Terminalia chebula Retz. extract relieves gout arthritis by inhibiting xanthine oxidase, the uric acid transporter, and NLRP3 inflammasome activation

Abstract

Ethnopharmacological relevance: Gout is a metabolic disorder accompanied by high serum uric acid levels and joint inflammation due to disturbances in purine metabolism in the body. The dried fruit of Terminalia chebula Retz. is recorded in the "Four Medical Tantras" for the treatment of gout and the core anti-gout component of the Tibetan clinical prescription, such as TongFengTangSan. However, the anti-gout efficacy has not been reported yet.

Aim of study: To evaluate the anti-gout effect and mechanisms of Terminalia chebula Retz. in gout model rats.

Materials and methods: First, the components of the Terminalia chebula Retz. extract were detected and characterized using ultra performance liquid chromatography with quadrupole time-of-flight mass spectrometry technology. A gout model was established using the continuous intragastric administration of 200 mg/kg of potassium oxonate and 300 mg/kg of hypoxanthine for 44 days, and 8 mg monosodium urate suspension was injected once in the joint cavity on the 42nd day. One hour after modeling, Terminalia chebula Retz. extract was administered by gavage at low, medium, and high doses. The corresponding biochemical indicators at the protein and gene levels were detected by real-time quantitative polymerase chain reaction (RT-qPCR) and western blot.

Results: A total of 149 compounds, comprising 23 phenolic acids, 104 tannins, 5 flavonoids, 14 terpenoids, and three other compounds, were identified in Terminalia chebula Retz. extract using the ultra performance liquid chromatography with quadrupole time-of-flight mass spectrometry method. The in vivo pharmacodynamics experiments showed that Terminalia chebula Retz. extract significantly reduced the serum uric acid level, the ankle swelling level, and the level of inflammatory factors in the gout rats. Terminalia chebula Retz. extract also decreased the serum xanthine oxidase, alanine aminotransferase, aspartate aminotransferase and diamine oxidase activity of the gout rats. The western blot and PCR experiments showed that treatment with Terminalia chebula Retz. extract down-regulated the mRNA and protein levels of urate transporter 1 and glucose transporter 9 in the kidney tissues. An immunofluorescence experiment revealed that Terminalia chebula Retz. extract strengthened the intestinal barrier by the up-regulation on the protein expression of occludin and zonula occludens-1 in the ileum. In addition, Terminalia chebula Retz. extract was found to alleviate inflammation by inactivating the renal NOD-like receptor family pyrin domain-containing 3 (NLRP3) inflammasome and the synovial membranes of joints. Terminalia chebula Retz. treatment down-regulated the protein or mRNA levels of NLRP3 inflammasome family members, including toll-like receptor 4, toll-like receptor 2, NLRP3, nuclear factor kappa-B, apoptosis-associated speck-like protein containing a CARD and interleukin-1β.

Conclusion: This study demonstrated that Terminalia chebula Retz. extract alleviated gout symptoms through the dual effects of lowering UA and relieving inflammation through inhibiting NLRP3 inflammasome activation.

Keywords: Gout; NLRP3 inflammasome; Terminalia chebula Retz.; Uric acid transporter.