| In the experiment, human hepatocellular carcinoma HepG2 and human hepatocyte L02 was selected as experimental cell and the model of steatotic cells in vitro was constructed by oleic acid (OA). The SPO PUFAs and ALA which is the main ingredient of SPO PUFAs were chosen as the tested substances to study the effects of SPO PUFAs and ALA on lipid accumulation, cholesterol metabolism and transformation for HepG2 and L02 cells under fatty change. To explore the effects of SPO PUFAs and ALA on cholesterol efflux in HepG2 and L02 cells via LXRa/PPARy-ABCA1/ABCG1-CYP7A1 pathway, real-time PCR was used to analyse the expression levels of LXR?, PPARy, ABCA1, ABCG1 and CYP7A1 mRNA, and western-blot was used to measure the protein expression of them. The results we had found are as follows:(1)SPO PUFAs and ALA could suppress lipid accumulation of HepG2 and L02 cells induced by OA. Intracellular TC was markedly decreased and TBA contents in the medium were dose-dependently increased with tested substance. This showed that SPO PUFAs and ALA could increase cholesterol removal and then transform bile acid.(2)The result of real-time PCR and western-blot showed that the relative mRNA and protein expression of LXR?, PPARy and their target genes ABCA1. ABCG1 and CYP7A1 could be up-regulated by SPO PUFAs and ALA as well as exhibited a dose-dependent manner.This showed the molecular mechanism of SPO PUFAs and ALA promoting cholesterol metabolism and transformation, inhibiting hepatocellular lipid change in liver cells may be LXRa/PPARy-ABCA1/ABCG1-CYP7A1 cell signaling pathways.(3)Taken all the results together, we found the effect of SPO PUFAs were better than ALA on cholesterol metabolism and transformation x.(4)The results of HepG2 were similar to L02 cells in the experiment, which showed human hepatocellular carcinoma HepG2 can also correctly reflect the cholesterol metabolism affected by food functional components of liver cells. |
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