PUMA is critical for neonatal cardiomyocyte apoptosis induced by endoplasmic reticulum stress
PUMA is critical for neonatal cardiomyocyte apoptosis induced by endoplasmic reticulum stress
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DOI：
10.1016/j.cardiores.2006.10.001
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发表时间：
2007-01-01
影响因子：
10.8
通讯作者：
Erhardt, Peter
中科院分区：
文献类型：
--
作者：
Nickson, Philip;Toth, Ambrus;Erhardt, Peter
目的：Puma（p53-upregulated modulator of apoptosis）是Bcl-2蛋白家族中一个仅有促凋亡BH 3的成员，它与多种疾病的病理机制有关，包括癌症、AIDS和缺血性脑疾病。我们最近发现，Puma是小鼠心脏缺血/再灌注后心脏细胞死亡所必需的。由于缺血/再灌注也与内质网（ER）的压力，在本研究中，我们调查是否彪马有助于ER的压力依赖的组成部分的心肌细胞apoptosis.Methods：大鼠和小鼠的新生心肌细胞的原代培养物进行了处理3 μ M毒胡萝卜素或100毫微克/毫升（-1）衣霉素。通过腺病毒递送shRNA或靶向缺失puma基因来抑制Puma水平。RT-PCR和Western blotting检测Puma的表达。凋亡进行了评估，TUNEL法，caspase-3裂解，细胞色素c release.Results：我们已经表明，在大鼠新生心肌细胞，毒胡萝卜素或衣霉素治疗导致ER-应力，转录上调的彪马，和凋亡。最重要的是，如果通过腺病毒递送shRNA下调Puma表达或通过基因敲除小鼠中的靶向缺失消除Puma表达，则心肌细胞获得对ER应激诱导的凋亡的抗性。总之，我们的数据表明，Puma是ER应激诱导的心肌细胞凋亡的关键组分，抑制Puma活性可用于治疗心肌梗塞或通过阻断ER应激诱导的细胞凋亡来预防心力衰竭。(c)2006年欧洲心脏病学会。Elsevier B. V.出版，保留所有权利。
Objective: Puma (p53-upregulated modulator of apoptosis), a proapoptotic BH3-only member of the Bcl-2 protein family, has been implicated in the pathomechanism of several diseases, including cancer, AIDS, and ischemic brain disease. We have recently shown that Puma is required for cardiac cell death upon ischemia/reperfusion of mouse hearts. Since ischemia/reperfusion is also associated with endoplasmic reticulum (ER) stress, in the present study we investigated whether Puma contributes to the ER stress-dependent component of cardiomyocyte apoptosis.Methods: Primary cultures of rat and mouse neonatal cardiomyocytes were treated with 3 mu M thapsigargin or 100 ng mL(-1) tunicamycin. Puma levels were suppressed by adenoviral delivery of shRNA or targeted deletion of the puma gene. Puma expression was detected by RT-PCR and Western blotting. Apoptosis was assessed by TUNEL assay, caspase-3 cleavage, and cytochrome c release.Results: We have shown that in rat neonatal cardiac myocytes, thapsigargin or tunicamycin treatment led to ER-stress, transcriptional upregulation of Puma, and apoptosis. Most importantly, cardiac myocytes acquired resistance to ER stress-induced apoptosis if Puma expression was downregulated by adenoviral delivery of shRNA or eliminated by targeted deletion in knockout mice.Conclusion: Taken together, our data indicate that Puma is a critical component of ER stress-induced apoptosis in cardiac myocytes, and inhibition of Puma activity may be used to treat cardiac infarcts or prevent heart failure by blocking ER stress-induced apoptosis. (c) 2006 European Society of Cardiology. Published by Elsevier B.V. All rights reserved.