血管平滑肌细胞亲环素A受体克隆鉴定及功能研究

Inflammation and oxidative stress are pathogenic mediators of many cardiovascular diseases, but therapeutic approaches targeting these mediators have not been effective. Recently, it was reported that cyclophilin A (CypA) as a secreted oxidative stress induced factor (SOXF) was released from vascular smooth muscle cells (VSMC) in response to reactive oxygen species (ROS), and mediated extracellular signal-regulated kinase1/2 (ERK1/2) activation and VSMC growth. However the unequivocal identity of the CypA receptor(eCypAR)remains unknown. .Several molecules have been proposed as the eCypAR including CD147, syndecan-1 and very late activation antigen-5. However, our preliminary data suggest that these molecules are unlikely to be the eCypAR in VSMC and EC.Therefore, in disagreement with the current literature, we hypothesize that a high affinity receptor exists for eCypA in VSMC and in this study propose to identify and characterize it by using ligand-receptor binding, isolation, purification and MS analysis as well as overexpression, knockdown and functional rescue of candidate receptor. Once eCypAR is identified, it will provide clues toward probing currently unknown functional mechanisms of eCypA and signal transduction in the above diseases and may lead to identification of novel therapeutic targets.

炎症和氧化应激是许多心血管疾病的主要致病机制，但目前仍缺乏针对性的有效治疗手段。我们前期研究发现胞外亲环素A(eCypA)作为一种新型的分泌型氧化应激诱导因子介导了心血管疾病中炎症和氧化应激的病理过程，但其受体并非已报导的几种分子如CD147、多配体（蛋白）聚糖1和晚期激活抗原5等。本课题旨在：通过配基-受体结合、分离、纯化、质谱分析的前向遗传克隆策略筛选eCypA受体（eCypAR），并通过候选受体过表达、表达抑制、功能缺失-获得技术进一步确定eCypAR。鉴定eCypAR有助于阐明eCypA及信号转导通路在心血管病变中的作用，为筛选新的治疗靶点提供理论依据。

炎症和氧化应激是许多心血管疾病的主要致病机制，但目前仍缺乏针对性的有效治疗手段。研究发现胞外亲环素A(eCypA)作为一种新型的分泌型氧化应激诱导因子介导了心血管疾病中炎症和氧化应激的病理过程，但其受体并非已报导的几种分子如CD147、多配体（蛋白）聚糖1和晚期激活抗原5等。我们构建并表达纯化了CypA蛋白，并通过配基-受体结合、分离、纯化、质谱分析的前向遗传克隆策略筛选了eCypA受体（eCypAR），将通过候选受体过表达、表达抑制、功能缺失-获得技术进一步确定eCypAR。鉴定eCypAR有助于阐明eCypA及信号转导通路在心血管病变中的作用，为筛选新的治疗靶点提供理论依据。同时，启动了与CypA相关的其他课题。