AIM: To explore the mechanism by which Qingshen granules (QSG) intervene in the microRNA4516 (miR-4516) targeted regulation of the SIAH3/PINK1 axis, enhancing mitophagy and inhibiting renal fibrosis. METHODS: Male SD rats were randomly divided into three groups: control, model, and QSG groups. The QSG aqueous solution was administered via gavage once daily, 4 mL each time, for 8 consecutive weeks. Blood creatinine levels were measured in each group. Hematoxylin-eosin (HE) and Masson staining were utilized to assess the degree of renal pathological damage. Western blot analysis was performed to determine the expression levels of β-actin, PINK1, Parkin, SIAH3, VDAC1, Mfn1, Mfn2, OPA1, LC3B, and P62 proteins in renal tissue. RT-qPCR was used to detect the mRNA expression level of SIAH3 in rat kidney tissue, and transmission electron microscopy was employed to observe mitochondrial damage in renal tissue. QSG-containing serum and transforming growth factor-β1 (TGF-β1) were used to induce an HK-2 cell fibrosis model. The cells were divided into the following groups: normal cell (NC) group, model cell (MC) group, MC+ miR-4516 mimics group, MC+miR-4516 NC+QSG group, MC+miR-4516 mimics+QSG group, and MC+QSG group. Cell activity in each group was detected using the CCK-8 method, and Western blot analysis was performed to determine E-cadherin and α-SMA protein expression levels. The regulation of SIAH3 by miR-4516 was verified using a dual luciferase reporter assay. RT-qPCR was used to detect the mRNA expression of miR-4516, SIAH3 mRNA, and PINK1. RESULTS: The results indicated that QSG intervention reduced fibrosis in rat renal tissue and HK-2 cells, decreased SIAH3 mRNA expression, increased PINK1 expression, and activated mitophagy in renal tissue. In vitro results confirmed that QSG can elevate miR-4516 expression, inhibit SIAH3 mRNA expression, promote PINK1 expression in HK-2 cells, and reduce the expression of the fibrosis marker protein α-SMA. CONCLUSION: In summary, this study preliminarily clarified the mechanism by which QSG intervention targets miR-4516 to regulate the SIAH3/PINK1 axis, thereby enhancing mitophagy and inhibiting renal fibrosis. [ABSTRACT FROM AUTHOR]