Combination of Withania coagulans and Fagonia cretica ameliorates hyperuricemia by re-modulating gut microbiota-derived spermidine and traumatic acid.

BACKGROUND: Hyperuricemia (HU), characterized by elevated serum uric acid (UA) levels, is associated with metabolic disorders, renal diseases, gout and gut dysbiosis. Withania coagulans (WC) and Fagonia cretica (FC) are medicinal plants traditionally used in Pakistan for their antioxidant and anti-inflammatory properties; however, their potential in HU management remains unexplored.

PURPOSE: This study investigates the efficacy of WC and FC crude extracts in ameliorating HU and explores the underlying mechanisms in mice models with HU.

METHODS: Liquid chromatography and mass spectrometry (LC-MS) were adopted to characterize the major components within the crude extracts of WC and FC. HU was induced in BALB/c mice using potassium oxonate (PO) and adenine (A) for 21 days. Mice models were treated orally with WC (200mg/kg) and FC (200mg/kg) crude extracts during HU induction. Serum UA, creatinine, and inflammatory markers were analyzed using ELISA. The molecular expression of UA transporters (ABCG2 and SLC2A9) and TLR4/MyD88/NF-κB pathway components were assessed via qRT-PCR and immunoblotting. Gut microbiota composition was analyzed via fecal 16S rRNA sequencing, and metabolomic profiling identified key metabolites. In vitro studies were conducted using Hep G2 and HT-29 cells to examine the effects of gut microbiota-derived metabolites on xanthine oxidase (XOD) activity and UA transporters.

RESULTS: Treatment with WC and FC markedly lowered serum UA, creatinine, and hepatic inflammatory cytokine levels in HU mice. WC showed strong effects in inhibiting XOD activity, while FC significantly upregulated intestinal UA transporters, suggesting their different mechanisms of action. WC and FC treatment reduced pro-inflammatory cytokines (IL-1β, TNF-α, IL-6, IL-18) and LPS levels via downregulating the TLR4/MyD88/NF-κB pathway, indicating a strong anti-inflammatory effect. Additionally, the anti-inflammatory cytokine IL-10 was markedly increased. WC extracts restored gut microbial diversity by increasing the levels of Bifidobacterium, while FC increased Eubacterium xylanophilum. Metabolomic analysis of the gut microbiota revealed increased spermidine (SP) levels in WC-treated mice and traumatic acid (TA) levels in FC-treated mice, which contributed to reduced UA synthesis and enhanced excretion. In vitro cell-line and transcriptomic analyses demonstrated that key metabolites, SP (showing XOD inhibition) and TA (enhancing UA excretion), alleviated liver and intestinal dysfunction by suppressing TNF and IL-17 signaling pathways. The combined treatment exhibited synergistic effects.

CONCLUSION: This study highlights the complementary mechanisms of Withania coagulans (WC) and Fagonia cretica (FC) in managing hyperuricemia (HU). WC primarily inhibits xanthine oxidase (XOD), reducing uric acid (UA) synthesis, while FC enhances UA excretion by upregulating transporters ABCG2 and SLC2A9. The WC extract alleviated PO+A-induced HU through its anti-inflammatory properties and modulation of gut microbiota-derived SP production, leading to XOD inhibition by suppressing the TNF signaling pathway. Similarly, the FC extract exhibited anti-inflammatory effects and modulated TA production, promoting UA exporters (Abcg2/Slc2a9) and enhancing intestinal UA excretion by suppressing the IL-17 signaling pathway. The combination of WC and FC synergistically reduces UA levels by both inhibiting XOD and promoting UA excretion. These results support the potential of WC and FC as natural therapeutic agents for HU management.