过氧化物酶体增殖物激活受体δ下调载脂蛋白F在肝细胞中的表达

Peroxisome Proliferator Activated Receptor Delta Agonist Downregulates the Expression of Apolipoprotein F in Liver Cells

目的探讨过氧化物酶体增殖物激活受体δ(PPARδ)对载脂蛋白F(ApoF)基因转录的调控作用及机制。方法使用不同浓度PPARδ激动剂GW0742干预HepG2细胞株,Realtime PCR检测ApoF的mRNA水平,Western-blot检测ApoF的蛋白水平;构建ApoF启动子双荧光素酶报告基因重组载体,与PPAR激动剂共转染肝细胞;双荧光素酶报告基因系统检测PPARδ激动剂对启动子活性的影响。结果经不同浓度PPARδ激动剂GW0742处理后,10μmol/L以上浓度的ApoF mRNA及蛋白水平显著低于未处理组;PPARδ激动剂可以显著降低ApoF的启动子活性。结论PPARδ可以通过调控ApoF启动子的活性下调其mRNA及蛋白的表达水平。

Objective Apolipoprotein F regulates the activity of cholesterol ester transporters.Previous studies have found that its upstream promoter region is regulated by ETS and CEP transcription,and further studies on the role and possible mechanism of PPAR in apolipoprotein F are proposed.Methods Using different concentrations of PPARδagonists(GW0742)to intervene the cell lines to detect apolipoprotein F mRNA levels by Realtime-PCR,in addition,the upstream promoter of double fluorescent reporter enzyme apolipoprotein was constructed by gene cloning and co-transfected into hepatocytes with PPAR agonists.The effect of PPARδagonists on promoter activity was detected by double fluorescent reporter enzyme gene.Results The ApoF mRNA level of PPARδgroup with concentration above 10μmol was significantly lower than that of untreated group after treated with different concentrations of agonists.PPARδagonists can significantly reduce the activity of ApoF promoter.Conclusion PPARδcan down-regulated the expression level of Apo-F protein and mRNA by regulating the activity of ApoF promoter.