Analysis of necroptotic proteins in failing human hearts.
Analysis of necroptotic proteins in failing human hearts.
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DOI：
10.1186/s12967-017-1189-5
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发表时间：
2017-04-28
影响因子：
7.4
通讯作者：
Adameová A
中科院分区：
文献类型：
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作者：
Szobi A;Gonçalvesová E;Varga ZV;Leszek P;Kuśmierczyk M;Hulman M;Kyselovič J;Ferdinandy P;Adameová A
细胞损失和随后的收缩功能恶化是慢性心力衰竭（HF）的标志。虽然细胞凋亡作为心力衰竭进展的参与者进行了研究，但它不太可能解释非功能组织的总量。此外，有证据表明心力衰竭中存在坏死的心肌细胞。因此，本研究的目的是研究调节坏死性凋亡（程序性坏死的一种形式）的坏死性凋亡蛋白，从而评估其在人类终末期心力衰竭中的潜在作用。研究了健康对照 (C) 和因心肌梗塞 (CAD) 或扩张型心肌病 (DCM) 导致的终末期心力衰竭患者的左心室样本。对坏死性凋亡和凋亡标记物进行免疫印迹。分析 Triton X-114 分级样品以研究亚细胞定位的差异。与对照组相比，HF 组中观察到 RIP1（受体相互作用蛋白）、pSer227-RIP3 及其总水平的表达升高。另一方面，caspase-8（一种负向调节坏死性凋亡的促凋亡蛋白酶）表达下调，表明坏死性凋亡信号传导被激活。总混合谱系激酶结构域蛋白 (MLKL) 表达在各组之间没有差异；然而，所有 HF 样本中都存在 MLKL 的活性细胞毒性形式，而在对照中它们的表达水平几乎无法检测到。有趣的是，与 pSer358-MLKL 不同，pThr357-MLKL 在 DCM 中的表达高于 CAD。在 HF 中，RIP3 和 pThr357-MLKL 的亚细胞定位与坏死性凋亡信号传导的激活一致。主要凋亡标志物的表达并未表明细胞凋亡的重要性。这是第一个证据表明，CAD 或 DCM 病因的人类 HF 的坏死性凋亡标记物呈阳性，而坏死性凋亡标记物可能与 HF 的发生有关。本文的在线版本 (doi:10.1186/s12967-017-1189-5) 包含补充材料，可供授权用户使用。
Cell loss and subsequent deterioration of contractile function are hallmarks of chronic heart failure (HF). While apoptosis has been investigated as a participant in the progression of HF, it is unlikely that it accounts for the total amount of non-functional tissue. In addition, there is evidence for the presence of necrotic cardiomyocytes in HF. Therefore, the objective of this study was to investigate the necroptotic proteins regulating necroptosis, a form of programmed necrosis, and thereby assess its potential role in human end-stage HF. Left ventricular samples of healthy controls (C) and patients with end-stage HF due to myocardial infarction (CAD) or dilated cardiomyopathy (DCM) were studied. Immunoblotting for necroptotic and apoptotic markers was performed. Triton X-114 fractionated samples were analyzed to study differences in subcellular localization. Elevated expression of RIP1 (receptor-interacting protein), pSer227-RIP3 and its total levels were observed in HF groups compared to controls. On the other hand, caspase-8 expression, a proapoptotic protease negatively regulating necroptosis, was downregulated suggesting activation of necroptosis signaling. Total mixed-lineage kinase domain-like protein (MLKL) expression did not differ among the groups; however, active cytotoxic forms of MLKL were present in all HF samples while they were expressed at almost undetectable levels in controls. Interestingly, pThr357-MLKL unlike pSer358-MLKL, was higher in DCM than CAD. In HF, the subcellular localization of both RIP3 and pThr357-MLKL was consistent with activation of necroptosis signaling. Expression of main apoptotic markers has not indicated importance of apoptosis. This is the first evidence showing that human HF of CAD or DCM etiology is positive for markers of necroptosis which may be involved in the development of HF. The online version of this article (doi:10.1186/s12967-017-1189-5) contains supplementary material, which is available to authorized users.