Abstract：
UNASSIGNED: Cellular communication among different types of vascular cells is indispensable for maintaining vascular homeostasis and preventing atherosclerosis. However, the biological mechanism involved in cellular communication among these cells and whether this biological mechanism can be used to treat atherosclerosis remain unknown. We hypothesized that endothelial autophagy mediates the cellular communication in vascular tissue through exosome-mediated delivery of atherosclerosis-related genes.
UNASSIGNED: Rapamycin and adeno-associated virus carrying Atg7 short hairpin RNA under the Tie (TEK receptor tyrosine kinase) promoter were used to activate and inhibit vascular endothelial autophagy in high-fat diet-fed ApoE-/- mice, respectively. miRNA microarray, in vivo and in vitro experiments, and human vascular tissue were used to explore the effects of endothelial autophagy on endothelial function and atherosclerosis and its molecular mechanisms. Quantitative polymerase chain reaction and miRNA sequencing were performed to determine changes in miRNA expression in exosomes. Immunofluorescence and exosome coculture experiments were conducted to examine the role of endothelial autophagy in regulating the communication between endothelial cells and smooth muscle cells (SMCs) via exosomal miRNA.
UNASSIGNED: Endothelial autophagy was inhibited in thoracic aortas of high-fat diet-fed ApoE-/- mice. Furthermore, rapamycin alleviated high-fat diet-induced atherosclerotic burden and endothelial dysfunction, while endothelial-specific Atg7 depletion aggravated the atherosclerotic burden. miRNA microarray, in vivo and in vitro experiments, and human vascular tissue analysis revealed that miR-204-5p was significantly increased in endothelial cells after high-fat diet exposure, which directly targeted Bcl2 to regulate endothelial cell apoptosis. Importantly, endothelial autophagy activation decreased excess miR-204-5p by loading miR-204-5p into multivesicular bodies and secreting it through exosomes. Moreover, exosomal miR-204-5p can effectively transport to SMCs, alleviating SMC calcification by regulating target proteins such as RUNX2.
UNASSIGNED: Our study revealed the exosomal pathway by which endothelial autophagy protects atherosclerosis: endothelial autophagy activation transfers miR-204-5p from endothelial cells to SMCs via exosomes, both preventing endothelial apoptosis and alleviating SMC calcification.
UNASSIGNED: URL: https://www.chictr.org.cn/; Unique identifier: ChiCTR2200064155.
UNASSIGNED: Rapamycin and adeno-associated virus carrying Atg7 short hairpin RNA under the Tie (TEK receptor tyrosine kinase) promoter were used to activate and inhibit vascular endothelial autophagy in high-fat diet-fed ApoE-/- mice, respectively. miRNA microarray, in vivo and in vitro experiments, and human vascular tissue were used to explore the effects of endothelial autophagy on endothelial function and atherosclerosis and its molecular mechanisms. Quantitative polymerase chain reaction and miRNA sequencing were performed to determine changes in miRNA expression in exosomes. Immunofluorescence and exosome coculture experiments were conducted to examine the role of endothelial autophagy in regulating the communication between endothelial cells and smooth muscle cells (SMCs) via exosomal miRNA.
UNASSIGNED: Endothelial autophagy was inhibited in thoracic aortas of high-fat diet-fed ApoE-/- mice. Furthermore, rapamycin alleviated high-fat diet-induced atherosclerotic burden and endothelial dysfunction, while endothelial-specific Atg7 depletion aggravated the atherosclerotic burden. miRNA microarray, in vivo and in vitro experiments, and human vascular tissue analysis revealed that miR-204-5p was significantly increased in endothelial cells after high-fat diet exposure, which directly targeted Bcl2 to regulate endothelial cell apoptosis. Importantly, endothelial autophagy activation decreased excess miR-204-5p by loading miR-204-5p into multivesicular bodies and secreting it through exosomes. Moreover, exosomal miR-204-5p can effectively transport to SMCs, alleviating SMC calcification by regulating target proteins such as RUNX2.
UNASSIGNED: Our study revealed the exosomal pathway by which endothelial autophagy protects atherosclerosis: endothelial autophagy activation transfers miR-204-5p from endothelial cells to SMCs via exosomes, both preventing endothelial apoptosis and alleviating SMC calcification.
