基于内质网应激调控血管内皮细胞凋亡研究补肾抗衰片干预动脉粥样硬化的分子机制

Atherosclerosis (AS) is the common pathologic basis of ischemic cardiovascular and cerebrovascular diseases. Causing the barrier of endothelial cells miopragia, apoptosis of vascular endothelial cell can initiate plaque formation and accelerate plaque development, plays an important role in the early stage of AS. Appropriate endoplasmic reticulum stress (ERS) conduces cell survival, while excess ERS can induce cell apoptosis, but the relationship between ERS and vascular endothelial cell apoptosis in the progress course of AS lacks deep research. Considering that the core pathogenesis of AS plaque formation is deficiency of spleen and kidney with phlegm stagnation, we use the traditional Chinese medicine Bushen Kangshuai Tablet to treat diseases with AS and receive good curative effect. Experiments show Bushen Kangshuai Tablet can attenuate AS through regulating the entire level of oxidative stress and inflammatory reaction. Based on the ground, this project further proposes a hypothesis that Bushen Kangshuai Tablet may reduce vascular endothelial cell apoptosis and inhibit AS development by regulating ERS. Using modern molecular biology technology, this research will study if Bushen Kangshuai Tablet can prevent apoptosis of vascular endothelial cell mediated by ERS at different time points in the progress course of AS and its molecular mechanism through both in vivo and in vitro experiments. From morphology, protein and gene aspects, we will investigate every link of vascular endothelial cell ERS apoptotic pathway and find possible effective targets of Bushen Kangshuai Tablet preventing and treating AS, also provide new thought for better control of AS from the angle of protecting arterial endothelial cells.

动脉粥样硬化（AS）是缺血性心脑血管疾病共同的病理基础，血管内皮细胞凋亡使内皮屏障功能减弱，启动斑块形成及加速斑块发展，在AS早期起重要作用。适当内质网应激（ERS）有助细胞生存，而过度ERS可诱导细胞凋亡，但ERS与AS进展过程中血管内皮细胞凋亡之间的关系缺乏深入研究。基于脾肾虚衰、痰浊停滞为AS形成核心病机的认识，前期运用补肾抗衰片治疗AS性疾病疗效肯定，实验表明可通过调节整体氧化应激和炎症反应而拮抗AS。本项目进一步提出该药还可能通过干预ERS调控血管内皮细胞凋亡，进而阻抑AS发展的假说。以在体、离体实验相结合的方法，运用现代分子生物学技术，从形态学、蛋白及基因水平，分析补肾抗衰片在AS进展过程中不同时间点对ERS介导血管内皮细胞凋亡的调控效应，并深入探索该药对血管内皮细胞ERS凋亡途径各环节干预的分子机制，从保护动脉内皮细胞角度阐明补肾抗衰片治疗AS原理，为临床防治AS提供新思路。

血管内皮细胞损伤及功能障碍是动脉粥样硬化（AS）最早期的事件及发病的始动环节，内皮细胞凋亡启动斑块形成及加速斑块发展，在AS早期起重要作用。适当内质网应激（ERS）有助细胞生存，而过度ERS可诱导细胞凋亡，但ERS与AS进展过程中血管内皮细胞凋亡之间的关系缺乏深入研究。本课题以ERS相关凋亡通路为切入点，在体、离体实验相结合，应用临床治疗AS性疾病效果确切、剂型稳定的补肾抗衰片作为干预药物，研究了补肾抗衰片调控血管内皮细胞ERS进而抑制其凋亡、拮抗AS的效应及机制。研究工作分为3个部分：补肾抗衰片调控ERS相关凋亡干预AS形成的研究、补肾抗衰片含药血清对血管内皮细胞凋亡影响的研究、补肾抗衰片单体调控IRE1-XBP1自噬途径干预血管内皮细胞凋亡的研究。先以高脂喂养建立的ApoE-/-小鼠AS模型为研究对象，从整体水平确定了补肾抗衰片在AS进展过程中不同时间点能够通过调控ERS减少细胞凋亡保护血管，且在AS早期更具有优势，对ERS激活的未折叠蛋白反应相关分子IRE1改善作用明显；继分别以ox-LDL、Hcy诱导的血管内皮细胞凋亡模型为研究对象，从细胞和分子层面对补肾抗衰片及其单体调控ERS减少细胞凋亡的效应及具体机制进行深入研究，明确了补肾抗衰片能够通过调控ERS介导的IRE1-XBP1-自噬-凋亡通路减少内皮细胞凋亡，从而发挥防治AS作用。研究紧扣中医“治未病”思想，从保护动脉内皮细胞角度阐明补肾抗衰片治疗AS原理，为理解AS形成机制和提出有效的干预措施提供线索，为中医药防治AS性疾病提供新思路。