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基于脂质代谢诱导的炎症损伤的动脉粥样硬化进展中巨噬细胞分泌的 CD5L。

CD5L Secreted by Macrophage on Atherosclerosis Progression Based on Lipid Metabolism Induced Inflammatory Damage.

机构信息

Department of Cardiology, The First Affiliated Hospital of Nanchang University, Nanchang, 330006, Jiangxi, China.

出版信息

Arch Immunol Ther Exp (Warsz). 2022 Mar 6;70(1):10. doi: 10.1007/s00005-022-00643-y.

Abstract

To explore the molecular mechanism of exosomal protein CD5L secreted by macrophage to promote the progression of atherosclerosis. Twenty cases of patients with atherosclerosis (AS) and 20 cases of healthy subjects were collected. Morphological properties of exosomes were identified by transmission electron microscopy, and the marker proteins CD63 and CD81 of exosomes were measured by Western blot. The secretion of inflammatory factors in the blood supernatant were analyzed by ELISA. Atherosclerosis cell models were established by transwell and separated into three groups: first group was treated with exosome inhibitor (GW4869), second group was injected with CD5L protein and third group was model control. Morphological properties of exosomes were identified by transmission electron microscopy, and the marker proteins CD63 and CD81 of exosomes were measured by Western blot. The levels of TNF-α, IL-1β, IL-6, IL-13, IL-17A, IL-31 in the cells were analyzed by ELISA. Analysis of the expression and distribution of IL-17RA in vascular smooth muscle cells by immunofluorescence. The proteins of CD63, CD81, CD5L were high expressed in AS group compared to healthy subject group. Cell test results showed that protein levels of CD63, CD81, CD5L in AS group were much higher than that in normal group. Immunofluorescence showed that the expression level of IL-17RA in cell membrane was the highest in the AS model group, and the expression of IL-17RA was decreased in GW4869 group and CD5L group. Expression of inflammatory factors in AS was much higher than that in GW4869 group and CD5L group. The exosomal protein CD5L secreted by macrophage promotes the development of atherosclerosis based on lipid metabolism-induced inflammatory damage of vascular smooth muscle cells.

摘要

目的

探讨巨噬细胞来源的外泌体蛋白 CD5L 促进动脉粥样硬化(atherosclerosis,AS)进展的分子机制。

方法

收集 20 例 AS 患者和 20 例健康者,采用透射电镜鉴定外泌体形态特征,Western blot 检测外泌体标志蛋白 CD63、CD81,ELISA 法分析血上清中炎症因子的分泌。Transwell 建立 AS 细胞模型,分为外泌体抑制剂(GW4869)组、CD5L 蛋白组、模型对照组,Western blot 检测各组细胞 CD63、CD81、CD5L 蛋白的表达,ELISA 法分析细胞上清中 TNF-α、IL-1β、IL-6、IL-13、IL-17A、IL-31 水平,免疫荧光法分析血管平滑肌细胞中 IL-17RA 的表达和分布。

结果

与健康对照组比较,AS 组 CD63、CD81、CD5L 蛋白表达水平升高(P<0.05)。细胞实验结果显示,AS 组 CD63、CD81、CD5L 蛋白表达水平明显高于正常组(P<0.05)。免疫荧光结果显示,AS 模型组细胞膜上 IL-17RA 表达水平最高,GW4869 组和 CD5L 组 IL-17RA 表达减少,AS 组炎症因子表达水平明显高于 GW4869 组和 CD5L 组。

结论

巨噬细胞来源的外泌体蛋白 CD5L 通过脂质代谢诱导血管平滑肌细胞炎症损伤,促进 AS 的发生发展。

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