Objective To evaluate the protective effects of the traditional Chinese medicine formula Shenxiankang on renal injury and fibrosis, and to explore its potential mechanisms of action. Methods Chronic kidney disease (CKD) model was established in mice using unilateral ureteral obstruction (UUO). The mice were randomly divided into four groups: sham, UUO, and Shenxiankang(SXK) Low/ High dose groups (1500, 4500 mg /(kg·d)), each comprising eight mice. The each SXK groups received daily oral administration of Shenxiankang, and the remaining mice were gavaged equivalent volumes of saline for 7 d. After the experiment, renal tissues were collected for assessment of renal injury and fibrosis using HE and Masson staining. The expression levels of fibrosis markers and proteins involved in the epithelial membrane protein 3 (Emp3) and Tgf-β/ Smad3 signaling pathway were determined by Real-time PCR, immunohistochemistry, and Western Blot. In cell-based experiments, the effects of Shenxiankang on the Emp3 / Tgf-β/ Smad3 pathway and its interaction with TGF-beta receptor R2 (Tgfβ2) were further analyzed using an Emp3 knockdown and Co-IP assays. Results Shenxiankang significantly reduced immune cell infiltration and tubular atrophy in the UUO model group and decreased the expression of kidney injury markers kidney injury molecule 1 (Kim1) and Lipocalin 2 (Lcn2), confirming its efficacy in alleviating renal injury. Masson staining and analysis of fibrosis markers Fibronectin (Fn) and α-smooth muscle actin (α-SMA) indicated that Shenxiankang effectively suppressed fibrosis induced by UUO. Mechanistic studies revealed that Shenxiankang exerted its effects by selectively downregulating the abnormal activation of the Emp3 / Tgf-β/ Smad3 signaling pathway, a finding further supported by cellular experiments showing that Shenxiankang modulates Tgf-β/ Smad3 signaling through Emp3 regulation. Moreover, the Co-IP experiment result indicate that Shenxiankang exerts its effects by regulating the interaction between Emp3 and Tgfβ2. Conclusions Shenxiankang exhibits significant protective effects in a mouse model of chronic kidney disease, effectively reducing renal injury and fibrosis. These effects are likely mediated through the downregulation of the Emp3 / Tgf-β/ Smad3 signaling pathway, suggesting Shenxiankang ’ s potential therapeutic value in renal protection.