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目的:动脉粥样硬化(atherosclerosis,AS)是一种慢性炎症过程,其发病机制复杂,目前公认的是内皮损伤反应学说。有研究称,细胞骨架蛋白与内皮细胞功能及内皮相关信号通路密切相关。Testin是一种新发现的细胞骨架蛋白,主要分布于黏着斑和细胞间连接的区域,参与细胞间的黏附,调节细胞的运动。然而,Testin在AS中的作用尚不明确。方法:利用家兔高脂饮食喂养16周建立冠脉AS模型,分离冠脉,免疫荧光染色观察Testin的分布,检测AS兔与正常兔Testin的表达水平;分离培养人脐静脉内皮细胞(HUVECs),分别予以1 mg/L LPS、40 mg/L ox-LDL刺激6 h,检测Testin表达水平的变化;于HUVECs过表达或下调表达Testin,观察其对细胞增殖、凋亡、迁移、招募单核细胞能力及AS相关因子表达的影响。结果:染色示Testin主要表达于血管内皮层并与内皮标志物CD31存在共定位,AS兔Testin m RNA和蛋白表达水平较正常兔降低(P<0.05);给予HUVECs刺激后,Testin m RNA和蛋白表达水平降低(P<0.05);过表达或下调Testin,不影响细胞增殖和凋亡能力(P>0.05);过表达Testin可促进HUVECs迁移而抑制单核细胞与其黏附,下调Testin则呈现相反作用(P<0.05);过表达Testin可升高一氧化氮合酶水平而降低单核细胞趋化因子1、基质金属蛋白酶2水平,下调Testin则呈现相反趋势(P<0.05)。结论:Testin可通过影响内皮功能发挥抗AS作用。
AIM: Atherosclerosis (AS) is a chronic inflammatory process with complicated pathogenesis, and is currently recognized as the theory of endothelial injury response. Some studies have shown that cytoskeletal proteins and endothelial cell function and endothelial-related signal pathways are closely related. Testin is a newly discovered cytoskeletal protein, mainly located in the focal adhesion and intercellular junction area, involved in the adhesion between cells, regulating cell movement. However, the role of Testin in AS is not clear. Methods: The coronary arteries were isolated from coronary arteries by high-fat diet for 16 weeks in rabbits. The distribution of Testin was detected by immunofluorescence staining. The expression of Testin in AS rabbits and normal rabbits was detected. Human umbilical vein endothelial cells (HUVECs) , Respectively, to be 1 mg / L LPS, 40 mg / L ox-LDL stimulation 6 h, Testin expression levels were detected; HUVECs overexpression or down-regulated expression of Testin, observed the proliferation, apoptosis, migration, Cell competence and expression of AS-related factors. Results: The expression of Testin mRNA was mainly localized in the vascular endothelium and co-localized with endothelial marker CD31. The expression of Testin mRNA and protein in AS rabbits was lower than that in normal rabbits (P <0.05). After stimulation with HUVECs, (P <0.05). Over-expression or down-regulation of Testin did not affect cell proliferation and apoptosis (P> 0.05). Overexpression of Testin could promote the migration of HUVECs and inhibit the adhesion of monocytes to Testin, (P <0.05). Overexpression of Testin increased the level of nitric oxide synthase and decreased the levels of MCP-1 and MMP-2. The down-regulation of Testin showed the opposite trend (P <0.05). Conclusion: Testin exerts anti-AS effects by affecting endothelial function.