中文摘要：

为研究AP-1和STAT5协同调节血管紧张素原基因表达的作用机制,用凝胶电泳迁移率改变分析（EMSA）和染色质免疫沉淀分析（ChIP assay）检测血管紧张素Ⅱ（AngⅡ）诱导的AP-1和STAT5的DNA结合活性;用凝胶超迁移分析和免疫共沉淀方法观察AP-1和STAT5的相互作用.结果显示,AngⅡ可分别促进AP-1和STAT5与血管紧张素原基因调控区顺式元件的结合以及二者之间的相互作用;核蛋白与含AP-1结合位点的寡核苷酸探针结合形成的复合物可与抗c-Jun抗体和抗STAT5抗体形成超迁移电泳区带;而用抗c-Jun抗体也可从STAT5-染色质免疫沉淀复合物洗脱液中检测到AP-1的存在.而且,AP-1和STAT5的相互作用程度及其二者与DNA的结合活性与血管紧张素原表达活性具有一致关系,该效应可被JAK2特异抑制剂AG490所抑制.上述结果提示,与顺式元件结合的AP-1和STAT5通过相互作用而形成四元复合物协同调节血管紧张素原基因的表达,JAK2对该过程具有诱导活化作用.

英文摘要：

AP-1 and STATS have been shown to play a critical role in transcription of angiotensinogen gene in VSMC. To determine the mechanism of AP-1 and STATS cooperative regulation, the binding activity of AP-1 and STATS to cis-acting elements located in angiotensinogen gene was detected by EMSA and CHIP, respectively, following Ang Ⅱ induction. The interaction of AP-1 with STATS was demonstrated using supershifted assay and immunoprecipitation. The results showed that Ang Ⅱ stimulated AP-1 or STATS binding to DNA element and the interaction between two factors. The supershifted bands could be formed in EMSA using AP-1 binding site as probe after adding antibody AP-1 or anti- STATS. It was found that AP-1 existed in the immunoprecipited complex of STATS-chromatin by Western blotting. The interaction of AP-1 with STATS was associated with them binding to cis-acting elements and expression of angiotensinogen gene. These findings suggest that AP-1 and STATS bound to cis-elements regulate cooperatively angiotensinogen gene transcription through interaction between them.