Department of Neurology, Shanghai Jiao Tong University School of Medicine Affiliated Renji Hospital, Shanghai, China.
Department of Neurology, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, China.
Stroke Vasc Neurol. 2021 Jun;6(2):244-251. doi: 10.1136/svn-2020-000584. Epub 2020 Nov 27.
Hydrogen sulphide (HS) is considered as the third member of the gasotransmitter family, along with nitric oxide (NO) and carbon monoxide. HS has been reported to induce angiogenesis by promoting the growth, migration and tube-like structure formation of endothelial cells. Those studies were conducted in conditions of cell culture, mouse Matrigel plug assay model, rat wound healing model or rat hindlimb ischaemia model. Recent in vivo studies showed the physiological importance of HS in muscle angiogenesis. However, the importance of endogenous HS for brain angiogenesis during development remains unknown. We therefore aimed at determining the role of HS in brain vascular development.
Both knockdown and knockout of HS-producing enzymes, cystathionine β-synthase () and cystathionine γ-lyase (), using morpholino oligonucleotides and clustered regularly interspaced short palindromic repeats/Cas9-mediated mutation, impaired brain vascular development of larval zebrafish. Incubation with the slow-releasing HS donor GYY4137 alleviated the defects of brain vascular development in and morphants. Quantitative analysis of the midbrain vascular network showed that HS enhances angiogenesis without affecting the topological structure of the brain vasculature. Mechanically, nitric oxide synthase 2a () expression and NO production were decreased in both and morphants. Overexpression of by coinjection of or MO with full-length zebrafish nos2a mRNA alleviated the brain vascular developmental defects in and morphants.
We conclude that HS promotes brain developmental angiogenesis via the NOS/NO pathway in zebrafish.
硫化氢（HS）被认为是气体递质家族的第三个成员，与一氧化氮（NO）和一氧化碳一起。有报道称 HS 通过促进内皮细胞的生长、迁移和管状结构形成来诱导血管生成。这些研究是在细胞培养、小鼠 Matrigel plugs 模型、大鼠伤口愈合模型或大鼠后肢缺血模型中进行的。最近的体内研究表明 HS 在肌肉血管生成中的生理重要性。然而，内源性 HS 在发育过程中对大脑血管生成的重要性尚不清楚。因此，我们旨在确定 HS 在大脑血管发育中的作用。
使用 morpholino 寡核苷酸和聚类规则间隔短回文重复序列/Cas9 介导的突变敲低和敲除 HS 产生酶胱硫醚 β-合酶（CBS）和胱硫醚 γ-裂合酶（CSE），损害了幼鱼斑马鱼的大脑血管发育。用缓慢释放的 HS 供体 GYY4137 孵育可缓解 和 突变体的大脑血管发育缺陷。中脑血管网络的定量分析表明，HS 增强了血管生成，而不影响大脑脉管系统的拓扑结构。在机制上，NOS2a 的表达和 NO 的产生在 和 突变体中均降低。通过将全长 zebrafish nos2a mRNA 与 或 MO 共注射，过表达 可减轻 和 突变体的大脑血管发育缺陷。
我们的结论是，HS 通过 NOS/NO 通路促进斑马鱼大脑发育性血管生成。
