Objective To explore the molecular mechanism of Wenshenxingpihuazhuo formula on delaying progression of chronic renal failure (CRF), and provide theoretical base in treating chronic renal failure. Methods An rat model of CRF were induced using a single intravenous vein injection with adriamycin (6 mg/kg), the rat treated with adriamycin were divided into five groups: normal group, model group, lowdose group, middle dose group and high dose group. After administration of Wenshenxingpihuazhuo formula, the coomassie brilliant blue method was used to assess 24 h urinary protein; level of uric acid, urea, and creatinine in serum were detected using an automatic biochemical analyzer; kidney histopathological changes were measured by H＆E staining. The mRNA expression in renal fibrosis-related signaling (TIMP-1, MMP-9, TGF-β1, Col4a2R and Fn1) was examined by q-PCR analysis. Results Oral administration of Wenshenxingpihuazhuo formula markedly decrease the 24 h urinary protein and renal index, significantly reduced the level of uric acid, urea, and creatinine from serum, obviously alleviated the renal pathological damage, including infiltration of inflammatory cell into kidney tissue, hyperplasia of fibrous tissue and glomerular deformation and atrophy. Furthermore, the mRNA expression of TIMP-1, TGF-β1, Col4a2R, Fn1 in Wenshenxingpihuazhuo formula treated group were decreased as compared to model group (P<0.05 or P<0.01), while MMP-9 transcriptional expression was upregulated (P<0.05). Conclusion Wenshenxingpihuazhuo formula could delay renal fibro and progression of chronic renal failure via regulating transcriptional expression in kidney fibrosis-related signaling, including recover the balance MMP9 and TIMP-1 and downregulation of TGF-β1, Col4a2R and Fn1.