Nucleolin involved in myocardial ischaemic preconditioning via post-transcriptional control of HSPA1A expression
Nucleolin involved in myocardial ischaemic preconditioning via post-transcriptional control of HSPA1A expression
复制标题
核仁素通过 HSPA1A 表达的转录后控制参与心肌缺血预处理
DOI：
10.1093/cvr/cvu006
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发表时间：
2014-04-01
影响因子：
10.8
通讯作者：
Xiao, Xianzhong
中科院分区：
文献类型：
--
作者：
Jiang, Bimei;Liang, Pengfei;Xiao, Xianzhong
目的 最近的研究已经确定了核仁素在多种细胞过程中的关键作用，包括调节病毒复制和肿瘤形成。然而，核仁素在心肌预适应中的可能作用仍不清楚。 方法和结果我们使用了体内大鼠心肌缺血预适应（IP）模型（四个循环，5分钟缺血和10分钟再灌注）和细胞过氧化氢预适应（H2O2-PC）模型。我们发现，在大鼠心肌缺血预处理和新生大鼠心肌细胞H2O2-PC恢复过程中，核仁素mRNA和蛋白表达呈现时间依赖性增加。核仁素过度表达增强了 H2O2-PC 的保护作用，而核仁素消融则消除了 H2O2-PC 介导的心肌细胞保护作用。另一方面，核仁素过表达增加了心肌细胞中 HSPA1A mRNA 和 HSPA1A 蛋白表达的稳定性，而核仁素消除则消除了 H2O2-PC 诱导的心肌细胞中 HSPA1A 的上调。使用 RNA-蛋白质相互作用研究进一步鉴定了核仁素和 HSPA1A mRNA 之间的相互作用。报告基因检测依赖于 HSPA1A mRNA 的非翻译区 (UTR)，结果表明转录后调控主要归因于 3' UTR。最后，HSPA1A 反义寡核苷酸（ODN）减弱了核仁素对心肌细胞的保护作用。结论这些结果表明核仁素上调并通过 HSPA1A 表达的转录后控制参与缺血预处理的心肌保护。
Aims Recent studies have identified the critical roles of nucleolin in a variety of cellular processes, including regulation of viral replication and tumour formation. However, the possible roles of nucleolin in myocardial preconditioning remain undefined.Methods and results We used an in vivo rat myocardial ischaemic preconditioning (IP) model (four cycles of 5 min ischaemia and 10 min reperfusion) and cellular hydrogen peroxide preconditioning (H2O2-PC) models. We found that nucleolin mRNA and protein expression showed a time-dependent increase during the recovery of myocardial ischaemic preconditioning in rats and H2O2-PC in neonatal rat cardiomyocytes. Nucleolin overexpression enhanced the protective effects of H2O2-PC, whereas nucleolin ablation abrogated the H2O2-PC-mediated protection in cardiomyocytes. On the other hand, nucleolin overexpression increased the stabilization of the HSPA1A mRNA and the expression of HSPA1A protein in cardiomyocytes, whereas nucleolin ablation abrogated the up-regulation of HSPA1A induced by H2O2-PC in cardiomyocytes. An interaction between nucleolin and HSPA1A mRNA was further identified using the RNA-protein interaction studies. Reporter gene assays, which depended on the untranslated regions (UTR) of HSPA1A mRNA, revealed that the post-transcriptional regulation was mainly attributed to the 3' UTR. Finally, HSPA1A anti-sense oligonucleotides (as ODNs) attenuated the protective effect of nucleolin in cardiomyocytes.Conclusion These results indicate that nucleolin is up-regulated and involved in myocardial protection of ischaemic preconditioning via a post-transcriptional control of HSPA1A expression.