Astilbin is a flavonoid compound derived from the rhizome of Smilax china L. The effects and possible molecular mechanisms of astilbin on potassium oxonate-induced hyperuricemia mice were investigated in this study. Different dosages of astilbin (5, 10, and 20 mg/kg) were administered to induce hyperuricemic mice. The results demonstrated that the serum uric acid (Sur) level was significantly decreased by increasing the urinary uric acid (Uur) level and fractional excretion of urate (FEUA) with astilbin, related with suppressing role in meditation of Glucose transporter 9 (GLUT9), Human urate transporter 1 (URAT1) expression and up-regulation of ABCG2, Organic anion transporter 1/3 (OAT1/3) and Organic cation transporter 1 (OCT1). In addition, kidney function parameters, including serum creatinine (Scr) and blood urea nitrogen (BUN) were restored in astilbin-treated hyperuricemic rats. Further investigation indicated that astilbin prevented the renal damage against the expression of Thioredoxin-interacting protein (TXNIP) and its related inflammation signal pathway, including NLR pyrin domain-containing 3/Nuclear factor κB (NLRP3/NF-κB), which is associated with the up-regulation of interleukin-1β (IL-1β) and interleukin-18 (IL-18), and also presented a renal protective role by suppression oxidative stress. Moreover, astilbin inhibited activation of the Janus kinase 2/signal transducer and activator of transcription 3 (JAK2/STAT3) cascade and over-expression of suppressor of cytokine signaling 3 (SOCS3) in the kidneys of potassium oxonate-induced mice. These findings provide potent evidence and therapeutic strategy for astilbin as a safe and promising compound in the development of a disease-modifying drug due to its function against hyperuricaemia and renal injury induced by potassium oxonate.