OBJECTIVE To clarify the regulatory effect and underlying molecular mechanism of Yishen Qingre Huashi formula(YSQRHS) on chronic nephritis.

METHODS A progressive strategy combining clinical data mining, animal experiment verification and molecular mechanism analysis was adopted. The GSE35487 dataset from the GEO database(5 healthy individuals and 25 nephritis patients) was used to identify differential expressed genes(DEGs) and conduct pathway enrichment analysis. The intersection target genes between nephritis and YSQRHS were retrieved from the HERB database. An adenine-induced chronic nephritis rat model was established, consisting of the normal group, model group, positive control group(Huangkui capsule), and low/high-dose of YSQRHS groups. After the treatments, physiological, urinary and pathological indicators were detected. ELISA was used to detect serum inflammatory factors, while RT-qPCR was employed to verify the expression levels of core genes. Additionally, molecular docking was performed using MOE software to screen potential active components of YSQRHS.

RESULTS Bioinformatics analysis revealed that inflammatory pathways such as NF-κB and TNF were the core pathways involved in nephritis, with FOS, JUN, and GDF-15 identified as key regulation and target genes of YSQRHS. Animal experiments confirmed that YSQRHS could dose-dependently reduce kidney weight and kidney-to-body weight ratio, decrease urinary protein excretion, increase urinary creatinine level, and alleviate glomerular inflammatory cell infiltration, renal tubular necrosis and fibrosis. Meanwhile, it significantly downregulated the serum levels of TNF-α, IL-1β, IL-6 as well as the renal levels of FOS, JUN, GDF-15. Molecular docking results demonstrated that, β-carotene, capsaicin, and other active components could be stably bind to FOS and JUN.

CONCLUSION YSQRHS exerts synergistic anti-inflammatory, anti-fibrotic and renoprotective effects by targeting the NF-κB pathway and its downstream key genes(FOS, JUN and GDF-15). The result of this study provides experimental evidence for its clinical application in the treatment of chronic nephritis.