嗜铬粒蛋白 A（CGA）衍生多肽（CGA）通过 SOC 相关 Ca 信号抑制 TNF-α 诱导的血管内皮高通透性。
Chromogranin A (CGA)-derived polypeptide (CGA) inhibits TNF-α-induced vascular endothelial hyper-permeability through SOC-related Ca signaling.
机构信息
Department of Emergency, The First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, PR China; Department of Emergency, The Third People's Hospital of Chengdu, The Second Affiliated Chengdu Clinical College of Chongqing Medical University, Chengdu, Sichuan 610031, PR China.
Department of Emergency, The First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, PR China.
出版信息
Peptides. 2020 Sep;131:170297. doi: 10.1016/j.peptides.2020.170297. Epub 2020 May 5.
CGA (Vasostatin-1, VS-1) a N-terminal Chromogranin A (CGA)-derived peptide, has been shown to have a protective effect against TNF-α-induced impairment of endothelial cell integrity. However, the mechanisms of this effect have not yet been clarified. CGA (Chromofungin, CHR) is an important bioactive fragment of CGA. The present study aims to explore the protective effects of CHR on the vascular endothelial cell barrier response to TNF-α and its related Ca signaling mechanisms. EA.hy926 cells were used as a vascular endothelial culture model. The synthetic peptides CHR and CGA were assessed for their ability to suppress TNF-α-induced EA.hy926 cells hyper-permeability through Transwell® and TEER assays. Changes in [Ca] were measured through confocal laser scanning microscopy. SOC channel currents (Isoc) were measured via patch-clamp analysis. RT-PCR and western blot were used to analyze mRNA and protein expression of the transient receptor potential channels TRPC1 and TRPC4, respectively. FITC and rhodamine-phalloidin fluorescence were used to assess cell morphology and the distribution of MyPT-1 and F-actin. Compared to untreated cells, TNF-α increased the permeability of EA.hy926 cells that was inhibited by pre-treatment with CHR (10-1000 nM) in concentration-dependent manner, and the effect was most obvious at 100 nM, but CGA (100 nM) had no effect. TNF-α treatment increased the phosphorylation of MyPT-1 and stress fiber formation. CHR (10-1000 nM) pretreatment inhibited the cytoskeletal rearrangements and increased [Ca] in response to TNF-α treatment. CHR also reduced TRPC1 expression following TNF-α induction. Similar to SOC inhibitor 2-APB, CHR suppressed IP mediated SOC activation. These findings suggest that CHR inhibits TNF-α-induced Ca influx and protects the barrier function of vascular endothelial cells, and that these effects are related to the inhibition of SOC and Ca signaling by CHR.
CGA（Vasostatin-1，VS-1）是一种源自 N 端嗜铬粒蛋白 A（CGA）的肽，已被证明对 TNF-α 诱导的内皮细胞完整性损伤具有保护作用。然而，其作用机制尚未阐明。CGA（Chromofungin，CHR）是 CGA 的重要生物活性片段。本研究旨在探讨 CHR 对 TNF-α 诱导的血管内皮细胞屏障反应的保护作用及其相关的 Ca 信号机制。EA.hy926 细胞被用作血管内皮培养模型。通过 Transwell®和 TEER 测定评估合成肽 CHR 和 CGA 抑制 TNF-α诱导的 EA.hy926 细胞高通透性的能力。通过共聚焦激光扫描显微镜测量 [Ca]的变化。通过膜片钳分析测量 SOC 通道电流（Isoc）。使用 RT-PCR 和 Western blot 分别分析瞬时受体电位通道 TRPC1 和 TRPC4 的 mRNA 和蛋白表达。使用 FITC 和 rhodamine-phalloidin 荧光评估细胞形态以及 MyPT-1 和 F-肌动蛋白的分布。与未经处理的细胞相比，TNF-α增加了 EA.hy926 细胞的通透性，这种通透性可被 CHR（10-1000 nM）预处理以浓度依赖性方式抑制，并且在 100 nM 时效果最明显，但 CGA（100 nM）没有作用。TNF-α处理增加了 MyPT-1 的磷酸化和应力纤维形成。CHR（10-1000 nM）预处理抑制了细胞骨架重排，并增加了对 TNF-α处理的 [Ca]。CHR 还降低了 TNF-α诱导后的 TRPC1 表达。与 SOC 抑制剂 2-APB 相似，CHR 抑制了 IP 介导的 SOC 激活。这些发现表明，CHR 抑制 TNF-α诱导的 Ca 内流并保护血管内皮细胞的屏障功能，并且这些作用与 CHR 抑制 SOC 和 Ca 信号有关。
Zotero 插件