Van Beusecum Justin P, Xiao Liang, Barbaro Natalia R, Patrick David M, Kirabo Annet
Division of Clinical Pharmacology, Department of Medicine, Vanderbilt University Medical Center.
Division of Clinical Pharmacology, Department of Medicine, Vanderbilt University Medical Center;
J Vis Exp. 2019 Mar 5(145). doi: 10.3791/59124.
Excess dietary salt intake contributes to inflammation and plays a vital role in the development of hypertension. We previously found that antigen-presenting dendritic cells (DCs) can sense elevated extracellular sodium leading to the activation of the NADPH oxidase and formation of isolevuglandin (IsoLG)-protein adducts. These IsoLG-protein adducts react with self-proteins and promote an autoimmune-like state and hypertension. We have developed and optimized state-of-the-art methods to study DC function in hypertension. Here, we provide a detailed protocol for isolation, in vitro treatment with elevated sodium, and adoptive transfer of murine splenic CD11c cells into recipient mice to study their role in hypertension.
过量的膳食盐摄入会导致炎症，并在高血压的发展过程中起关键作用。我们之前发现，抗原呈递树突状细胞（DCs）能够感知细胞外钠升高，从而导致NADPH氧化酶激活并形成异前列腺素（IsoLG）-蛋白质加合物。这些IsoLG-蛋白质加合物会与自身蛋白质发生反应，促进类似自身免疫的状态和高血压。我们已经开发并优化了研究高血压中DC功能的先进方法。在此，我们提供一份详细的方案，用于分离小鼠脾脏CD11c细胞、用高钠进行体外处理以及将其过继转移到受体小鼠中，以研究它们在高血压中的作用。
