Study on the effect and mechanism of ZeXie decoction in treating MSU-induced acute gouty arthritis model through PI3K-AKT-mTOR signaling pathway

Abstract

Background: The incidence of acute gouty arthritis (AGA) is annually increasing, significantly detrimenting the quality of life for patients. ZeXie decoction (ZXT), composed of Atractylodes macrocephala Koidz and Alisma rhizome (Sam.), a timeless formula detailed in "Synopsis of the Golden Chamber" of Chinese medical sage Zhong-Jing Zhang, has shown promising clinical application in treating AGA. Alisol B, a principal component of ZXT, remains however, elusive in its mechanism of action against AGA. This study aimed to delve into the anti-inflammatory effects of Alisol B, a key component within ZXT, and to clarify its mechanism of action in the treatment of AGA.

Materials and methods: We adopted a network pharmacology approach to pinpoint the core targets and pathways involved in ZXT and Alisol B's treatment of AGA patients. Molecular docking was conducted using Autodock software to investigate potential interactions between Alisol B and its target proteins. An in vitro inflammation model was subsequently established. The impact of Alisol B on the expression of inflammatory factors in BMDMs treated with MSU was evaluated using RT-qPCR, supplemented by comparison with the PI3K agonist 740 Y-P (740YPDGFR) treated BMDMs. Subsequently, the expression of EGFR, PIK3CA, PIK3CB, and JAK2 - key players in the PI3K/AKT/mTOR signaling pathway - was assessed via RT-qPCR and Western blotting. Finally, the effect of MSU treatment and Alisol B's treatment on macrophage polarization was determined by flow cytometry.

Results: Findings from network pharmacology and molecular docking suggest that Alisol B may modulate the PI3K-AKT-mTOR signaling pathway to treat AGA. In vitro experiments revealed that Alisol B inhibited the expression of inflammatory vesicles and pro-inflammatory factors by suppressing MSU-induced activation of the PI3K/AKT/mTOR signaling pathway. Additionally, Alisol B improved the cellular inflammatory environment, fostering the production of M2 cells, which could potentially repair cells within the inflammatory environment.

Conclusion: Our research unveils that Alisol B curtails the production of inflammatory vesicles and pro-inflammatory cytokines while enhancing the production of anti-inflammatory factors by targeting the PI3K-AKT-mTOR signaling pathway in BMDMs. This may elucidate the pivotal mechanism of Alisol B in the treatment of AGA.

Keywords: AGA; Alisol B; Macrophage polarization; PI3K-AKT-mTOR signaling pathway; ZeXie decoction.