Objective To investigate the protective effect and mechanism of Buyang Huanwu Decoction (补阳还五汤, BYHWT) on intestinal mucosal barrier injury following cerebral ischemia-reperfusion injury (CIRI) in rats. Methods Network pharmacology was employed to screen potential targets of BYHWT for treating intestinal mucosal injury post-CIRI, followed by enrichment analysis. The improved Zea Longa suture method was used to prepare a rat CIRI model, which was randomly divided into model group and BYHWD low-, medium-, and high-dose (1.29, 2.57, 5.14 g/kg) groups, with six rats in each group, and six rats in sham group. After continuous administration for 5 d, the neurological deficit score and cerebral infarction area were measured. Hematoxylin-eosin (HE), TUNEL staining and electron microscopy were used to observe the pathological, apoptotic and ultrastructural changes of colon tissue. Immunohistochemistry and Western blotting were used to detect the protein expressions of Occludin and zonula occludin-1 (ZO-1) in colon tissue. Non-targeted metabolomics was performed on intestinal contents, and the results were integrated with network pharmacology to construct a “metabolite-reaction-enzyme-gene” network. Key pathways were validated using molecular docking, ELISA and Western blotting. Results Network pharmacology identified 22 core targets of BYHWT for treating post-CIRI intestinal mucosal injury, involving pathways such as lipid metabolism and mitogen activated protein kinase (MAPK). Compared with sham group, model group showed aggravated neurological damage (P < 0.001), increased cerebral infarction area (P < 0.05), disrupted colon mucosal structure, decreased expressions of Occludin and ZO-1 (P < 0.01), and increased cell apoptosis (P < 0.05). Compared with model group, BYHWT significantly improved the above indicators (P < 0.05, 0.01). Metabolomics discovered nine differential metabolite downregulation pathways, mainly enriched in steroid hormones, bile acids and arachidonic acid metabolism pathways. The integrated analysis of metabolomics and network pharmacology suggested that BYHWT exerted its effect by regulating arachidonic acid/dihydro-γ-linolenic acid prostaglandin metabolic pathway. Molecular docking showed that prostaglandin E₁ (PGE₁) bind well to prostaglandin endoperoxide synthase 2 (PTGS2). After BYHWT intervention, the level of PGE₁ in colon tissue was significantly increased (P < 0.05), while the expression of PTGS2 protein was significantly decreased (P < 0.05). Conclusion BYHWT may alleviate intestinal mucosal barrier injury following CIRI by modulating arachidonic acid/dihomo-γ-linolenic acid-PTGS2-prostaglandin metabolic network.

Buyang Huanwu Decoction  /  cerebral ischemia reperfusion injury  /  intestinal mucosa  /  metabolomics  /  network pharmacology  /  astragaloside IV  /  hydroxysafflor yellow A  /  ferulic acid  /  paeoniflorin  /  PTGS2