Polyphyllin VI screened from Chonglou by cell membrane immobilized chromatography relieves inflammatory pain by inhibiting inflammation and normalizing the expression of P2X3 purinoceptor

Abstract

Objective: Inflammatory pain is one of the most common diseases in daily life and clinic. In this work, we analysed bioactive components of the traditional Chinese medicine Chonglou and studied mechanisms of their analgesic effects.Material and methods: Molecular docking technology and U373 cells overexpressing P2X3 receptors combined with the cell membrane immobilized chromatography were used to screen possible CL bioactive molecules interacting with the P2X3 receptor. Moreover, we investigated the analgesic and anti-inflammatory effects of Polyphyllin VI (PPIV), in mice with chronic neuroinflammatory pain induced by CFA (complete Freund’s adjuvant).Results: The results of cell membrane immobilized chromatography and molecular docking showed that PPVI was one of the effective compounds of Chonglou. In mice with CFA-induced chronic neuroinflammatory pain, PPVI decreased the thermal paw withdrawal latency and mechanical paw withdrawal threshold and diminished foot edema. Additionally, in mice with CFA-induced chronic neuroinflammatory pain, PPIV reduced the expression of the pro-inflammatory factors IL-1, IL-6, TNF-α, and downregulated the expression of P2X3 receptors in the dorsal root ganglion and spinal cord.Conclusion: Our work identifies PPVI as a potential analgesic component in the Chonglou extract. We demonstrated that PPVI reduces pain by inhibiting inflammation and normalizing P2X3 receptor expression in the dorsal root ganglion and spinal cord.

Anti-inflammatory and analgesic properties of Polyphyllin VI revealed by network pharmacology and RNA sequencing

“…SRC, F2, ALB, and MAPK1 were found to be the core targets of the protein interaction network, while KEGG analysis was mainly concentrated highlighted in ammatory signaling pathways such as MAPK and P13K-Akt. This conclusion is consistent with the animal experiments showing that PP alleviates pain sensitization in the CFA-induced pain mice model by reducing the expression of in ammatory mediators [20] . The MAPK signaling pathway was also present in the subsequent KEGG enrichment analysis of DEGs of RNA-Seq.…”

“…The details of the CFA-induced in ammatory pain mice model and administration are presented in our previous article [20] .…”

“…The procession of protein extraction was described in detail [20] . 20 µg per sample of proteins were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and deposited onto a polyvinylidene uoride (PVDF) membrane after the concentration of proteins was determined.…”

“…The analgesic function of PP is arguably linked to the inhibition of P2X 3 purinoceptors alleviating in ammation [20] . In this study, we further clarify the analgesic and anti-in ammatory mechanism of PPIV.…”