miR-125b-5p regulates FFA-induced hepatic steatosis in L02 cells by targeting estrogen-related receptor alpha

IF 2.6 3区生物学 Q2 GENETICS & HEREDITY

Gene Pub Date : 2025-03-18 DOI:10.1016/j.gene.2025.149419

Fen Gao , Yanhua Ma , Chun Yu , Qianchen Duan

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引用次数: 0

Abstract

Background & Aims

NAFLD is a global and complex liver disease caused by multiple factors. Intrahepatocellular steatosis is the primary prerequisite for the occurrence and development of NAFLD. It has been shown that miR-125b-5p is highly correlated with NAFLD, and ESRRA is a factor that regulates lipid metabolism. The purpose of our study is to investigate whether miR-125b-5p regulates FFA-induced steatosis in L02 cells by targeting ESRRA.

Approaches and results

Estrogen-related receptor alpha (ESRRA) was identified as a direct target of miR-125b-5p through database prediction and a dual-luciferase reporter gene assay. L02 cells were induced with free fatty acids (OA:PA, 2:1) at concentrations of 0.3 mM, 0.6 mM, 0.9 mM, 1.2 mM and 1.5 mM for 24 h, 48 h and 72 h, respectively. The degree of hepatocyte steatosis and triglyceride content were separately manifested by oil red O staining and colorimetric method. Cell viability per group was detected by CCK-8 assay. Eventually, 0.9 mM and 24 h were screened out as the optimal concentration and time for establishing the in-vitro model of hepatic steatosis. Followingly, miR-125b-5p and ESRRA were knocked down by transient transfection. We monitored the expressions of lipid metabolism factors SREBP-1c, ACC1 and FAS and determine triglyceride content within the cells per group. The data showed that knockdown of ESRRA led to down-regulation of the expressions of SREBP-1, ACC1, FAS and triglyceride content. Meanwhile, knockdown of ESRRA and miR-125b-5p resulted that the expressions of ESRRA, SREBP-1, ACC1, FAS and triglyceride content rebounded.

Conclusions

MiR-125b-5p down-regulates the expressions of lipid metabolism-related factors by negatively regulating ESRRA, thereby improving hepatic steatosis.

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miR-125b-5p 通过靶向雌激素相关受体 alpha 调节脂肪酸诱导的 L02 细胞肝脂肪变性。

背景与目的：非酒精性脂肪肝是由多种因素引起的全球性复杂肝病。肝细胞内脂肪变性是非酒精性脂肪肝发生和发展的首要前提。有研究表明，miR-125b-5p 与 NAFLD 高度相关，而 ESRRA 是调节脂质代谢的一个因子。我们的研究旨在探讨miR-125b-5p是否通过靶向ESRRA来调节FFA诱导的L02细胞脂肪变性：通过数据库预测和双荧光素酶报告基因检测，确定雌激素相关受体α（ESRRA）为miR-125b-5p的直接靶标。分别以 0.3 mM、0.6 mM、0.9 mM、1.2 mM 和 1.5 mM 的浓度诱导 L02 细胞 24 小时、48 小时和 72 小时。肝细胞脂肪变性程度和甘油三酯含量分别用油红 O 染色法和比色法表示。用 CCK-8 法检测各组细胞的活力。最终筛选出 0.9 mM 和 24 h 是建立肝脂肪变性体外模型的最佳浓度和时间。随后，通过瞬时转染敲除了 miR-125b-5p 和 ESRRA。我们监测了脂质代谢因子 SREBP-1c、ACC1 和 FAS 的表达。数据显示，敲除 ESRRA 会导致 SREBP-1、ACC1 和 FAS 的表达下调。同时，敲除 ESRRA 和 miR-125b-5p 会导致 ESRRA、SREBP-1、ACC1 和 FAS 的表达回升：结论：miR-125b-5p 可通过负调控 ESRRA 来下调脂质代谢相关因子的表达，从而改善肝脂肪变性。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Gene 生物-遗传学

CiteScore

6.10

自引率

2.90%

发文量

718

审稿时长

42 days

期刊介绍： Gene publishes papers that focus on the regulation, expression, function and evolution of genes in all biological contexts, including all prokaryotic and eukaryotic organisms, as well as viruses.