Effect Of Oleoylethanolamide On Fatty Liver Cell Model And Its Mechanis

Objective: on the basis of established fatty liver model in vitro to explore the mechanism and function of OEA treatment in fatty liver cell model in vitro.Methods: Experimental study based on HepG2 cell that were incubated with different concentrations of medical fat emulsion(MFE) and 10%fetal bovine serum for 24 hours to establish the fatty liver model in vitrol. To determine the best concentration for the model , MTT(thiazolyl blue) method was used the to measuring the activeity of cell, and red Oil O staining to observing the lipid drops in cells, and commercial clinical diagnosis of kit to detecting triglyceride content of cell lysis supernatant. Established fatty liver model treated with OEA and Fenofibrate as positive control for 6,12,24hours.After OEA administered, mRNA was extracted by Trizol Reagent and cDNA was synthesized by Superscriptase II. The levels of gene expression were measured by real time PCR. The change of lipid drop and triglyceride content in HepG2 cells were observed by red Oil O staining and commercial clinical diagnosis of kit, respectively.Results: Steatosis of the HepG2 cell was improved by OEA or fenofibrate treatment and intracellular triglyceride levels were significantly decreased by OEA with 10μM and 50μM (5.5 and 6.5-fold, p<0.001, respectively), which showed by Oil red O staining and the measurement of triglyceride. Furthermore, expresstion of the gene controlling fatty acid uptake, transport, oxidation and post-inflammation was changed. the mRNA expression of PPAR-α, Sirt-1 and carnitine palmitoyltransferase-1 (CPT-1) were increased in OEA treatment(P<0.05). In contrast, SREBP-1c, the gene preferentially regulate fatty acid synthesis, were markedly suppressed in OEA treatment(P <0.001) , and the mRNA expression of TNF-αwas also dramatically decreased in OEA treatment(P <0.001).Conclusions: HepG2 cell were induced steatosis by cultured with 1%MFE 24h, which can be the fatty liver cell model in vitro. OEA contributes to lower triglycerides and lipid accumulation in fatty liver cell model by inducing the expression of fatty-acid metabolic genes, such as PPAR-α, Sirt-1, and CPT-1. In addition, the effect of OEA on reduction of liver steatosis in vitro may be related to the decrease of SREBP-1c and TNF-αexpression implying the property of OEA to alleviate the fatty liver in cell model.