Objective To observe the effect of Yiqi Tongluo Formula on the protein expression of Wnt4, β-catenin, and Transforming Growth Factor-β1（TGF-β1） in rats’ kidney with unilateral ureteral obstruction（UUO）.Methods Ninety six healthy adult SD rats were randomly divided into sham operation group, model group, low, medium and high dose Yiqi Tongluo Formula groups and losartan group respectively, with 16 rats in each group.Except the sham operation group, unilateral ureteral obstruction renal fibrosis model was established by ligating the left ureter.The sham operation group and model group were given normal saline by gavage, and the low, medium and high dose groups of traditional Chinese medicine and losartan group were given the medicines by gavage according to the corresponding drug concentration; 8 rats in each group were killed at random on the 7 th and 14 th day after modeling, and the blood samples were collected to detect the levels of serum creatinine（SCr） and blood urea nitrogen（BUN）.The pathological changes of renal tissue were observed by HE staining and Masson staining, and the expression of Wnt4, β-catenin and TGF-β1 protein in renal tissues were detected by Western blot.Results The kidney tissue of the sham operation group was normal.Compared with the sham operation group at the same time point, renal interstitial fibrosis was found in the model group, SCr and BUN levels were increased, Wnt4, β-Catenin and TGF-β1 protein expression in renal tissues were significantly increased and the differences were statistically significant（P<0.05）, and with the longer ureteral obstruction, the renal interstitial fibrosis got worse, the renal function reduced more, Wnt4, β-Catenin and TGF-β1 protein expression in renal tissues significantly increased.Compared with the model group at the same time point, the degree of renal interstitial fibrosis was alleviated, Wnt4, β-Catenin and TGF-β1 protein expression in renal tissues significantly decreased, and the differences were statistically significant（P<0.05） in low, medium an high-dose Yiqi Tongluo Decoction group and the Losartan group.Conclusion Wnt/β-catenin signaling pathway is activated in the renal tissues of UUO rats, which may mediate the transcription of TGF-β1.Yiqi Tongluo Formula may inhibit the activation of Wnt/β-catenin signaling pathway and down-regulate the expression of TGF-β1, thus playing an anti-renal fibrosis role.