Beyond the Database: Integration of Network Pharmacology and UHPLC-QTOF-MS/MS to Reveal the Neuroprotective Potential...

Network pharmacology is instrumental in understanding how TCM works by targeting specific pathways. This study examines different mechanisms using three network pharmacology approaches, focusing on the TCM "Bai Mi Decoction" (BMD). The neuroprotective effects of BMD were evaluated in a middle cerebral artery occlusion-reperfusion (MCAO/R) rat model. The composition of BMD extract (BMDE) and its components in brain (BMDB) were analyzed using UHPLC-QTOF-MS/MS, alongside documented constituents from the database (BMDD). A network pharmacological analysis was conducted to explore the similarities and differences in BMD's neuroprotective mechanisms. BMD showed a strong neuroprotective effect in MCAO/R rats, as indicated by lower neurological deficit scores, smaller cerebellar infarct sizes, and improved histopathological changes. Analysis identified 45, 11, and 22 components in BMDE, BMDB, and BMDD, respectively, with 29, 26, and 23 potential therapeutic targets. However, most database-listed compounds were not found in actual samples. Functional enrichment and pathway network analysis showed that BMDE and BMDB shared the most targets. MAPK1 was the only common target across all groups, targeted by crocetin, the sole shared compound. BMD proved highly effective in MCAO/R rats, with compounds found in BMD extracts or in vivo better reflecting the actual pharmacological mechanisms than database-derived ones.