Decoding Gujin Luyan Xuming decoction: How ancient wisdom meets modern science in promoting angiogenesis to ameliorate cerebral infarction.

Ethnopharmacological relevance: Gujin Luyan Xuming decoction (XMD), a traditional chinese medicine formula documented in the "Synopsis of the Golden Chamber", is an effective prescription for treating cerebral infarction with notable clinical efficacy. However, its molecular mechanism remains to be fully elucidated.

Aim of the study: This research seeks to clarify the molecular mechanisms by which XMD facilitates angiogenesis following cerebral infarction.

Methods: The therapeutic levels of XMD on cerebral infarction were assessed in vivo, and the bioactive constituents of XMD were identified in systemic circulation. Afterward, the RNA-seq dataset GSE137482 and single-cell sequencing dataset GSE225948 were analyzed using R software, in conjunction with WGCNA, predicted drug targets derived from bioactive components, and angiogenesis-related gene sets, to identify key drug targets. We also performed molecular docking and molecular dynamics simulation. Finally, both in vivo and in vitro experiments were conducted for validation.

Results: Our research showed that XMD-H yielded the most significant effect on improving ischemic brain injury. A total of 33 blood-entering components of XMD were identified using HPLC-MS/MS analysis. Network pharmacology, RNA sequencing, single-cell sequencing, and further investigations identified RAB11A as the primary target of XMD's effect in cerebral infarction. According to molecular docking and dynamics simulation, the bioactive components of XMD could partially bind with the RAB11A protein. Animal experiments demonstrated that XMD could enhance the expression of RAB11A, Wnt5a, β-catenin, VEGFA, VEGFR2, and other proteins. The outcomes of in vitro experiments indicated that XMD could further augment cell migration, lumen formation, and the expression of the aforementioned proteins. The silencing of RAB11A led to a partial reduction in XMD's efficacy in promoting cell migration, lumen formation, and the expression of proteins such as RAB11A, Wnt5a, and VEGFR2.

Conclusions: XMD could affect the Wnt5a/β-catenin signaling pathway through RAB11A, thereby enhancing angiogenesis in cerebral infarction and mitigating cerebral ischemia injury.