Objective Clinical evidence suggests that the risk of hyperuricemia is increased under high temperature-humidity conditions. The current study aimed to explore this effect and the possible molecular mechanism responsible for the environmental effect on serum uric acid (SUA) levels, using a recently established hyperuricemia rat model. Methods Hyperuricemia was induced in rats by potassium oxonate (PO group) or a high-purine diet (HPD group) using traditional chemical-induction method. The rats were also irradiated in a high temperature-humidity incubator (37℃, 80% humidity) for 1 h/ d to produce the high temperature-humidity group (HTH group) and high purine diet +high temperature-humidity group (HPD + HTH group). A control group (CON group) was also included. The experiment lasted for 12 weeks. Anal temperature and body weight were measured during daily irradiation, and blood samples were collected every 4 weeks to examine SUA and creatine levels in each group. After the experiment, the kidneys were removed and expression levels of ATP-binding cassette super family G2 ( ABCG2) and urate transporter 1 ( URAT1) were determined by Western Blot and immunohistochemistry. Results The anal temperature and dehydration rate of rats in the HTH and HPD + HTH groups were significantly increased after irradiation in a high temperature-humidity incubator (P<0. 05). SUA levels were significantly higher in the HTH group compared with the CON and PO groups, and in the HPD +HTH compared with the HPD group (P< 0. 05). There was no significant difference in serum creatinine among the five groups after 12 weeks (P> 0. 05). Expression levels of the uric acid secretion protein ABCG2 were significantly lower in the HTH and HPD + HTH groups compared with the CON group, in the HTH group compared with the PO group, and in the HPD + HTH group compared with the HPD group, as shown by Western Blot (P< 0. 05). Expression levels of the urate transporter URAT1 were significantly higher in the HTH and HPD + HTH groups compared with the CON group, in the HTH group compared with the PO group, and the HPD + HTH group compared with the HPD group (P< 0. 05). Immunohistochemistry indicated that both ABCG2 and URAT1 were expressed in proximal convoluted tubule epithelial cells. ABCG2 expression levels were lower while URAT1 expression levels were higher in the HPD + HTH and HTH groups compared with the other groups (P< 0. 05). Conclusions A high temperature-humidity environment can influence the secretion and reabsorption of uric acid by disrupting the function of the uric acid transporters ABCG2 and URAT1, which are located in the proximal tubules of the kidney. This can lead to increased SUA levels, potentially aggravating the occurrence and development of hyperuricemia.