Objective To explore the effect of Jianpi Shenshi Formula preotecting kidney damage and regulating intestinal flora in uricase-deficient(Uox-/-) hyperuricemia rats,and to provides a scientific basis for the clinical application for Jianpi Shenshi Formula. Methods An animal model of uricase-deficient (Uox-/-) hyperuricemia was established by using of CRISPR/Cas9 technique,dividing into uricase-deficient hyperuricemia group (Uox-/-) and JPSSF group (12.50 g·kg-1·d-1), wild-type SD rats(WT) were used as the control group. All animals were treated orally for 60 days. Renal function indexes (UA, UREA, CREA) from serum were detected by an automatic biochemical analyzer; Coomassie brilliant blue method was used to assess urinary protein for 24 h; HE staining was applied to assess kidney histopathological damage; Level of Uric acid from intestinal tissue was determined by phosphotungstic acid method; The qPCR was used to detect the expression levels of urate transporter gene; The microecological diversity of intestinal flora was determined by 16S rRNA V4 region sequencing. Results Here, we showed that Jianpi Shenshi Formula significantly decreased renal function indexes (UA,UREA, CREA) and alleviated renal lesions damage. However, no significant influence on uric acid from the colonic tissue (P>0.05). Down-regulated urate transporter URAT1 mRNA expression in JPSSF group. The PCoA and NMDS analysis have showed that the hyperuricemia causes disorders to the intestinal flora, while the Jianpi Shenshi Formula showed little effect on reconstruction of the intestinal flora(P>0.05). At the genus level, compared with the Uox-/- group, there were certain increase in Lactobacillus， Proteus， Prevotellaceae NK3B31 group in JPSSF group, Clostridium sensu stricto 1 and Bifidobacterium were decreased in JPSSF group.However,there were no significant differences between Uox-/- group and JPSSF group (P>0.05). Conclusion Our results indicated that Jianpi Shenshi Formula notably ameliorated hyperuricemia and related renal damage caused by the deposition of uric acid crystals, while has no obvious effect on the intestinal uric acid metabolism and intestinal microecological disorders.