脂肪酸结合蛋白4 DNA甲基化在L-NAME干预胎盘滋养细胞脂代谢异常中的作用

Effect of FABP4 DNA methylation on abnormal lipid metabolism in trophocyte treated with L-NAME

目的观察并分析一氧化氮合酶抑制剂L-NAME对滋养细胞脂肪酸结合蛋白4(FABP4)基因启动子区DNA甲基化及脂质代谢异常的影响。方法体外培养人胎盘滋养细胞(HTR-8)并用100μmol/L-NAME干预48 h后检测胎盘滋养细胞脂质含量;qRT-PCR和Western blot检测胎盘滋养细胞中DNA甲基转移酶1(DNMT1)、FABP4 m RNA和蛋白表达改变;巢式甲基化特异性PCR(NT-MSP)检测胎盘滋养细胞中FABP4启动子区DNA甲基化的程度改变。结果 L-NAME干预滋养细胞后脂质含量明显增多,且细胞内FABP4 mRNA及蛋白表达显著增加(P<0.05),而FABP4基因启动子区甲基化程度以及DNMT1的表达水平显著减低。结论 FABP4启动子区低DNA甲基化可能参与了L-NAME诱导的胎盘滋养细胞脂代谢异常的调控过程。

Objective To investigate the effect of fatty acid binding protein4(FABP4)DNA methylation on abnormal lipid metabolism in placental trophoblastic dyslipidemia.Methods Human placental trophoblast cell line(HTR?8)was treated with L-NAME of100μmol/L for48h.The lipid content in placental trophoblasts was detected by chemical enzyme-colorimetry.The FABP4DNA methylation level in placenta trophoblasts was detected by nested?touch down methylation specific PCR(NT-MSP).the mRNA and protein expression of DNMT1and FABP4were detected by qRT-PCR and Western Blot,respectively,in trophoblast cells.Results The lipid contentin trophoblasts significantly increased as compared with the control(P<0.05).Expression of FABP4mRNA and protein increased(P<0.05),while FABP4methylation level and expression of DNMT1significantly decreased(P<0.05)after treatment with L-NAME.Conclusions FABP4DNA methylation is involved in the regulation of lipid metabolism in placental trophoblastic cells of hypertensive disorder complicating pregnancy.