脱氢表雄酮通过抑制高脂诱导的巨噬细胞炎性蛋白-1α抗动脉粥样硬化的机制

Mechanism of dehydroepiandrosterone resisting atherosclerosis by inhibiting the expression of MIP-1α induced by high lipid

目的:探讨脱氢表雄酮(DHEA)是否通过抑制高脂诱导的巨噬细胞炎性蛋白-1α(MIP-1α)的表达发挥抗动脉粥样硬化(AS)作用,以及细胞色素P450芳香酶基因(CYP19)对DHEA的作用。方法:氧化低密度脂蛋白(ox-LDL)诱导人脐静脉内皮细胞(HUVEC),给予DHEA、全反式维甲酸(ATRA)干预;pcDNA3.1-CYP19-GFP真核表达质粒及pcDNA3.1-GFP空质粒分别转染HUVEC,给予ox-LDL诱导及DHEA干预。实时荧光定量PCR和细胞酶联免疫吸附法检测各组HUVEC及各组转染细胞MIP-1αmRNA及蛋白的表达。高脂饮食建立AS兔模型,给予DHEA、ATRA干预,实时荧光定量PCR和免疫组织化学法检测兔主动脉MIP-1αmRNA及蛋白的表达。结果:ox-LDL诱导的HUVEC的MIP-1α表达显著升高,给予DHEA干预后,MIP-1α的表达明显回降(P均<0.05),同时给予ATRA干预对DHEA抑制MIP-1α的表达无明显影响。喂饲高脂饲料的大耳白兔,主动脉MIP-1α表达显著升高,而在高脂饲料中添加DHEA后,MIP-1α的表达明显回降(P均<0.05),同时添加ATRA对DHEA抑制MIP-1α的表达无明显影响。经ox-LDL诱导及DHEA干预后,转染CYP19质粒的HUVEC MIP-1α的表达较转染空质粒的HUVEC显著降低(P<0.05)。结论:DHEA能够抑制高脂诱导的HUVEC和兔主动脉MIP-1α的表达,发挥抗AS作用;CYP19能够增强DHEA的作用。

Objective：To investigate whether dehydroepiandrosterone（DHEA）can resist atherosclerosis by inhibiting the expression of macrophage inflammatory protein-1α（MIP-1α）induced by high lipid,and the effect of cytochrome P450 aromatase gene（CYP19）on DHEA. Methods：Human umbilical venous endothelial cells（HUVEC）induced by ox-LDL were intervened by DHEA and ATRA.The eukaryotic expression plasmid pcDNA3.1-CYP19-GFP and pcDNA3.1-GFP were transfected into HUVEC respectively.The transfected HUVEC were intervened by ox-LDL and DHEA.The expression of MIP-1αmRNA and protein in all groups of HUVEC and transfected HUVEC were determined by RT-qPCR and cell ELISA.Rabbit atherosclerosis model caused by high lipid diets were intervened by DHEA and ATRA.The expression of aortic MIP-1αmRNA and protein were determined by RT-qPCR and immunohistochemistry. Results：Ox-LDL stimulation significantly induced MIP-1αexpression in HUVEC（P〈0.05）.After intervened by DHEA,the MIP-1αexpression was obviously decreased（P〈0.05）.ATRA had no remarkable effect on MIP-1αexpression inhibited by DHEA in HUVEC（P〉0.05）.High lipid diets significantly induced MIP-1αexpression in rabbit aorta（P〈0.05）.After DHEA was added,the MIP-1αexpression was obviously decreased（P〈0.05）.ATRA also had no remarkable effect on MIP-1αexpression inhibited by DHEA in rabbit aorta（P〉0.05）.After intervened by ox-LDL and DHEA,the MIP-1αexpression in CYP19 plasmid transfected HUVEC was significantly decreased compared with empty plasmid transfected HUVEC. Conclusions：DHEA resist atherosclerosis by inhibiting the expression of MIP-1αinduced by high lipid in HUVEC and rabbit aorta.CYP19 can enhance the effect of DHEA.