UNASSIGNED: URL: https://www.chictr.org.cn/; Unique identifier: ChiCTR2200064155.
摘要：
■不同类型的血管细胞之间的细胞通讯对于维持血管稳态和预防动脉粥样硬化是必不可少的。然而,参与这些细胞间细胞通讯的生物学机制以及这种生物学机制是否可用于治疗动脉粥样硬化仍是未知的。我们假设内皮自噬通过外泌体介导的动脉粥样硬化相关基因的传递来介导血管组织中的细胞通讯。
■雷帕霉素和携带Tie（TEK受体酪氨酸激酶）启动子下的Atg7短发夹RNA的腺相关病毒用于激活和抑制高脂饮食喂养的ApoE-/-小鼠的血管内皮自噬，分别。miRNA微阵列,体内和体外实验，研究内皮自噬对内皮功能和动脉粥样硬化的影响及其分子机制。进行定量聚合酶链反应和miRNA测序以确定外来体中miRNA表达的变化。通过免疫荧光和外泌体共培养实验来研究内皮自噬在通过外泌体miRNA调节内皮细胞与平滑肌细胞（SMC）之间的通讯中的作用。
■在高脂饮食喂养的ApoE-/-小鼠的胸主动脉中，内皮自噬被抑制。此外，雷帕霉素减轻高脂饮食引起的动脉粥样硬化负担和内皮功能障碍，而内皮特异性Atg7耗竭加重了动脉粥样硬化负荷。miRNA微阵列,体内和体外实验，和人血管组织分析显示，miR-204-5p在高脂饮食暴露后在内皮细胞中显著增加，直接靶向Bcl2调节内皮细胞凋亡。重要的是,内皮自噬激活通过将miR-204-5p加载到多囊小体内并通过外泌体分泌来减少过量的miR-204-5p.此外，外泌体miR-204-5p能有效转运至SMCs,通过调节RUNX2等靶蛋白缓解SMC钙化。
■我们的研究揭示了内皮自噬保护动脉粥样硬化的外泌体途径：内皮自噬激活通过外泌体将miR-204-5p从内皮细胞转移到SMC，既防止内皮细胞凋亡，又减轻SMC钙化。
■URL：https://www。chictr.org.cn/;唯一标识符：ChiCTR2200064155。
■雷帕霉素和携带Tie（TEK受体酪氨酸激酶）启动子下的Atg7短发夹RNA的腺相关病毒用于激活和抑制高脂饮食喂养的ApoE-/-小鼠的血管内皮自噬，分别。miRNA微阵列,体内和体外实验，研究内皮自噬对内皮功能和动脉粥样硬化的影响及其分子机制。进行定量聚合酶链反应和miRNA测序以确定外来体中miRNA表达的变化。通过免疫荧光和外泌体共培养实验来研究内皮自噬在通过外泌体miRNA调节内皮细胞与平滑肌细胞（SMC）之间的通讯中的作用。
■在高脂饮食喂养的ApoE-/-小鼠的胸主动脉中，内皮自噬被抑制。此外，雷帕霉素减轻高脂饮食引起的动脉粥样硬化负担和内皮功能障碍，而内皮特异性Atg7耗竭加重了动脉粥样硬化负荷。miRNA微阵列,体内和体外实验，和人血管组织分析显示，miR-204-5p在高脂饮食暴露后在内皮细胞中显著增加，直接靶向Bcl2调节内皮细胞凋亡。重要的是,内皮自噬激活通过将miR-204-5p加载到多囊小体内并通过外泌体分泌来减少过量的miR-204-5p.此外，外泌体miR-204-5p能有效转运至SMCs,通过调节RUNX2等靶蛋白缓解SMC钙化。
■我们的研究揭示了内皮自噬保护动脉粥样硬化的外泌体途径：内皮自噬激活通过外泌体将miR-204-5p从内皮细胞转移到SMC，既防止内皮细胞凋亡，又减轻SMC钙化。
■URL：https://www。chictr.org.cn/;唯一标识符：ChiCTR2200064155。
