毛菊苣有效部位降脂作用及初步机制探讨

摘要

Objective To explore lipid-lowering effect of the effective parts of Cichorium glundulosum and its preliminary mechanism. Methods MTT method was used to determine the amount of serum,as well as Nile red staining to establish hyperlipidemia model of HepG2 cells.On this basis,the content of key index TG and the lipid droplets were determined.Finally,ELISA enzyme-linked immu-nosorbent assay was used to make a preliminary study of the lipid-lowering mechanism of Cichorium glundulosum effective parts from the aspects of the absorption,oxidation and TG synthesis of free fatty acid.Results It was showed that,through MTT experiment and Nile red staining and the flow type quantitative analysis,fat drops content of module cell was obviously increased more than that of con-trol group and had significant difference (P <0.01 and 0.001)when fat breast concentration was 6% of the total volume;content of TG (P <0.05)and fat drops (P <0.01 and 0.001)of HepG2 cells were significantly reduced in different models of rat,and medicated serum in normal state had more significant lipid-lowering effect than that in pathological state.And tests of the three indexes GPAT1 -2 (P <0.05),PPARγ(P <0.05 and 0.01),FATP5(P <0.01)showed that there was significant difference between medicated serum in normal state group and that in high lipid model group.Conclusions Medicated serum with the effective parts of Cichorium glundulo-sum in pathological state (high lipid model)or in normal state can both significantly lower hyperlipidemia model of human hepatoma cell line HepG2 cells and the latter had more significant lipid-lowering effect,the mechanism of which may be related to reducing the content of GPAT1 -2 ,promoting the expression of PPARγand inhibiting the FATP5 generation.%目的：在整体水平上，研究毛菊苣有效部位含药血清降脂作用及初步机制。方法利用 MTT 法确定血清的加入量以及尼罗红染色建立 HepG2细胞高脂模型，在此基础上，检测细胞内的关键性指标甘油三酯（TG）和脂滴的含量，最后通过ELISA 酶联免疫吸附法分别从游离脂肪酸的吸收、氧化、TG 的合成方面对毛菊苣降脂有效部位含药血清的降脂机制进行初步的探讨。结果通过 MTT 实验和尼罗红染色及流式定量分析表明，脂肪乳浓度为总体积的6％时，造模组细胞内的脂滴含量较正常对照组明显增加，具有显著性差异（P ＜0．01、0．001）；不同模型的大鼠含药血清均可显著性地降低 HepG2细胞内 TG（P＜0．05）和脂滴（P ＜0．01、0．001）的含量，并且正常状态下的含药血清比病理状态下的含药血清降脂效果显著；同时对 TG 合成、氧化、吸收通路中的关键酶 GPAT1－2（P ＜0．05）、PPARγ（P ＜0．05、0．01）、FATP5（P ＜0．01）三个指标上的检测表明正常状态下的含药血清与高脂模型组相比具有显著性差异。结论大鼠病理状态下（高脂模型）与正常生理状态下的毛菊苣降脂有效部位含药血清具有显著性地降低人肝癌 HepG2细胞高脂模型的作用，并且后者较前者降脂效果明显，其机制可能与减少GPAT1－2的含量、促进 PPARγ的表达和抑制 FATP5的生成有关。