipid deposition, promote hepatocytes viability and inhibit cell apoptosis to delay the progress of NAFLD.

Key words: MicroRNA-219, Lipid metabolism, Cell viability and apoptosis, Non-alcoholic fatty liver diseases

XU Zi-xin, LIAO Hong-yu, LI Hui-yi, XU Ming-yi. The effect of microRNA-219 on lipid metabolism, cell viability and apoptosis of hepatocytes under lipotoxic environment[J]. Chinese Hepatolgy, 2022, 27(8): 920-924.

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