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.
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)
