建立非酒精性脂肪肝细胞模型,研究茯茶中“金花”孢子粉提取物对非酒精性脂肪肝细胞甘油三酯代谢的影响,筛选出具有降脂功效的活性组分,为茯茶的深度开发及“金花”孢子粉降脂成分的分离纯化提供一定的指导。以正常人肝细胞LO2为实验对象,采用脂肪乳剂加入细胞培养液培养肝细胞,建立非酒精性脂肪肝细胞模型;以MTT法筛选出提取物最佳作用浓度;通过检测细胞内甘油三酯来评价药物的降脂效果。结果表明,用含1.0mL/L 20%脂肪乳剂的培养液培养LO2肝细胞48h,肝细胞内甘油三酯与对照组比较呈现极显著升高（P<0.01）且肝细胞ALT、AST泄漏量比对照组极显著加大（P<0.01）,因此本方法能建立非酒精性脂肪肝细胞模型;“金花”孢子粉提取物作用脂肪肝细胞最佳质量浓度分别为150μg/mL和5μg/mL,当醇提物和萃取的水层部分分别作用脂肪肝细胞24、48h时都能使细胞内甘油三酯含量降低且与阳性药物辛伐他汀和血脂康效果相当,因此本实验筛选出“金花”孢子粉醇提物和萃取的水层具有较好的降脂效果。
To establish cellular models of non-alcoholic fatty liver(NAFL), study the effects of jinhua spores extracts on metabolizability of intracellular triglyceride and screen out the effective lipid-lowering components, for providing the guidance in investigating the comprehensive development of Fu tea, separation and purification of lipid-lowering components of jinhua spores. Liver cells LO2 were cultured with medium containing fat emulsion to establish cellular models of NAFL. MTT assay was used to screen out the optimal concentration of extracts. Lipid-lowering effects of extracts were evaluated by intracellular triglyceride level. Results showed that cellular models of NAFL were cultured in 48 hours by mixing medium, which contains 1.0mL/L 20% fat emulsion. Compared with controls, triglyceride in liver cells, leakage of liver cells ALT and AST showed extremely significant rise(P<0.01). So this method can be used to establish cellular models of NAFL. In addition, Jihua spores extrcacts had some inhibitory effects on NAFL cells and the optimal concentration respectively was 150μg/mL and 5μg/mL. When fatty liver cells were stimulated 24h, 48h by ethanol extracts and extracts of water layer, the intracellular triglyceride levels can be reduced, and the positive effects smiliar to Simvastatin and Xuezhikang. Obviously, the ethanol extractions and extractions of warter layer possessed good lipid-lowering effects.
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