β1-adrenergic receptor autoantibodies from heart failure patients enhanced TNF-α secretion in RAW264.7 macrophages in a largely PKA-dependent fashion
β1-adrenergic receptor autoantibodies from heart failure patients enhanced TNF-α secretion in RAW264.7 macrophages in a largely PKA-dependent fashion
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DOI：
10.1002/jcb.24198
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发表时间：
2012-10-01
影响因子：
4
通讯作者：
Liu, Huirong
中科院分区：
文献类型：
--
作者：
Du, Yunhui;Yan, Li;Liu, Huirong
抗β 1-肾上腺素能受体（β 1-AA）细胞外第二环的自身抗体不仅有助于心力衰竭的易感性增加，而且通过与β 1-肾上腺素能受体结合而产生儿茶酚胺样作用，从而导致心肌重构。本研究旨在确定从心力衰竭患者血清中分离的β 1-AA是否会导致小鼠巨噬细胞样细胞系RAW264.7分泌TNF-a。研究人员采集了40名心力衰竭患者和40名健康受试者的血液样本。ELISA法检测β 1-AA滴度和TNF-a水平。采用CCK-8试剂盒和CFSE法检测β 1-AA对小鼠巨噬细胞样细胞系RAW264.7增殖的影响。Western blot法检测磷酸化vasp的表达。β 1-AA在心力衰竭患者中的出现频率高于健康人。心力衰竭患者分离的β 1-AA可促进TNF-a水平升高，可被选择性β 1-肾上腺素受体拮抗剂美托洛尔和β 1-肾上腺素受体细胞外第二环（β 1-AR-ECII）完全阻断，但仅被PKA抑制剂H89部分抑制。β 1-AA能促进RAW264.7细胞的体外增殖。同时，β 1-AA的存在显著增加了phospho-VASP的表达。这些结果首次表明，从心力衰竭患者分离的β 1-AA可以与RAW264.7细胞表面的β 1-AR结合，导致TNF-a的释放主要以pka依赖的方式进行。j .细胞。中国生物医学工程学报，2012,31(2):448 - 448。(c) 2012 Wiley期刊有限公司
Autoantibodies against the second extracellular loop of beta 1-adrenergic receptor (beta 1-AA) not only contribute to increased susceptibility to heart failure, but also play a causative role in myocardial remodeling through their catecholamine-like effects via binding with the beta 1-adrenergic receptor. The current study was designed to determine whether beta 1-AA isolated from the sera of heart failure patients could cause TNF-a secretion from the murine macrophage-like cell line RAW264.7. Blood samples were collected from 40 patients who had suffered heart failure, as well as from 40 healthy subjects. The titer of beta 1-AA and the level of TNF-a were detected using ELISA. The effect of beta 1-AA on murine macrophage-like cell line RAW264.7 proliferation was detected by CCK-8 kits and CFSE assay. Western blot assay was used to analyze the expression of phospho-VASP. beta 1-AA appeared more frequently in patients with heart failure than in healthy subjects. The beta 1-AA isolated from heart failure patients promoted an increase of TNF-a levels, which could be completely blocked by the selective beta 1-adrenergic receptor antagonist metoprolol and the second extracellular loop of beta 1-adrenergic receptor (beta 1-AR-ECII), but only partially inhibited by PKA inhibitor H89. Furthermore, the beta 1-AA could enhance the proliferation of RAW264.7 cells in vitro. Meanwhile, the expression of phospho-VASP was markedly increased in the presence of beta 1-AA. These results demonstrate for the first time that the beta 1-AA isolated from heart failure patients could bind with beta 1-AR on the surface of RAW264.7 cells, causing the release of TNF-a largely in a PKA-dependent fashion. J. Cell. Biochem. 113: 32183228, 2012. (c) 2012 Wiley Periodicals, Inc.