饱和脂肪酸诱导肝细胞系建立代谢相关脂肪性肝病细胞损伤模型

摘要 目的：探讨不同脂肪酸对肝细胞系脂质积累、细胞损伤的影响，选择合适诱导试剂及肝细胞系建立一种具有严重细胞损伤及炎症反应的晚期代谢相关脂肪性肝病（MAFLD）体外细胞模型。方法：以油酸（OA）或棕榈酸（PA）或其混合物分别处理HepG2和LO2细胞，以CCK8检测细胞存活率；以油红O染色及甘油三酯酶法检测细胞脂质积累程度；以qRT-PCR检测凋亡相关蛋白、纤维化相关蛋白、自噬相关蛋白、炎症因子的mRNA表达水平。结果：0.25 mmol/LPA作用HepG2细胞24 h可显著诱导甘油三酯（TG）和脂质积累，但对LO2细胞无明显影响；0.25 mmol/L PA处理两种细胞系可诱导显著的细胞损伤及炎症，OA可缓解PA对细胞的损伤作用。结论：利用PA处理HepG2细胞可引起一定程度的脂质积累，诱导显著的细胞损伤及炎症，是合适的MAFLD体外细胞模型。

Saturated Fatty Acids Induced Liver Cell Lines to Establish a Cell Injury Model of MAFLD in Vitro

ABSTRACT Objective: To investigate the effects of different fatty acids on lipid accumulation and cell injury in liver cell lines, and to choose a suitable reagent and liver cell lines to establish a cell model of advanced metabolic dysfunction-associated fatty liver disease (MAFLD) with severe cell damage and inflammatory response. Methods: HepG2 and LO2 cells were exposed to oleic acid (OA) or palmitic acid (PA) or their combination at ratios of 2:1 respectively. Cell Counting Kit-8 (CCK-8) assay was carried out to assess cell proliferation; the degree of lipid accumulation was detected by oil red O staining and triglyceride enzyme assay; the mRNA expression levels of apoptosis-related proteins, fibrosis-related proteins, autophagy-related proteins, and inflammatory factors were detected by qRT-PCR. Results: HepG2 cells were treated with 0.25 mmol/L PA for 24 h, which significantly induced the accumulation of TG and lipids, but had no significant effect on LO2 cells. Two cells were treated with OA alone or a mixture of OA and PA-induced lower cytotoxicity with less weakened functional capacity than did PA alone. Conclusion: HepG2 cells were treated with PA alone is more suitable for establishing a kind of serious liver cell injury and inflammation of MAFLD cell model.