内皮源性endophilinA2整合雌激素基因和非基因途径的机制及其抗动脉粥样硬化的作用研究

Estrogen has been proved to inverse vascular endothelial dysfunction, which is elicited by estrogen’s genomic and nongenomic pathways. There is integration between these two pathways, while the integrative mechanisms remain obscure. Early results had demonstrated that Endophilin A2 (Endo II) plays critical role in the regulation of vascular function. Our preliminary results showed that 17β-estradiol (E2) enhanced EndoII mRNA and protein expression and the function of E2 to induce rapid vasodilation effects disappeared after silencing Endo II.on the basis of these evidences, we proposed the following hypothesis that EndoII mediate E2’s against atherogenesis effect by serving as the critical molecule to integrate estrogen’s genomic and nongenomic pathways. Therefore, the present study aims to investigate the genomic mechanism of E2 to increase EndoII protein expression; the key role of EndoII as a node molecule in maintaining the homeostasis of nongenomic pathways will be clarified; furthermore, we will explore the intergrating effects of Endo II in improving endothelial function; finally, we will use the double knockout mice (ApoE-/-EndoII-/-) model to observe the role of EndoII in estrogen’s anti-atherogenesis effect. The present project aims to explore the molecular mechanism of estrogen’s anti-atherogenesis effect from the new perspective of integrating node molecules, clarify the role of EndoII in integrating the E2’s genomic and nongenomic effects,which will shed light on the discovery of new target for the administration of estrogen replacement therapy on the prevention of cardiovascular diseases.

雌激素逆转绝经后血管内皮功能失调，其机制涉及基因和非基因途径，两者存在整合效应，但整合机制不明。前期结果显示，EndophilinA2（EndoII）在调控血管功能中发挥重要作用。预实验结果表明，17β-雌二醇（E2）上调内皮EndoII表达；沉默EndoII后E2快速舒张血管作用消失。由此提出科学问题：EndoII是否作为节点分子，整合基因和非基因途径，介导E2抗动脉粥样硬化（AS）效应？为此，本项目拟探索E2上调EndoII表达的基因机制；阐明EndoII作为节点分子维持非基因途径稳态的关键作用；进一步探索其整合效应在改善内皮功能中的作用；最终在EndoII联合ApoE基因敲除小鼠AS模型中，揭示此整合机制在延缓AS中的作用。本研究从整合节点分子这一新视角揭示雌激素抗AS的分子机制，阐明EndoII对基因和非基因途径的整合作用，为雌激素替代疗法应用于绝经后心血管疾病防治提供新干预靶点。

雌激素逆转绝经后血管内皮功能失调，其机制涉及基因和非基因途径，两者存在整合效应，但整合机制不明。前期结果显示，EndophilinA2（EndoII）在调控血管功能中发挥重要作用。本项目主要（1）观察雌激素对内皮EndoII的调控效应并探讨其调控EndoII的分子机制。（2）探讨EndoII作为整合节点分子稳定非基因信号分子ERα的机制。（3）研究EndoII在雌激素改善内皮功能的关键作用。研究结果表明，雌激素通过基因效应促进EndoII的mRNA和蛋白质表达，其机制为ERα与EndoII的ERE反应元件结合，促进其转录。此外，短时间雌激素处理脐静脉内皮细胞，增加pTyr-EndoII水平，减弱EndoII对ERα的内吞，使细胞膜上ERα增加，促进mERα介导的血管舒张。当沉默EndoII后，E2诱导的上述效应进一步增加。本项目首次揭示EndoII作为整合节点分子，在整合雌激素基因和非基因途径的关键作用。提示EndoII对雌激素的基因效应和非基因效应有协作作用，通过EndoII对对雌激素非基因效应进行选择性干预，可为开发新型选择性雌激素受体调节剂以防治AS提供新的策略。