祛湿化瘀方对游离脂肪酸诱导的HepG2细胞脂肪沉积和TNF-α分泌的抑制作用

Inhibitory Effects of Qushi Huayu Decoction on Fatty Deposition and Tumor Necrosis FactorαSecretion in HepG2 Cells Induced by Free Fatty Acid

目的研究祛湿化瘀方对体外肝脂毒性模型脂肪沉积和肿瘤坏死因子α(tumor necrosis factorα,TNF-α)分泌的影响,以进一步探究该方防治脂肪肝的作用机理方法复制游离脂肪酸(free fatty acid,FFA)诱导的HepG2细胞脂肪变性和TNF-α分泌脂毒性模型。设正常组、模型组和不同浓度药物血清组。分别观察上清中TNF-α含量,细胞内甘油三酯(triglyceride,TG)含量,细胞油红染色和电镜超微结构变化,细胞TNF-α蛋白表达及其基因表达。结果FFA刺激24 h后,模型组细胞内脂肪沉积明显,其TG含量显著高于正常组(P<0.01),同时上清中TNF-α含量、细胞内TNF-α的蛋白表达及其mRNA表达显著增强。而10%药物血清组细胞内TG含量及上清中TNF-α含量显著低于模型组(均P<0.01),其脂肪沉积、细胞内TNF-α的蛋白表达及其mRNA表达也明显轻于模型组。

Objective To study the effects of Qushi Huayu Decoction （QHD） contained serum on fatty deposition and tumor necrosis factor α （TNF-α） secretion of hepatic lipotoxicity model in vitro, for further investigating the mechanism of the decoction for preventing and treating fatty liver. Methods The steatosis with TNF-α secretion lipotoxic model of HepG2 induced by long-chain free fatty acid （FFA） was duplicated. Groups of normal, model cells and model cells treated with different concentrations of QHD contained serum were set up to test the content of TNF-α in culture supernate and triglyceride （TG） in cells, as well as to observe the ultrastructural change of cells by oil-red staining and the protein expression and gene expression of cellular TNF-α. Results After being stimulated with FFA for 24 h, marked deposition of lipid with high content of TG presented in the cells of model group. Compared with the normal group, not only TNF-α content of culture supernate but also the protein expression and mRNA expression of intracellular TNF-α increased significantly. Contrast to the model group, the contents of TG in cells and TNF-α in supernate as well as the protein and mRNA expression of TNF-α in the model cell group treated with 10 % QHD were lower significantly （all P 〈 0.01）. Conclusion QHD could significantly inhibit the fatty deposition and TNF-α secretion in HepG2 cells induced by free fatty acid.