枳椇4种成分对人肝细胞HL7702脂肪变性的抑制作用

Inhibition of Ethanol-induced Hepatic Steatosis in Human HL7702 Liver Cells by Four Constituents Isolated from Hovenia Acerba

为了筛检出枳椇中解酒保肝的有效成分,本文检讨了枳椇乙酸乙酯萃取物中的4种成分对肝细胞脂肪变性的影响。采用MTT法分析对照组、模型组和四种活性物高中低剂量组的细胞活性,并通过分析乙醇造模后油红O染色观察细胞内的脂滴情况,检测细胞TG以及肝酶ALT、AST的含量。通过MTT检测乙醇诱导细胞的最佳浓度发现,与对照组相比,0.9%乙醇造模48h后的细胞存活率下降,油红O观察乙醇造模后细胞内的脂滴情况更加明显,而且细胞肿胀变圆;四种活性物干预乙醇造模细胞后,与模型组相比,二氢杨梅素和槲皮素干预后,细胞的存活率略有提高;与对照组相比,模型组TG含量以及肝酶ALT、AST的泄漏量显著(p<0.05)升高。另外,与模型组相比,0.9%乙醇加二氢杨梅素或槲皮素共同作用正常人HL7702肝细胞48 h后,其TG以及肝酶ALT、AST的含量显著(p<0.05)下降。可见,初步确证枳椇乙酸乙酯层中具有解酒保肝作用的有效成分主要为二氢杨梅素和槲皮素,特别是前者。

To discover the effective substances in Hovenia acerba capable of dispeling the effects of alcohol and protecting the liver,this study examined the effects of four constituents in the ethyl acetate extract from Hovenia acerba on hepatic steatosis.The cells metabolic activity/viability of the control group,model group,or high-/medium-/low-dose group of the four bioactives was measured by the MTT assay. The lipid droplets in the cells were observed by Oil red O staining after ethanol modeling.The levels of cell TG and liver enzymes,ALT and AST,were determined.The optimal concentration of ethanol-induced cells was measured by the MTT assay.It was found that compared with the control group,the cell viability decreased after of0.9%ethanol modeling for48h,along with the observations that Oil red O-stained lipid droplets became more obvious in the ceils and the cells became swollen and round,after the exposure of ethanol-treated model cells to each of the four bioactives,a slight increase in the survival rate of the cells was induced by dihydromyricetin and quercetin as compared with the model group;compared with the control group,the TG content and the leakage of ALT and AST of the model group significantly increased(p<0.05). Further,compared with the model group,the application of0.9%ethanol combined with dihydromyricetin or quercetin on the normal human HL7702hepatocytes for48h caused a significant(p<0.05)decrease in the TG and ALT,AST contents in the liver.Accordingly,the active ingredients in the ethyl acetate phase obtained from Hovenia acerba that were capable of dispeling the effects of alcohol and protecting the liver were mainly dihydromyricetin and quercetin,especially the former.