Hydrogen sulfide suppresses endoplasmic reticulum stress-induced endothelial-to-mesenchymal transition through Src pathway
Hydrogen sulfide suppresses endoplasmic reticulum stress-induced endothelial-to-mesenchymal transition through Src pathway
复制标题
硫化氢通过 Src 途径抑制内质网应激诱导的内皮间质转化
DOI：
10.1016/j.lfs.2015.11.025
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发表时间：
2016-01-01
期刊：
影响因子：
6.1
通讯作者：
Wang, Jing-Feng
中科院分区：
文献类型：
--
作者：
Ying, Ru;Wang, Xiao-Qiao;Wang, Jing-Feng
目的：在多种模型中，硫化氢（H₂S）通过抑制内质网（ER）应激来改善心脏纤维化。内皮 - 间充质转化（EndMT）与心脏纤维化的发展有关。因此，我们研究了H₂S是否能通过抑制内质网应激来减轻EndMT。
主要方法：在人脐静脉内皮细胞（HUVECs）中，衣霉素（TM）和毒胡萝卜素（TG）用于诱导内质网应激，4 - 苯基丁酸（4 - PBA）用于抑制内质网应激。通过蛋白质免疫印迹法、实时荧光定量聚合酶链反应（Real - Time PCR）和免疫荧光染色来检测内质网应激和EndMT。使用特异性抑制剂和小干扰RNA（siRNA）抑制Smad2和Src信号通路。通过透射电子显微镜进行超微结构检查。通过体外细胞迁移实验和管形成实验研究HUVECs的功能。
关键发现：在内质网应激条件下，内皮细胞标志物CD31的表达显著降低，而间充质标志物α - 平滑肌肌动蛋白（α - SMA）、波形蛋白和胶原蛋白1的表达增加，4 - PBA可抑制这种变化。此外，随着Smad2和Src激酶信号通路的激活，HUVECs转变为成纤维细胞样外观。抑制Src信号通路后，EndMT会受到显著抑制。TM降低了细胞裂解物中的H₂S水平，而H₂S预处理可保持内皮细胞的外观，降低内质网应激并改善内质网的扩张。H₂S还能下调间充质标志物的表达，上调内皮细胞标志物的表达，同时抑制Src信号通路。此外，H₂S部分恢复了HUVECs的迁移和管形成能力。
意义：这些结果表明，H₂S可通过Src信号通路抵御内质网应激诱导的EndMT，这可能是H₂S心脏保护作用的一种新机制。（C）2015爱思唯尔公司版权所有。
Aims: Hydrogen sulfide (H2S) ameliorates cardiac fibrosis in several models by suppressing endoplasmic reticulum (ER) stress. Endothelial-to-mesenchymal transition (EndMT) is implicated in the development of cardiac fibrosis. Therefore, we investigated whether H2S could attenuate EndMT by suppressing ER stress.Main methods: ER stress was induced by tunicamycin (TM) and thapsigargin (TG) and inhibited by 4-phenylbutyrate (4-PBA) in human umbilical vein endothelial cells (HUVECs). ER stress and EndMT were measured by Western blot, Real-Time PCR and immunofluorescence staining. Inhibition Smad2 and Src pathway were performed by specific inhibitors and siRNA. Ultrastructural examination was detected by transmission electron microscope. The functions of HUVECs were investigated by cell migration assay and tube formation in vitro.Key findings: Under ER stress, the expression of endothelial marker CD31 significantly decreased while mesenchymal markers alpha-SMA, vimentin and collagen 1 increased which could be inhibited by 4-PBA. Moreover, HUVECs changed into a fibroblast-like appearance with the activation of Smad2 and Src kinase pathway. After inhibiting Src pathway, EndMT would be significantly inhibited. TM reduced H2S levels in cell lysate and H2S pretreatment could preserve endothelial cell appearance with decreased ER stress and ameliorated dilation of ER. H2S could also downregulate the mesenchymal marker expression, and upregulate the endothelial markers expression, accompanied with the suppression of Src pathway. Moreover, H2S partially restored the capacity of migration and tube formation in HUVECs.Significance: These results revealed that H2S could protect against ER stress-induced EndMT through Src pathway, which may be a novel role for the cardioprotection of H2S. (C) 2015 Elsevier Inc. All rights reserved.