长链脂肪酸对脂周素2表达的影响

The effect of long-chain fatty acids on the expression of perilipin 2 in NMuLi cells

目的研究长链脂肪酸对肝细胞脂周素2表达的影响,并揭示其调控的相关分子机制。方法应用实时定量PCR测定不同长度脂肪酸对脂周素2mRNA表达的影响。应用实时定量PCR和West-ernblot分别测定长链脂肪酸油酸对脂周素2mRNA和蛋白表达的影响。应用荧光素酶活性分析方法检测油酸对脂周素2启动子活性的影响。结果长链脂肪酸油酸、软脂酸、亚油酸、亚麻酸和花生四烯酸明显增加了脂周素2的表达,而短链脂肪酸己酸没有作用。油酸以剂量和浓度依赖方式上调脂周素2mRNA和蛋白的表达,乙酰辅酶A合成酶抑制剂triacsinC没有抑制油酸诱导的脂周素2表达。小鼠脂周素2启动子含有Ets/AP-1结合位点和PPARs反应元件(PPRE)。油酸以剂量依赖的方式显著增强了脂周素2的-2090bp启动子活性。Ets位点突变没有影响脂周素2的启动子活性,但AP-1位点突变,及PPRE突变显著抑制了油酸诱导的启动子活性。结论在肝细胞中,长链脂肪酸油酸诱导脂周素2的表达,需要启动子区域的AP-1和PPARs反应元件,为防治脂肪肝新药研发提供了新的靶点。

Objective To investigate the effect of long-chain fatty acids on perilipin 2 expression in NMuLi cells. Methods Real-time PCR and Western blot were performed to detect perilipin 2 expression. Transient transfection and luciferase assay were employed to measure the perilipin 2 promoter activity. Results Long-chain fatty acids enhanced perilipin 2 expression, but not short-chain fatty acid. Oleic acid significantly induced perilipin 2 expression in a dose and time dependent manner in NMuLi cells. Oleic acid markedly enhanced perilipin 2 promoter activity, and this process required AP-1 and PPARs activity. Conclusions These studies suggest new information on the regulation of perilipin 2 in NMuLi cells,which provide a new target for prevention or therapy of fatty liver.