乙酸钠对油酸诱导的BRL-3A细胞脂肪变性的影响及机制分析

doi:10.11843/j.issn.0366-6964.2022.12.030

摘要/Abstract

摘要： 通过向油酸诱导的大鼠肝成纤维细胞(BRL-3A)脂肪变性模型中添加不同浓度(2、4、8 mmol·L^-1)的乙酸钠，探讨其对脂肪变性细胞模型脂代谢的调控机理及细胞损伤的修复作用。试验方法：1)用不同浓度的油酸(0、0.03、0.06、0.12、0.24、0.48) mmol·L^-1刺激BRL-3A细胞24 h后，分别检测细胞相对活力、总脂滴面积、三酰甘油(TG)含量、天门冬氨酸氨基转移酶(AST)及丙氨酸氨基转移酶(ALT)活性，建立脂肪变性细胞模型；2)向BRL-3A细胞中添加不同浓度的乙酸钠，通过流式细胞术检测细胞凋亡率；3)用不同浓度的乙酸钠和0.12 mmol·L^-1油酸共同孵育BRL-3A细胞，试验分为4组，分别为油酸处理组、2 mmol·L^-1乙酸钠+油酸处理组、4 mmol·L^-1乙酸钠+油酸处理组和8 mmol·L^-1乙酸钠+油酸处理组，分别对细胞脂滴、TG含量、AST、ALT活性、AMPK信号通路蛋白以及脂代谢关键基因进行检测。结果显示:1)用0.12 mmol·L^-1油酸处理BRL-3A细胞24 h，成功建立BRL-3A细胞脂肪变性模型。2)不同浓度的乙酸钠对BRL-3A细胞凋亡率没有影响；3)4、8 mmol·L^-1乙酸钠处理脂肪变性细胞模型后，与油酸处理组相比，细胞总脂滴面积、每平方毫米脂滴数、TG含量、AST和ALT活性均显著(P<0.05)或极显著(P<0.01)下降，P-AMPK表达水平显著(P<0.05)或极显著(P<0.01)上升；脂合成代谢相关基因ACC、FAS以及SCD-1 mRNA表达水平均有一定程度下降；脂分解代谢相关基因CPT-1、CPT-2以及ACO mRNA表达水平均有一定程度上升。本研究表明，乙酸钠会通过AMPK通路激活脂分解代谢，减轻肝细胞脂质蓄积，并且对油酸诱导的BRL-3A细胞脂肪变性模型的损伤具有一定缓解作用。

关键词: BRL-3A细胞, 乙酸钠, 脂肪变性细胞模型, 脂代谢

Abstract: Sodium acetate at different concentrations (2, 4, 8 mmol·L^-1) was added to oleic acid-induced steatosis model of normal rat liver cells (BRL-3A). To investigate the regulation mechanism of steatosis cells model lipid metabolism and repair of cell damage, the following experiments were conducted: 1) BRL-3A cells were stimulated with oleic acid at different concentrations (0, 0.03, 0.06, 0.12, 0.24, 0.48 mmol·L^-1) for 24 h. The cell relative activity, total lipid droplet area, triglyceride (TG) content, aspartate aminotransferase (AST) and alanine aminotransferase (ALT) activities were detected to establish cell steatosis model. 2) BRL-3A cells were added with different concentrations of sodium acetate, and the apoptosis rate was detected by flow cytometry. 3)BRL-3A cells were incubated with different concentrations of sodium acetate and 0.12 mmol·L^-1 oleic acid. The experiment was divided into 4 groups, including oleic acid treatment group, 2 mmol·L^-1 sodium acetate + oleic acid treatment group, 4 mmol·L^-1 sodium acetate + oleic acid treatment group and 8 mmol·L^-1 sodium acetate + oleic acid treatment group. Lipid droplets and TG content, AST and ALT activities, AMPK signaling pathway proteins and key genes of lipid metabolism were detected. Results: 1) BRL-3A cells were treated with 0.12 mmol·L^-1 oleic acid for 24 h, and the steatosis model of BRL-3A cells was established successfully. 2)Different concentrations of sodium acetate had no effect on the apoptosis rate of BRL-3A cells. 3)Compared with oleic acid treatment group, the total lipid drop area, lipid drop number per mm², TG content, AST and ALT activities of cell steatosis model treated with 8 mmol·L^-1 sodium acetate were significantly decreased (P<0.05) or extremely significantly decreased (P<0.01). The expression level of P-AMPK was significantly (P<0.05) or extremely significantly increased (P<0.01). The mRNA expression levels of lipid anabolism-related genes ACC, FAS and SCD-1 all decreased to a certain extent; the mRNA expression levels of lipid catabolism-related genes CPT-1, CPT-2 and ACO all increased to a certain extent. This study showed that sodium acetate activates lipid catabolism through the AMPK pathway, attenuated the lipid accumulation in hepatocytes, and alleviated the oleic acid-induced damage of BRL-3A cell steatosis model.

Key words: BRL-3A cells, sodium acetate, steatosis cell model, lipid metabolism

中图分类号:

S856.5

李林, 宫彬彬, 王广力, 赵梅, 张源淑. 乙酸钠对油酸诱导的BRL-3A细胞脂肪变性的影响及机制分析[J]. 畜牧兽医学报, 2022, 53(12): 4450-4460.

LI Lin, GONG Binbin, WANG Guangli, ZHAO Mei, ZHANG Yuanshu. Effect of Sodium Acetate on Oleic Acid-induced Steatosis in BRL-3A Cells[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(12): 4450-4460.

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