Cardiac ankyrin repeat protein attenuates cardiomyocyte apoptosis by upregulation of Bcl-2 expression
Cardiac ankyrin repeat protein attenuates cardiomyocyte apoptosis by upregulation of Bcl-2 expression
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心脏锚蛋白重复蛋白通过上调 Bcl-2 表达来减弱心肌细胞凋亡
DOI：
10.1016/j.bbamcr.2016.09.024
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发表时间：
2016-12-01
影响因子：
5.1
通讯作者：
Wang, Jian'an
中科院分区：
文献类型：
--
作者：
Zhang, Na;Ye, Feiming;Wang, Jian'an
心脏锚蛋白重复序列（CARP）是一种核转录辅助因子，作为肌小节的组成部分，在肌浆中具有额外的功能。先前的研究表明，CARP在心血管疾病中的表达增加，但其在心肌细胞凋亡中的作用尚不清楚且存在争议。在本研究中，我们探讨了CARP在缺氧/再氧化（H/R）诱导的心肌细胞凋亡中的可能作用及其潜在机制。分离新生小鼠心室心肌细胞，分别用编码Flag-tagged CARP （Ad-CARP）的腺病毒和编码CARP靶向shRNA （sh-CARP）的慢病毒感染。采用TUNEL染色和western blot分析裂解caspase-3对H/R条件下心肌细胞凋亡的影响。结果显示，H/ r诱导的Ad-CARP心肌细胞凋亡明显减少，sh-CARP心肌细胞凋亡明显增加，提示鲤鱼具有保护性抗凋亡作用。有趣的是，过表达的CARP主要分布在细胞核中，这与其调节转录活性的作用是一致的。qPCR分析显示，Bcl-2转录物在Ad-CARP心肌细胞中显著增加。ChIP和co-IP实验证实了CARP通过与转录因子GATA4的相互作用与Bcl-2启动子结合。综上所述，我们的研究结果表明，鲤鱼对H/R诱导的心肌细胞凋亡具有保护作用，可能是通过增加抗凋亡Bcl-2基因的表达。(C) 2016 Elsevier B.V.版权所有
Cardiac ankyrin repeat protein (CARP) is a nuclear transcriptional co-factor that has additional functions in the myoplasm as a component of the muscle sarcomere. Previous studies have demonstrated increased expression of CARP in cardiovascular diseases, however, its role in cardiomyocyte apoptosis is unclear and controversial. In the present study, we investigated possible roles of CARP in hypoxia/reoxygenation (H/R)-induced cardiomyocyte apoptosis and the underlying mechanisms. Neonatal mouse ventricular cardiomyocytes were isolated and infected with adenovirus encoding Flag-tagged CARP (Ad-CARP) and lentivirus encoding CARP targeted shRNA (sh-CARP), respectively. Cardiomyocyte apoptosis induced by exposure to H/R conditions was evaluated by TUNEL staining and western blot analysis of cleaved caspase-3. The results showed that H/R-induced apoptosis was significantly decreased in Ad-CARP cardiomyocytes and increased in sh-CARP cardiomyocytes, suggesting a protective anti-apoptosis role for CARP. Interestingly, over-expressed CARP was mainly distributed in the nucleus, consistent with its role in regulating transcriptional activity. qPCR analysis showed that Bcl-2 transcripts were significantly increased in Ad-CARP cardiomyocytes. ChIP and co-IP assays confirmed the binding of CARP to the Bcl-2 promoter through interaction with transcription factor GATA4. Collectively, our results suggest that CARP can protect against H/R induced cardiomyocyte apoptosis, possibly through increasing anti-apoptosis Bcl-2 gene expression. (C) 2016 Elsevier B.V. All rights reserved.