Department of Geriatrics, The First Affiliated Hospital of Fujian Medical University, Fuzhou, People's Republic of China.
Department of Cardiology, The First Affiliated Hospital of Gannan Medical University, Ganzhou, 341000, People's Republic of China.
Cell Commun Signal. 2021 Oct 11;19(1):103. doi: 10.1186/s12964-021-00779-y.
Transcription factor CREB is involved in the development of pulmonary hypertension (PH). However, little is known about the role and regulatory signaling of CREB in PH.
A series of techniques, including bioinformatics methods, western blot, cell proliferation and luciferase reporter assay were used to perform a comprehensive analysis of the role and regulation of CREB in proliferation of pulmonary artery smooth muscle cells (PASMCs) in PH.
Using bioinformatic analysis of the differentially expressed genes (DEGs) identified in the development of monocrotaline (MCT)- and hypoxia-induced PH, we found the overrepresentation of CRE-containing DEGs. Western blot analysis revealed a sustained increase in total- and phosphorylated-CREB in PASMCs isolated from rats treated with MCT. Similarly, an enhanced and prolonged serum-induced CREB phosphorylation was observed in hypoxia-pretreated PASMCs. The sustained CREB phosphorylation in PASMCs may be associated with multiple protein kinases phosphorylated CREB. Additionally, hierarchical clustering analysis showed reduced expression of the majority of CREB phosphatases in PH, including regulatory subunits of PP2A, Ppp2r2c and Ppp2r3a. Cell proliferation analysis showed increased PASMCs proliferation in MCT-induced PH, an effect relied on CREB-mediated transcriptional activity. Further analysis revealed the raised intracellular labile zinc possibly from ZIP12 was associated with reduced phosphatases, increased CREB-mediated transcriptional activity and PASMCs proliferation.
CREB pathway was overactivated in the development of PH and contributed to PASMCs proliferation, which was associated with multiple protein kinases and/or reduced CREB phosphatases and raised intracellular zinc. Thus, this study may provide a novel insight into the CREB pathway in the pathogenesis of PH. Video abstract.
转录因子 CREB 参与肺动脉高压（PH）的发生发展。然而，关于 CREB 在 PH 中的作用及其调控信号通路知之甚少。
我们采用了一系列技术，包括生物信息学方法、Western blot、细胞增殖实验和荧光素酶报告基因检测，对 CREB 在 PH 中肺动脉平滑肌细胞（PASMCs）增殖中的作用及其调控进行了全面分析。
利用生物信息学方法分析野百合碱（MCT）和低氧诱导的 PH 发展过程中差异表达基因（DEGs），我们发现 CRE 包含的 DEGs 明显过表达。Western blot 分析显示，MCT 处理的大鼠 PASMCs 中总 CREB 和磷酸化 CREB 持续增加。同样，在低氧预处理的 PASMCs 中也观察到增强和延长的血清诱导的 CREB 磷酸化。PASMCs 中持续的 CREB 磷酸化可能与磷酸化 CREB 的多种蛋白激酶有关。此外，层次聚类分析显示，PH 中大多数 CREB 磷酸酶的表达减少，包括 PP2A 的调节亚基、Ppp2r2c 和 Ppp2r3a。细胞增殖分析显示，MCT 诱导的 PH 中 PASMCs 增殖增加，该作用依赖于 CREB 介导的转录活性。进一步分析表明，升高的细胞内可动锌（可能来自 ZIP12）与磷酸酶减少、CREB 介导的转录活性增加和 PASMCs 增殖有关。
在 PH 的发生发展过程中，CREB 通路过度激活，促进 PASMCs 增殖，这与多种蛋白激酶和/或 CREB 磷酸酶减少以及细胞内锌升高有关。因此，本研究可能为 CREB 通路在 PH 发病机制中的作用提供新的见解。
