基于IRE1/XBP1s通路介导的血管内皮细胞自噬-凋亡串话探讨补肾抗衰片干预动脉粥样硬化形成的分子机制

Atherosclerosis(AS)is a pathological basis for cardiovascular and cerebrovascular diseases. Vascular endothelial cell injury is the initiation factor of AS. Vascular endothelial cell apoptosis will initiate plaque formation, and endothelial cell autophagy has a dual role in protecting and damaging the arterial wall. Crosstalk of vascular endothelial cell autophagy and apoptosis affects the development of early stage of AS, and its regulatory mechanism is closely related to IRE1/XBP1s pathway. Early clinical studies have shown that Bushen Kangshuai Tablets treatment on AS disease is of efficacy certainly. Basic research confirmed that Bushen Kangshuai Tablets has the anti-apoptotic effect on vascular endothelial cell, but researches on effects of autophagy on endothelial cells was limited. This study is the extension of previous Project supported by National Natural Science Foundation of China. The crosstalk of vascular endothelial cells autophagy and apoptosis is used as the breakthrough point. Through the in vivo and in vitro experiments, Bushen Kangshuai Tablet are used to interfere. At the target of multi-link, detection of multi-level, intervention of multi-point, the study aimed to sequentially explore the molecular mechanism of its prevention and treatment of AS, so as to confirm the hypothesis that "Bushen Kangshuai Tablet can suppress the process of AS by regulating IRE1/XBP1s pathway and upregulating the expression of Beclin-1, thus inducing protective autophagy and inhibiting the apoptosis of vascular endothelial cells, which restoring vascular homeostasis and inhibiting of AS process", which further reveals action mechanism of the drug, and provide scientific basis for prevention and treatment of AS with traditional Chinese medicine.

动脉粥样硬化（AS）是心脑血管疾病的病理基础，血管内皮细胞损伤是AS的始动因素。血管内皮细胞凋亡会启动斑块形成，而内皮细胞自噬具有保护和损伤动脉壁的双重作用。血管内皮细胞自噬-凋亡串话影响AS早期病变，其调控机制与IRE1/XBP1s通路密切相关。前期临床研究表明，补肾抗衰片治疗AS性疾病疗效肯定，基础研究证实补肾抗衰片具有抗血管内皮细胞凋亡作用，但对内皮细胞自噬的影响缺乏深入探究。本课题系既往国自然项目基础的延伸，以血管内皮细胞自噬-凋亡串话为切入点，通过在体和离体实验，用补肾抗衰片进行干预，在靶点的多环节、检测的多水平、干预的多时点上，序贯探讨其防治AS的分子机制，以期证实“补肾抗衰片通过调控IRE1/XBP1s通路上调Beclin-1表达，诱导血管内皮细胞保护性自噬，抑制其过度凋亡，进而恢复血管稳态，阻抑AS进程”假说的真实性，进一步揭示该药物作用机理，为中医药防治AS提供科学依据。

动脉粥样硬化(AS)是心脑血管疾病的病理基础，血管内皮细胞损伤是AS的始动因素。在 AS发生早期斑块尚未完全成形之际，对受损内皮细胞进行干预以逆转或延缓本病的发展具有重要意义。血管内皮细胞凋亡会启动斑块形成，而内皮细胞自噬具有保护和损伤动脉壁的双重作用。血管内皮细胞自噬-凋亡串话影响AS早期病变，其调控机制与IRE1/XBP1s通路密切相关。前期临床研究表明，补肾抗衰片治疗AS性疾病疗效肯定，基础研究证实补肾抗衰片具有抗血管内皮细胞凋亡作用，但对内皮细胞自噬的影响缺乏深入探究。本项目以血管内皮细胞自噬-凋亡串话为切入点，通过在体和离体实验，用补肾抗衰片进行干预，在靶点的多环节、检测的多水平、干预的多时点上，序贯探讨其防治AS的分子机制。研究工作分为4个部分：补肾抗衰片对动脉粥样硬化模型小鼠动脉粥样硬化形成的影响、补肾抗衰片调控IRE1/XBP1s通路干预动脉粥样硬化形成的动态研究、同型半胱氨酸诱导血管内皮细胞损伤及内质网应激模型的建立、补肾抗衰片及其拆方调控IREI-XBP1通路干预血管内皮细胞自噬-凋亡串话的机制研究。在体实验部分采用高脂喂养方法建立ApoE-/-小鼠AS模型，从整体水平动态观察补肾抗衰片在不同时间点对AS形成的影响，离体实验部分采用不同浓度Hcy刺激人主动脉内皮细胞HAECs，建立主动脉内皮细胞损伤模型，从细胞水平观察血管内皮细胞损伤、凋亡和自噬形成情况，深入探究补肾抗衰片及其拆方抗Hcy诱导血管内皮细胞损伤的相关调控机制。本项目证实了补肾抗衰片通过调控IRE1/XBP1s通路，上调Beclin-1表达，诱导血管内皮细胞保护性自噬，抑制其过度凋亡，来起到保护血管内皮细胞，干预AS形成和发展的作用，为中医药防治AS提供了科学依据。