泽泻提取物改善游离脂肪酸诱导的HepG2脂肪变性细胞模型的脂质代谢及氧化应激异常

目的 研究泽泻提取物（Alisma orientale extract,AE）及其3种主要单体成分对游离脂肪酸（free fatty acid,FFA）诱导的HepG2脂肪变性细胞模型的脂质代谢及氧化应激异常的改善作用及机制。方法 利用FFA（油酸-棕榈酸2∶1）建立稳定的HepG2脂肪变性细胞模型，CCK-8法测定AE、泽泻醇A、泽泻醇A-23-乙酸酯及泽泻醇A-24-乙酸酯对HepG2细胞的安全剂量；在FFA诱导同时给予AE(25.00、12.50、6.25 mg/L)、泽泻醇A(50.0、25.0、12.5μmol/L)、泽泻醇A-23-乙酸酯（25.00、12.50、6.25μmol/L）、泽泻醇A-24-乙酸酯（50.0、25.0、12.5μmol/L）处理24 h，检测细胞内脂滴的形成情况，利用生化试剂盒测定各组细胞内三酰甘油（triglyceride,TG）、总胆固醇（total cholesterol,TC）水平，DCFH-DA检测药物对细胞内活性氧（reactive oxygen species,ROS）生成的影响，采用试剂盒检测细胞内氧化应激关键指标丙二醛（malo...

Alisma orientale extract alleviates lipid metabolism and oxidative stress abnormalities in free fatty acid-induced HepG2 steatosis cell model

Objective To explore the ameliorative effect and mechanisms of Alisma orientale extract (AE) and its three primary components on lipid metabolism abnormalities and oxidative stress in free fatty acids (FFA)-induced HepG2 steatosis cell model. Methods A HepG2 steatosis cell model was established using FFA [oleic acid-palmitic acid (2∶1)]. Safe dosages of AE, alisol A, alisol A-23-acetate, and alisol A-24-acetate for HepG2 cells were determined using CCK-8 kit. Simultaneously administering AE (25.00, 12.50, 6.25 mg/L), alisol A (50.0, 25.0, 12.5 μmol/L), alisol A-23-acetate (25.00, 12.50, 6.25 μmol/L), and alisol A-24-acetate (50.0, 25.0, 12.5 μmol/L) treatment for 24 h under FFA induction, the formation of intracellular lipid droplets was detected; Levels of triglyceride (TG) and total cholesterol (TC) were assessed using biochemical kits. DCFH-DA assay was utilized to measure intracellular reactive oxygen species (ROS) generation. Malondialdehyde (MDA), glutathione (GSH) levels and superoxide dismutase (SOD) activity as indicators of oxidative stress, were also determined. Western blotting was employed to detect the expressions of peroxisome proliferator-activated receptor α (PPARα), PPAR coactivator-1α (PGC-1α), and proteins involved in fatty acid oxidation, cholesterol metabolism, and nuclear factor erythrocyte 2-associated factor 2/heme oxygenase-1 (Nrf2/HO-1) pathway. Results Compared with control group, HepG2 steatosis cell model exhibited significantly increased intracellular TC, TG, ROS production and MDA levels (P < 0.001), alongside reduced SOD activity and GSH level (P < 0.01, 0.001). There was also a decrease in PPARα, PGC-1α, carnitine palmitoyl transferase 1A (CPT1A), acyl coenzyme A oxidase 1 (ACOX1), Nrf2 and HO-1 protein expressions (P < 0.05, 0.01, 0.001). Compared with model group, AE, alisol A and alisol A-23-acetate led to a significant reduction in intracellular lipid droplets, TG, and TC levels (P < 0.05, 0.01, 0.001); AE, alisol A and alisol A-24-acetate decreased ROS and MDA levels (P < 0.05, 0.001), increased SOD activity and GSH level (P < 0.05, 0.01). Furthermore, the expression levels of PPARα, PGC-1α, CPT1A, ACOX1, cytochrome P450 enzyme 7A1 (CYP7A1), Nrf2, and HO-1 proteins were significantly upregulated in each administration group (P < 0.05, 0.01). Conclusion AE, alisol A, alisol A-23-acetate and alisol A-24-acetate effectively mitigate lipid metabolism abnormalities and oxidative stress in FFA-induced HepG2 steatosis cell model. The mechanism maybe involve modulation of PPARα and Nrf2/HO-1 pathways, thus enhancing the fatty acid oxidation and cholesterol metabolism, and reducing oxidative stress.