Objective To explore the preventive and therapeutic mechanism of Jingfang granules (JF) on hyperuricemia nephropathy induced by potassium oxyzinate (PO) in mice. Methods “Target fishing” strategy was used to prepare magnetic microspheres bound to the effective ingredients of JF. The magnetic microspheres with effective ingredients of JF incubated with normal renal tissue lysates of mice, and the potential binding target proteins were directly captured from the tissue lysates. High-resolution mass spectrometry was used to screen the target proteins that specifically bound to the effective ingredients of JF. The Kyoto Encyclopedia of Genes and Genomes (KEGG) was used to analyze the signaling pathways enriched in target proteins. Thirty-six male ICR mice were randomly divided into the low-dose JF group, mediumdose JF group, high-dose JF group, model group, allopurinol (AL) group, and control group according to body weight, with 6 mice in each group. In addition to intraperitoneal injection of 250 mg/kg PO solution, mice in the low-, medium-, and high-dose JF groups were gavaged with 0. 5,1, and 2 g/kg JF, respectively; mice in the model group were gavaged with corresponding volume of purified water, mice in the AL group were gavaged with 25 mg/(kg·d) AL solution, and mice in the control group were gavaged with corresponding volume of purified water. All were administered once a day for 4 consecutive weeks. One hour after the last administration, blood was taken to measure serum uric acid (UA), urea nitrogen (BUN), and creatinine (Cr). Renal tissue was observed by H&E staining. Main indicators of oxidative stress, such as malonaldehyde (MDA), superoxide dismutase (SOD), and glutathione peroxidase (GSH) were detected by ELISA. Main indicators of target protein-related signaling pathways (oxidative stress proteins including Nrf2, HO-1 and NQO1, and mucosal barrier functional proteins including Occludin and ZO-1) were detected by Western blotting. Results Totally 176 specific binding proteins were identified from the renal tissues, and the function of target proteins could be related to peroxisome, glutathione metabolism, tight junction, adhesive junction, and other pathways. The serum levels of UA, BUN, and Cr in the model group were significantly higher than those in the control group (all P<0. 05). The levels of serum UA, BUN, and Cr in the JF and AL groups were significantly lower than those in the model group, and the JF group showed a significant dose dependence (P<0. 05) . The renal cortex of mice in the medium- and high-dose JF groups tended to be normal, the shape of renal tubules improved, and inflammation was significantly alleviated, especially in the high-dose JF group. There was a small amount of inflammatory cell infiltration in the interstitium of mice in the AL group, which was improved compared with that of the model group. Compared with the control group, MDA content increased, SOD and GSH activities decreased (all P<0. 05), the relative expression levels of Nrf2, HO-1 and NQO1 proteins decreased, and the relative expression levels of Occludin and ZO-1 proteins decreased in the tissues of mice in the model group. Compared with the model group, MDA content decreased, SOD and GSH activities increased (all P< 0. 05), the relative expression levels of Nrf2, HO-1 and NQO1 proteins increased, and the relative expression levels of Occludin and ZO-1 protein increased in JF groups and AL group (all P<0. 05). Significant differences were found in various indicators between the high-dose JF and low-dose JF groups (all P<0. 05). Conclusions The target protein of effective ingredients of JF in kidney tissues may be related to peroxisome, glutathione metabolism, tight junction and adhesive junction. JF have preventive effect on hyperuricemia nephropathy in mice in a dose-dependent manner, and their effects may be related to inhibiting oxidative stress and improving mucosal barrier. [ABSTRACT FROM AUTHOR]