【摘要】 目的探讨胞内S-腺苷同型半胱氨酸(SAH)升高抑制血管内皮细胞增殖和雌激素受体-α(ER-α)表达变化的关系。方法以永生化的人脐静脉内皮细胞(HUVEC)为研究对象,经不同浓度的S-腺苷同型半胱氨酸水解酶(SAHH)强效抑制剂3-deazaadenosine(DZA)处理24,48,72 h。利用细胞计数法、流式细胞仪和脱氧核糖核酸转移酶缺口末端标记法(TUNEL)检测细胞的增殖凋亡能力,蛋白印迹法检测ER-α蛋白的表达,荧光定量-PCR(qRT-PCR)检测其mRNA表达,甲基化特异PCR法(MSP)检测ER-α基因启动子甲基化状态。结果经DZA处理后,HU-VEC增殖能力降低(主要受阻于G1/G0期),细胞凋亡率增加(P<0.05),ER-αmRNA和蛋白的相对表达量降低(P<0.05),但其启动子甲基化不发生改变。结论胞内SAH升高抑制HUVEC的增殖能力与ER-α的表达变化有关,但这种作用不是通过改变ER-α基因启动子的甲基化而实现。更多还原

【Abstract】 Objective To explore the relationship of inhibitory proliferative ability induced by intracellular S-adenosylhomocysteine(SAH) accumulation and the expression of estrogen receptor-α(ER-α) in human umbilical vein endothelial cells.Methods Immortalized HUVEC were treated without(normal) or with different concentration 3-deazaadenosine(DZA),a potent S-adenosylhomocysteine hydrolase(SAHH) inhibitor,for 24,48 and 72h.The proliferative ability of HUVEC was measured with flow cytometry and TdT mediated dUTP nick ending labelling(TUNEL) methods.The ER-α protein and mRNA expression was analyzed with western blot and quantitative reverse transcription-polymerase chain raction(qRT-PCR) respectively.The promoter methylation of ER-α gene was detected with methylation special PCR(MSP).Results The proliferative ability was decreased(mainly arrested at G1/G0 phase),and the apoptotic rate was increased after HUVEC treatment with DZA(P<0.05).The relative expression of ER-α mRNA and protein in HUVEC treated with DZA was lower than that of normal(P<0.05),but there was no changes of ER-α gene promoter methylation.Conclusion The inhibitory proliferative ability induced by intracellular SAH accumulation is correlative with ER-α expression,which not through altering promoter methylation in HUVEC.更多还原