Zhang Shuang, Wang Xueqian, Cheng Fafeng, Ma Chongyang, Fan Shuning, Xu Wenxiu, Jin Na, Liu Shuling, Lv Kai, Wang Qingguo
Beijing Key Laboratory, School of Basic Medical Sciences, Beijing University of Chinese Medicine, 11 Beisanhuandong Road, Chaoyang District, Beijing 100029, China.
School of Traditional Chinese Medicine, Capital Medical University, Beijing 100069, China.
Evid Based Complement Alternat Med. 2020 Aug 27;2020:2914579. doi: 10.1155/2020/2914579. eCollection 2020.
Ischemic stroke is the most common type of cerebrovascular accident worldwide. It causes long-term disability and death. Qingkailing (QKL) injection is a traditional Chinese patent medicine which has been clinically applied in the treatment of ischemic stroke for nearly thirty years. In the present study, network pharmacology combined with experimentation was used to elucidate the mechanisms of QKL. ADME screening and target prediction identified 62 active compounds and 275 targets for QKL. Topological screening of the protein-protein interaction (PPI) network was used to build a core PPI network consisting of 408 nodes and 17,830 edges. KEGG enrichment indicated that the main signaling pathway implicated in ischemic stroke involved hypoxia-inducible factor-1 (HIF-1). Experimentation showed that QKL alleviated neurological deficits, brain infraction, blood-brain barrier (BBB) leakage, and tight junction degeneration in a mouse ischemic stroke model. Two-photon laser scanning microscopy was used to evaluate BBB permeability and cerebral microvessel structure in living mice. HIF-1, matrix metalloproteinase-9 (MMP-9), and tight junction proteins such as occludin, zonula occludins-1 (ZO-1), claudin-5, and VE-Cadherin were measured by western blotting. QKL upregulated ZO-1 and downregulated HIF-1 and MMP-9. QKL has a multiapproach, multitarget, and synergistic effect against ischemic stroke.
缺血性中风是全球最常见的脑血管意外类型。它会导致长期残疾和死亡。清开灵（QKL）注射液是一种传统中成药，已在临床上用于治疗缺血性中风近三十年。在本研究中，采用网络药理学结合实验来阐明清开灵的作用机制。药物代谢动力学（ADME）筛选和靶点预测确定了清开灵的62种活性化合物和275个靶点。通过对蛋白质-蛋白质相互作用（PPI）网络进行拓扑筛选，构建了一个由408个节点和17830条边组成的核心PPI网络。京都基因与基因组百科全书（KEGG）富集分析表明，缺血性中风涉及的主要信号通路与缺氧诱导因子-1（HIF-1）有关。实验表明，清开灵可减轻小鼠缺血性中风模型的神经功能缺损、脑梗死、血脑屏障（BBB）渗漏和紧密连接退化。采用双光子激光扫描显微镜评估活体小鼠的血脑屏障通透性和脑微血管结构。通过蛋白质印迹法检测HIF-1、基质金属蛋白酶-9（MMP-9）以及紧密连接蛋白，如闭合蛋白、闭合蛋白-1（ZO-1）、claudin-5和血管内皮钙黏蛋白。清开灵上调ZO-1，下调HIF-1和MMP-9。清开灵对缺血性中风具有多途径、多靶点和协同作用。
