Tibetan medicine Wuwei Leze powder improves inflammatory mediated metabolic disorders and bone damage in collagen-induced arthritis rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Wuwei Leze Powder (WLP, སླྱེ་ཏྱེསལྔ་ཐང།) is a classic Traditional Tibetan medicine formula, which has certain clinical efficacy for rheumatoid arthritis (RA). Nevertheless, the pharmacological effects and potential therapeutic mechanisms of WLP on RA remain unclear.

AIM OF THE STUDY: The present study aimed to investigate the potential pharmacological mechanisms of anti- RA effect of WLP.

MATERIALS AND

METHODS

The chemical constituents of WLP were analyzed by UPLC-Q-TOF-MS. A collagen-induced arthritis (CIA) rat model was established to evaluate the swelling, arthritis index. Pathohistological staining and micro-CT were employed to evaluate the therapeutic effects of WLP. Subsequently, serum metabolomic analysis was conducted to elucidate the potential biomarkers and pathways. Finally, an enzyme-linked immunosorbent assay, along with immunofluorescence, RT-qPCR and western blotting, were utilized to verify the anti-RA mechanism of WLP.

RESULTS

A total of 109 chemical constituents from WLP were identified by UPLC-Q-TOF-MS. WLP reduced paw swelling, arthritis scores and organ index in the CIA rat model. Histopathological staining and micro-CT observed WLP possesses both anti-inflammatory and bone-protective properties. Subsequently, serum metabolomics identified 12 potential biomarkers, mainly related to amino acid metabolism and the mTOR signaling pathway. ELISA showed that WLP can regulate abnormal levels of inflammatory cytokines (IL-4, IL-10, TNF-α, IL-6, INF-γ and IL-17). Immunofluorescence demonstrated that WLP could regulate the expression of MMP-1, MMP-9, MMP-13 and RANKL. The effect of WLP on the expression of Wnt3a, Wnt10b, β-catenin, RANKL, OPG, p65 and MMP-9 were verified using RT-qPCR and WB, thereby elucidating the anti-arthritis mechanism of WLP.

CONCLUSION

The possible mechanism underlying the anti-RA of WLP involves the downregulation of the Wnt/RANKL/NF-κB axis, the restoration of abnormal host metabolite levels, the suppression of synovitis responses, and the attenuation of bone erosion. Considering the high variability of plant materials, the present study takes a batch of WLP as an example, which inevitably has limitations.