目的 探讨α-亚麻酸植物甾醇酯（PS-ALA）对酒精（EtOH）诱导的肝细胞脂肪变性和氧化应激的改善作用,并基于SIRT3进一步探索潜在的分子机制。方法 采用EtOH诱导HepG2细胞脂肪变性和氧化应激模型,给予PS-ALA干预,油红O染色观察细胞脂质沉积,DCFH-DA荧光探针检测活性氧（ROS）水平,免疫荧光染色检测 SIRT3 的表达水平,蛋白免疫印迹检测SIRT3、PPARα、CPT-1A、SOD2和CYP2E1 蛋白表达水平。结果 与对照组相比,EtOH组细胞内出现了大量的脂质沉积并产生大量的ROS;PS-ALA 干预后显著改善 EtOH 诱导的细胞内脂质沉积,并有效抑制ROS生成;进一步的研究显示,PS-ALA显著上调脂肪酸β氧化关键调控信号分子SIRT3、PPARα和CPT-1A蛋白表达水平,同时增加SOD2蛋白表达,降低CYP2E1表达。结论 PS-ALA 能够抑制EtOH诱导的HepG2细胞脂肪变性和氧化应激,其机制可能与上调SIRT3表达水平和促进脂肪酸β氧化有关。
Abstract
Objective To investigate the ameliorative effect of plant sterol ester of α-linolenic acid (PS-ALA) on steatosis and oxidative stress in HepG2 cells exposed to ethanol (EtOH), and the underlying mechanism associated with the regulation of SIRT3 expression. Methods HepG2 cells were treated with EtOH and intervened by PS-ALA. The lipid accumulation in the cells was evaluated by Oil Red O staining. The level of reactive oxygen species (ROS) was detected by DCFH-DA fluorescent probe. The expression of SIRT3 was assessed by immunofluorescence staining. The protein expressions of SIRT3, PPARα, CPT-1A, SOD2 and CYP2E1 were measured by Western blot. Results Compared with the control group, the cells in the EtOH group showed a significant increase in intracellular lipid accumulation and ROS production. PS-ALA intervention significantly decreased lipid accumulation and suppressed ROS production. The results of further molecular mechanism studies showed that PS-ALA treatment significantly increased the protein expressions of SIRT3, PPARα, and CPT-1A. Meanwhile, PS-ALA treatment significantly increased the protein expression of SOD2 and decreased the protein expression of CYP2E1. Conclusion PS-ALA can inhibit EtOH-induced steatosis and oxidative stress in HepG2 cells, and its molecular mechanism is related to promoting β-oxidation of fatty acid and inhibiting oxidative stress by up-regulating SIRT3 expression.
关键词
α-亚麻酸植物甾醇酯 /
脂肪变性 /
SIRT3 /
HepG2细胞
Key words
plant sterol ester of α-linolenic acid /
steatosis /
SIRT3 /
HepG2 cell
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基金
国家自然科学基金（No.81602856）; 山西省应用基础研究（No.202303021211124）
