肽基脯氨酰异构酶1在2型糖尿病下肢动脉再狭窄中的作用及机理研究

项目名称： 肽基脯氨酰异构酶1在2型糖尿病下肢动脉再狭窄中的作用及机理研究

项目编号： No.81200227

项目类型： 青年科学基金项目

立项/批准年度： 2013

项目学科： 医学一处

项目作者： 吕磊

作者单位： 上海交通大学

项目金额： 23万元

中文摘要： 血管腔内治疗术是目前临床上治疗下肢动脉硬化闭塞症的重要方法。如何降低术后再狭窄率是当今医学研究的热点。值得注意的是，2型糖尿病（T2DM）患者术后再狭窄的机率明显增高，但其机制还不清楚。最近我们通过凝胶缓释系统将肽基脯氨酰异构酶1(Pin1)抑制剂胡桃醌作用于T2DM大鼠股动脉损伤段的外膜，发现内膜增殖受抑制。另外，通过培养原代大鼠股动脉平滑肌细胞（VSMC），发现T2DM大鼠血清可明显抑制细胞凋亡，同时伴随Pin1上调。揭示Pin1可能是造成T2DM术后再狭窄的靶基因。本课题拟以慢病毒作载体，过表达或敲减VSMC内Pin1，观察细胞增值、凋亡和迁移变化，通过局部应用载shPin1慢病毒或胡桃醌在体内实验进一步阐明Pin1在T2DM再狭窄中的作用，并通过PPIase活性检测，免疫共沉淀，免疫印记等技术探讨其可能涉及的信号传导途径。该研究将为T2DM再狭窄的机制研究及防治提供有价值的线索。

中文关键词： 2型糖尿病；Pin1；pluronic F127；血管平滑肌细胞；再狭窄

英文摘要： Endovascular intervention has emerged in recent years and is considered as one of the most significant treatment for lower extremity atherosclerotic occlusive disease. How to reduce the restenosis rate is a medical research focus today. Of note, the risk of restenosis is particularly high in patients with type 2 diabetes mellitus (T2DM). However, the underlying molecular mechanism is unclear. Recently, we found that intima restenosis was inhibited by appling sustained-release system with Pin1 inhibitor juglone to the exposed adventitial surface of the femoral artery right after guidewire injury. In addition, the apoptosis rate of primary rat femoral artery smooth muscle cells (VSMC) cultured in T2DM rat serum was markedly inhibited, accompanied by upregulation of Pin1 level. These previous research suggest that Pin1 may play a key role in T2DM restenosis after endovascular treatment. On the basis of these significant studies, we try to observe the VSMC proliferation, apoptosis and migration changes by lentivirus mediated overexpression or knockdown of Pin1. Furthermore, the role of Pin1 in arterial restenosis in T2DM rats was elucidated by topical application of lentivirus mediated shPin1 or juglone in vivo experiments, and through PPIase activity assay, co-immunoprecipitation and immunoblotting et al, the possi

英文关键词： type 2 diabetes；Pin1；pluronic F127；vascular smooth muscle cells；restenosis