植物雌激素α-ZAL抑制同型半胱氨酸诱导HUVECs的ET-1基因表达及抗氧化机制  

Phytoestrogen α-Zearalanol Inhibits HCY-Induced Endothelin-1 Gene Expression and Mechanism of HUVECs

作　　者：段金虹[1] 徐海珊[1] 戴顺龄[1] 王小明[1] 吴云清[1] 张彦东[1] 孙仁宇[1] 

机构地区：[1]中国医学科学院基础医学研究所中国协和医科大学基础医学院病理生理学系,邮政编码北京100005

出　　处：《微循环学杂志》2006年第3期11-15,共5页Chinese Journal of Microcirculation

基　　金：国家自然科学基金项目(30170376)

摘　　要：目的:通过氧应激(ROS)信号转导通路探讨植物雌激素α-ZAL对同型半胱氨酸(HCY)诱导人脐静脉内皮细胞(HUVECs)中ET-1基因表达的抑制作用及抗氧化机制。方法:用荧光探针DCF检测HUVECs内ROS的含量;采用RT-PCR方法对ET-1mRNA的表达进行分析,运用WesternBlot法测定细胞外信号调节激酶(ERK)、磷酸化细胞外信号调节激酶(pERK)和c-Jun氨基末端激酶(c-jun/AP-1)的表达;同时测定HUVECs上清液中ET-1的浓度,利用瞬时转染技术观察HUVECs的AP-1报告基因的活性。结果:α-ZAL能降低AP-1的活性,抑制由HCY诱导ET-1的分泌和ET-1mRNA的表达,并可能通过抗氧化作用阻断由HCY诱导的HUVECsROS的产生。结论:α-ZAL能抑制HCY诱导HU-VECs的ET-1基因表达,推测可能是α-ZAL减弱了氧应激(ROS)、抑制了蛋白激酶ERK的激活和调节ET-1基因上核转录因子AP-1的表达。Objective: To examine the inhibitory effect and antioxidative mechanism of phytoestrogen α-Zearalanol(α-ZAL)on Homocysteine(HCY)-induced ET-1gene expression through ROS mediated signal transduction pathways in HUVECs.Method: Using the fluorescent probe DCF,the effects of α-ZAL on HCY -induced redox level in HUVECs were determined. The influences of α-ZAL on expression of mRNA of ET-1 and expressions of protein of ERK, pERK and c-jun/AP-1 induced by HCY in HUVECs were measured using RT-PCR and Western Blotting respectivety. The effect of α-ZAL on ET-1 secretion level induced by HCY were determined in HUVECs using enzymatic immunoassay. We performed transient transfection assay and the effects of α-ZAL on transcriptional factor AP-1induced HCY were detected.Results: The α-ZAL could inhibit the secretion of ET-1, expression of ET-1mRNA and oxidative stress induced by HCY, and downregulate enhanced of AP-1 activity and expression induced by HCY.Conclusion: The results suggest that α-ZAL inhibited the effects of HCY on promoting the expression of ET-1 gene through attenuating ROS formation and inhibiting the activation of ERK and expression of nuclear factor.

关 键 词：同型半胱氨酸 HUVECS 抗氧化机制 基因表达 ET-1 雌激素Α 动脉粥样硬化(AS) 内皮功能紊乱 血管平滑肌细胞 植物 

分 类 号：R363[医药卫生—病理学]