Miao Fang, Lei Yangyang, Guo Yunfei, Ma Yongxia, Zhang Ye, Jia Binbin
Department of Prevention and Healthcare, Lanzhou Stomatology Hospital, Lanzhou, China.
Department of Cardiology, The Second People's Hospital of Lanzhou City, Lanzhou, China.
Cytojournal. 2024 Nov 15;21:42. doi: 10.25259/Cytojournal_76_2024. eCollection 2024.
Many different types of infectious oral diseases have been identified clinically, including chronic periodontitis. is the main pathogen causing chronic periodontitis, which is closely related to atherosclerosis (AS) and can promote the expression levels of caveolin 1 (Cav-1) and induced ribonucleic acid (RNA)-binding protein human antigen R (HuR). However, the roles of Cav-1 and its relationship with HuR in -mediated AS progression remain largely unknown. Here, we aimed to detect the role and molecular mechanisms of Cav-1 in -mediated AS.
To investigate the role of Cav-1 in -mediated AS, we infected human umbilical vein endothelial cells (HUVECs) with at a multiplicity of infection of 100:1 for 6, 12, and 24 h to simulate induced AS models and then transfected them with Cav-1 small interfering RNA to silence Cav-1. Combining molecular biology experimental techniques such as cell counting kit-8 assay, enzyme-linked immunosorbent assay, immunofluorescence staining, flow cytometry, Western blotting, and Oil Red O staining, and apolipoprotein E-deficient AS model mice, the impacts of Cav-1 on cell viability, inflammation, oxidative stress, apoptosis, Cav-1 and intercellular cell adhesion molecule-1 (ICAM-1) levels, and atherosclerotic plaque formation were investigated. Then, the relationship between Cav-1 and HuR was investigated through biotin pull-down and RNA immunoprecipitation assays, reverse transcription quantitative polymerase chain reaction, and Western blot.
can induce Cav-1 expression in a time- and dose-dependent manner ( < 0.05). This effect can inhibit the proliferation of HUVECs ( < 0.05). Cav-1 interference repressed inflammatory response, reactive oxygen species (ROS) and ICAM-1 levels, and apoptosis in the HUVECs ( < 0.05). Cav-1 messenger RNA was stabilized by HuR, which can bind to the 3' untranslated region of Cav-1. Increase in HuR level reversed the effects of Cav-1 silencing on ROS and ICAM-1 levels and apoptosis in the HUVECs ( < 0.05). In addition, the levels of inflammatory response, oxidative stress, and atherosclerotic plaque formation induced by in the mouse model were significantly reduced after Cav-1 expression was inhibited ( < 0.05).
HuR-activated Cav-1 may promote atherosclerotic plaque formation by modulating inflammatory response and oxidative stress, leading to AS.
临床上已鉴定出许多不同类型的感染性口腔疾病，包括慢性牙周炎。 是引起慢性牙周炎的主要病原体，其与动脉粥样硬化（AS）密切相关，并且可以促进小窝蛋白1（Cav-1）和诱导性核糖核酸（RNA）结合蛋白人抗原R（HuR）的表达水平。然而，Cav-1在介导的AS进展中的作用及其与HuR的关系在很大程度上仍然未知。在此，我们旨在检测Cav-1在介导的AS中的作用和分子机制。
为了研究Cav-1在介导的AS中的作用，我们以100:1的感染复数用 感染人脐静脉内皮细胞（HUVECs）6、12和24小时以模拟诱导的AS模型，然后用Cav-1小干扰RNA转染它们以沉默Cav-1。结合细胞计数试剂盒-8检测、酶联免疫吸附测定、免疫荧光染色、流式细胞术、蛋白质免疫印迹和油红O染色等分子生物学实验技术以及载脂蛋白E缺陷型AS模型小鼠，研究Cav-1对细胞活力、炎症、氧化应激、细胞凋亡、Cav-1和细胞间细胞黏附分子-1（ICAM-1）水平以及动脉粥样硬化斑块形成的影响。然后，通过生物素下拉和RNA免疫沉淀测定、逆转录定量聚合酶链反应和蛋白质免疫印迹研究Cav-1与HuR之间的关系。
可以以时间和剂量依赖性方式诱导Cav-1表达（<0.05）。这种作用可以抑制HUVECs的增殖（<0.05）。Cav-1干扰抑制了HUVECs中的炎症反应、活性氧（ROS）和ICAM-1水平以及细胞凋亡（<0.05）。HuR使Cav-1信使RNA稳定，其可以与Cav-1的3'非翻译区结合。HuR水平的增加逆转了Cav-1沉默对HUVECs中ROS和ICAM-1水平以及细胞凋亡的影响（<0.05）。此外，在抑制Cav-1表达后，小鼠模型中由 诱导的炎症反应、氧化应激和动脉粥样硬化斑块形成水平显著降低（<0.05）。
HuR激活的Cav-1可能通过调节炎症反应和氧化应激促进动脉粥样硬化斑块形成，导致AS。
