Ethanol-Extracted Wenyujin Rhizoma Concisum Ameliorates Acetic Acid-Induced Pain and Improves Associated Central Inflammation by Inhibiting Microglial Inflammatory Activation

Objective This study investigated the analgesic effects and mechanisms of ethanol-extracted (e-WRC) versus water-extracted (w-WRC) Wenyujin Rhizoma Concisum in acetic acid-induced pain. Material and Methods Phytochemical analysis using UPLC-QTOF-MS. A mouse pain model was established via acetic acid injection. Writhing responses were assessed following administration of e-WRC or w-WRC (0.6, 3.6 g·kg -1 ). Cell viability was measured using CCK-8, while gene and protein expression were evaluated by qPCR, Western blot, or immunofluorescence. Transcriptional profiling of LPS-stimulated BV2 cells was conducted by RNA-seq. Results UPLC-QTOF-MS analysis confirmed that key anti-inflammatory compounds (furanodiene, curdione, germacrone, curcumin) were exclusively identified in e-WRC. e-WRC reduced writhing (78.67% vs. w-WRC 26.67%) and suppressed TNF-α and Iba-1 expression in brain tissue. In BV2 cells, e-WRC inhibited TNF-α and key inflammatory genes (Ikbke, Nfkbia, Nfkb1, Tlr2, Nlrp3), along with time-dependent downregulation of p-NF-κB. Conclusion e-WRC significantly alleviates visceral pain by inhibiting microglial activation and inflammation via the NF-κB and TLR signaling pathways.