雌激素受体介导Genistein对内皮型一氧化氮合酶表达的诱导作用

The Action of Estrogen Receptor in Genistein Upregulating the Expression of Endothelial Nitric Oxide Synthase

目的研究雌激素受体介导的基因组效应在Genistein促进内皮型一氧化氮合酶表达过程中的作用。方法体外培养人脐静脉内皮细胞,在氧化型低密度脂蛋白(100mg/L)干预内皮细胞的基础上,经雌激素受体拮抗剂ICI182780(1μmol/L)或经基因转录阻断剂放线菌素D(5mg/L)作用30min后,再加入Genistein(100nmol/L)作用24h,实时定量PCR检测内皮型一氧化氮合酶mRNA表达,WesternBlotting检测内皮型一氧化氮合酶蛋白的表达。结果与空白对照组相比,氧化型低密度脂蛋白(100mg/L)明显下调内皮型一氧化氮合酶mRNA和蛋白表达(P<0.05);Genistein明显上调内皮型一氧化氮合酶mRNA和蛋白表达(P<0.05),而且Genistein对内皮型一氧化氮合酶的诱导作用被雌激素受体拮抗剂ICI182780和基因转录阻断剂放线菌素D明显抑制(P<0.05)。结论 Genistein促进内皮型一氧化氮合酶的表达与雌激素受体介导的基因组效应密切相关。

Aim To investigate the action of estrogen receptor in Genistein upregulating the expression of endothelial nitric oxide synthase（eNOS）in endothelial cells.Methods Human umbilical vein endothelial cells were exposed to medium or oxide low density lipoprotein（ox-LDL）（100 mg/L）in the presense or absence of estrogen antagonists ICI182780（1 μmol/L）or/and actinomycin D（5 mg/L）for 0.5 hour,then were treated with Genistein（100 nmol/L）for 24 hours.The mRNA expression of eNOS was detected by real-time PCR,the protein expression of eNOS was determined by Western Blotting.Results ox-LDL downregulated the expression of eNOS mRNA and protein（P〈0.05 vs that of basal level）;Genistein upregulated the expression of eNOS mRNA and protein（P〈0.05 vs that of ox-LDL-treated group）,furthermore,the increase can be inverted by estrogen antagonists ICI182780 or actinomycin D（P〈0.05）.Conclusion Genistein could upregulate the expression of eNOS,which is associated with estrogen receptor regulating genes transcription.