ABSTRACTSmilax glabra Roxb. is a natural herb, exhibits significant uric acid lowering effect in clinical. However, the specific mechanism remains to be clarified. To study this problem, intraperitoneal injection of Potassium oxazinate (PO) and Hypoxanthine (HX) induced hyperuricemia in mice model, and mice were divided into control, model, and model plus ethanol extract (TFL), ethyl acetate extract (TFL_Y), n‐butanol extract (TFL_Z), and residual aqueous extract (TFL_S) of Smilax glabra Roxb., And then a follow‐up test. Furthermore, possible pharmacological components were detected by UPLC‐Q‐TOF‐MS. The results showed that serum creatinine, blood urea nitrogen, d‐lactate, lipopolysaccharide levels and xanthine oxidase. They could upregulate renal OAT1 and ABCG2, downregulate renal URAT1, reduce renal tubular dilation and interstitial inflammatory cell infiltration. Moreover, they could upregulate intestinal tight junction proteins, increase the number of intestinal goblet cells, and repair damage to the intestinal barrier. Moreover, 16S rRNA sequencing analysis showed that restored intestinal microbiota by increasing probiotic bacteria (Candidatus Saccharimonas, Lachnospiraceae NK4A136_group) and reducing pathogenic bacteria (Helicobacter). Furthermore, it was found that all control taint flavonoids such as Neoastalbin, Astibin, Neoiosastilbin, Isoastalbin, Engeletin, and Isoengeletin.