Uncovering the Mechanism of the Xingnaojing Injection against Ischemic Stroke Using a Combined Network Pharmacology Approach and Gut Microbiota Analysis

Abstract

Objective. To explore the brain protection mechanism of Xingnaojing injection (XNJ) against ischemic stroke (IS) by the network pharmacology approach and gut microbiota analysis. Methods. We used network pharmacology analysis to identify the active components of XNJ and its potential targets against IS and inflammatory bowel disease (IBD) and carried out network analysis, functional annotation, and pathway enrichment analysis. Then, transient middle cerebral artery occlusion (tMCAO) mice model was used to verify the molecular mechanism of XNJ. Results. 36 active compounds were identified from XNJ, and the effect of XNJ against IS was related to the VEGF signaling pathway, NF-kappa B signaling pathway, and gap junction. The effect of XNJ against IBD was related to the T cell receptor signaling pathway, NF-kappa B signaling pathway, and gap junction. In vitro experiments showed that XNJ significantly improved the neurological function of tMCAO mice, reduced the size of cerebral infarction, decreased the permeability of blood-brain barrier (BBB), downregulated the expressions of TLR4, MyD88, and NF-kappa B in the ischemic site, and upregulated the expressions of occludin and ZO-1 in the colon. High-throughput 16S rDNA gene sequencing showed that XNJ upregulated the levels of Akkermansia and downregulated the levels of Flavobacteriaceae, Deferribacteraceae, and Deferribacteres. XNJ increased the concentrations of the short-chain fatty acids (SCFAs) PA (propionate), VA (valerate), IBA (isobutyrate), and IVA (isovalerate) in the feces of the sham germ-free experiment group (SGFEG) mice. Conclusion. IS causes dysbiosis of some specific bacteria in the gut microbiota. XNJ is an effective treatment for IS, and its mechanism was related to improving intestinal barrier function and regulating intestinal flora and SCFAs. Network pharmacology revealed that XNJ acts through multiple targets and multiple pathways.

Role of gut microbiota in ischemic stroke: A narrative review of human and animal studies

“…Liu et al 63 Xingnaojing injection Improving intestinal barrier function and regulating intestinal flora and SCFAs.…”

“…We identified 140 articles, of which 32 articles were deemed irrelevant to our research topic, nine were reviews, two were letters to the editor, one was an editorial, one was a perspective piece, one was a comment, one was a duplicate publication, and one was an un‐peer‐reviewed preprint; these were excluded by two authors who independently assessed the articles. Ultimately, 92 studies, 5–96 including 38 human studies and 54 animal studies, were selected as being highly relevant to our research topic. The flowchart of the article selection process is depicted in Figure 1.…”

Chinese herbal injection for cardio-cerebrovascular disease: Overview and challenges

“…Salvia miltiorrhiza and safflower are currently widely used as clinical medicines to promote blood circulation and remove blood stasis . SMI was derived from Shendongyin in "Zhengyin Maizhi," written by Qin Jingming during the Ming Liu et al, 2022).…”