Exploration of the TNF-α signaling pathway in cerebral ischemia-reperfusion injury and its modulation by Gui Zhi Fu Ling Wan: Insights from network pharmacology

Objective The current study endeavors to elucidate the precise role and underlying molecular mechanism of GZFL in cerebral ischemia-reperfusion injury (CIRI). Methods Public databases for network pharmacology were used to identify the compound ingredients of GZFL, their associated targets, and the protein targets related to CIRI. Utilizing the Autodock software, a molecular docking analysis was conducted to model and simulate the intricate interactions occurring between the pivotal compounds of GZFL and the central protein implicated in CIRI. Subsequently in vivo and in vitro experiments were conducted to investigate the impact of GZFL on CIRI. Results Network pharmacology analysis revealed that there were 115 common targets shared between GZFL and CIRI. Molecular docking experiments demonstrated a significant binding activity between TNF and two compounds of GZFL, namely Quercetin and beta-sitosterol. In vitro studies exhibited that GZFL effectively inhibited neuronal cell apoptosis induced by OGD/R, improved cell viability, and reduced the levels of inflammatory factors IL-1β, IL-6, and MCP-1 following OGD/R treatment. Additionally, GZFL treatment significantly decreased the expression of MDA induced by OGD/R, while upregulating the expression of SOD and GSH-Px. Mechanistic investigations illustrated that GZFL treatment was able to suppress the expression of TNF-α induced by OGD/R. In vivo studies demonstrated that GZFL treatment ameliorated neurological function scores and pathological alterations in rat brain tissue. Conclusion This research suggests that GZFL may hold promise as a therapeutic drug for CIRI subsequent to an ischemic stroke. Its protective effects are likely primarily mediated through the TNF-α pathway.