Zhang Mingming, Yue Xiangyong, Zhao Xueping, Lu Yonggang, Liu Hongtao, Zhang Zhe, Ma Huan, Wang Xing, Xing Hanying
Clinical Medicine Research Center, Hebei General Hospital, Shijiazhuang, Hebei 050051, China.
Department of Oncology, Hebei General Hospital, Shijiazhuang, Hebei 050051, China.
Aging (Albany NY). 2023 Dec 26;15(24):15196-15212. doi: 10.18632/aging.205342.
Notch-1 signaling is significantly associated with the occurrence and development of atherosclerosis (AS). However, the molecular mechanisms underlying the specific deletion of Notch-1 in AS-associated macrophages are not fully understood. This study aimed to investigate the effects of Notch-1 in AS.
Tissue samples were obtained from atherosclerotic segments of human carotid arteries. Immunofluorescence staining showed that Notch-1 was significantly colocalized with macrophages (CD68+), and Notch-1 staining was increased in human vulnerable plaques. Notch-1/ApoE mice were generated in which Notch-1 was selectively inactivated in macrophages, and WT for littermate control mice (ApoE/Notch-1). A control group was then established. All mice fed with a high-fat and Oil Red O, Movat, a-SMA, CD68, and Sirius red staining were used to evaluate the morphology. Specific deletion of Notch-1 in macrophages repressed the pathophysiology of AS. Immunofluorescent staining and Western blotting revealed that Notch-1 repressed M1 and M2 responses in AS. Here, GSEA revealed that Notch-1 activation and PI3K signaling were statistically significantly correlated with each other, and Notch-1 was involved in the regulation of the PI3K signaling pathway. In the experiments, the secretion of Arg-1 and exosomes was classified by peritoneal macrophages of Notch-1/ApoE and Notch-1/ApoE mice. Immunohistochemistry staining and Western blotting were used to measure the expression levels of Notch1, PI3K, p-PI3K, AKT, p-AKT, Arg-1, IL-6, CD36, SREBP-1, CD206, iNOS, cleaved-caspase-3/-9, Bax, CD9, Alix and TSG101 in the peritoneal macrophages and exosomes, respectively.
The specific deletion of Notch-1 in macrophage represses the formation and development of AS via the PI3K/AKT signaling pathway.
Notch-1信号通路与动脉粥样硬化（AS）的发生发展密切相关。然而，AS相关巨噬细胞中Notch-1特异性缺失的分子机制尚未完全明确。本研究旨在探讨Notch-1在AS中的作用。
从人类颈动脉粥样硬化节段获取组织样本。免疫荧光染色显示，Notch-1与巨噬细胞（CD68+）显著共定位，且在人类易损斑块中Notch-1染色增加。构建了巨噬细胞中Notch-1选择性失活的Notch-1/ApoE小鼠，同窝对照小鼠（ApoE/Notch-1）为野生型。然后设立对照组。所有小鼠均喂食高脂饲料，采用油红O、Movat、α-SMA、CD68和天狼星红染色评估形态。巨噬细胞中Notch-1的特异性缺失抑制了AS的病理生理过程。免疫荧光染色和蛋白质印迹法显示，Notch-1抑制AS中的M1和M2反应。在此，基因集富集分析（GSEA）显示，Notch-1激活与PI3K信号通路在统计学上显著相关，且Notch-1参与PI3K信号通路的调控。在实验中，对Notch-1/ApoE和Notch-1/ApoE小鼠的腹腔巨噬细胞分泌的精氨酸酶-1（Arg-1）和外泌体进行分类。分别采用免疫组织化学染色和蛋白质印迹法检测腹腔巨噬细胞和外泌体中Notch1、PI3K、p-PI3K、AKT、p-AKT、Arg-1、白细胞介素-6（IL-6）、CD36、固醇调节元件结合蛋白-1（SREBP-1）、CD206、诱导型一氧化氮合酶（iNOS）、裂解的半胱天冬酶-3/-9、Bax、CD9、Alix和TSG101的表达水平。
巨噬细胞中Notch-1的特异性缺失通过PI3K/AKT信号通路抑制AS的形成和发展。
