Siwei Zangmu Xiang alleviates rheumatoid arthritis via regulating RAGE signaling pathway: Integration of UPLC-Q-TOF-MS, network pharmacology, and in vivo/in vitro studies.

ETHNOPHARMACOLOGICAL RELEVANCE: Siwei Zangmu Xiang (SWZMX) Decoction powder is a classical Tibetan prescription used for the management of rheumatoid arthritis (RA). SWZMX has been widely used in clinical practice due to its potent anti-inflammatory, analgesic, and immunomodulatory properties. However, as a multi-component Tibetan medicine formula, its bioactive constituents and mechanisms of action in RA have not been fully elucidated.

AIM OF THE STUDY: Therefore, this study sought to reveal the active components in SWZMX and to explore its underlying mechanism of action on RA.

MATERIALS AND METHODS: Ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS) was used to profile the bioactive constituents of SWZMX. Network pharmacology was utilized to screen for RA-related molecular targets. The anti-inflammatory effects of SWZMX were then validated in vitro. The therapeutic efficacy of SWZMX was further evaluated in IL1RA-/- mice, a spontaneous arthritis model. UPLC-Q-TOF-MS was also used to identify SWZMX-derived metabolites in mouse blood, and network pharmacology was employed to predict the RA-associated targets of these metabolites. Finally, the RAGE signaling pathway implicated in SWZMX was verified using both cellular and animal experiments.

RESULTS: A total of 731 active constituents were identified in SWZMX. These constituents were predicted to act on RA primarily through the RAGE signaling pathway and exhibited significant anti-inflammatory activity in macrophages in vitro. Micro-CT analysis revealed that SWZMX markedly reduced ankle deformity and attenuated bone mineral density loss in IL1RA-/- mice. Histological examination showed that SWZMX significantly alleviated synovial inflammation and cartilage erosion. Twenty-eight anti-inflammatory or immunomodulatory metabolites were detected in the blood of SWZMX-treated mice, 17 of which were only present in the SWZMX administration group. Cellular and animal assays demonstrated that SWZMX suppressed the abnormal proliferation of inflammation-activated fibroblast-like synoviocytes (FLs) by inhibiting IL-6 and TNF-α secretion through the RAGE signaling pathway.

CONCLUSION: SWZMX aqueous extract alleviated synovial inflammation and bone damage in RA mice by suppressing the release of IL-6 and TNF-α through regulating the RAGE signaling pathway.