Proteomic Characterization of Aprotinin’s Immune-Modulating Role in Renal Fibrosis Following UUO

 

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https://storage.unitedwebnetwork.com/files/1099/7bdef844463231b1f6a04fcefe268685.pdf
Proteomic Characterization of Aprotinin’s Immune-Modulating Role in Renal Fibrosis Following UUO

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Qinyuan
Deng
Qinyuan Deng deng835552032@gmail.com The First People’s Hospital of Yunnan Province Department of Nephrology Kunming China * The Affiliated Hospital of Kunming University of Science and Technology Department of Nephrology Kunming China
Ying Shen 1402202854@qq.com The First People’s Hospital of Yunnan Province Department of Nephrology Kunming China - The Affiliated Hospital of Kunming University of Science and Technology Department of Nephrology Kunming China
Lu Song songlu1995@163.com The First People’s Hospital of Yunnan Province Department of integrated traditional Chinese and Western Medicine Kunming China - The Affiliated Hospital of Kunming University of Science and Technology Department of integrated traditional Chinese and Western Medicine Kunming China
Yutaka Kakizoe kakizoe@kumamoto-u.ac.jp Kumamoto University Graduate School of Medical Sciences Department of Nephrology Kunming China -
Fucheng Luo luofc@lpbr.cn Institute of Primate Translational Medicine, Kunming University of Science and Technology State Key Laboratory of Primate Biomedical Research Kunming China -
Fei Chen Cf13008690576@163.com The First People’s Hospital of Yunnan Province Department of integrated traditional Chinese and Western Medicine Kunming China - The Affiliated Hospital of Kunming University of Science and Technology Department of Nephrology Kunming China
Jian Xu xujian3979@sina.com The First People’s Hospital of Yunnan Province Department of integrated traditional Chinese and Western Medicine Kunming China - The Affiliated Hospital of Kunming University of Science and Technology Department of Nephrology Kunming China
Xinyu Wang wangxinyu_mat@126.com The First People’s Hospital of Yunnan Province Department of Nephrology Kunming China - The Affiliated Hospital of Kunming University of Science and Technology Department of Nephrology Kunming China
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Chronic kidney disease (CKD) is characterized by progressive renal fibrosis and immune dysregulation. Aprotinin, a broad-spectrum serine protease inhibitor, has shown renoprotective effects in acute kidney injury but its role in chronic obstructive nephropathy remains unclear. We investigated aprotinin’s antifibrotic and immunomodulatory effects in a murine model of unilateral ureteral obstruction (UUO).

Male C57BL/6 mice were subjected to UUO and treated with aprotinin (0.5 or 1 mg/day) via subcutaneous osmotic pumps for 7 days. Kidney injury and fibrosis were assessed histologically and by measuring serum creatinine, BUN, and expression of fibrosis- and inflammation-related genes. Proteomic profiling (LC-MS/MS) and immune deconvolution (CIBERSORT) were used to elucidate molecular and immunological mechanisms.

High-dose aprotinin (1 mg/day) significantly improved tubular injury and reduced interstitial fibrosis in the obstructed kidney. Despite histological improvement, BUN and creatinine were mildly elevated, possibly due to effects on the contralateral kidney. Proteomic analysis revealed 79 differentially expressed proteins linked to immune regulation. Notably, aprotinin downregulated Cathepsin S (CTSS), a key protease involved in antigen presentation and CD4⁺ T cell differentiation. CIBERSORT analysis showed that aprotinin partially restored naive CD4⁺ T cells and reduced T follicular helper (Tfh) cells, indicating a shift in immune cell composition. Western blot confirmed reduced CTSS expression. Functional enrichment suggested that aprotinin modulates immune responses and mitochondrial metabolism.

Aprotinin attenuates renal fibrosis in UUO via CTSS-mediated immunomodulation, particularly by altering CD4⁺ T cell dynamics. These findings underscore the therapeutic potential of serine protease inhibitors in immune-driven kidney disease and highlight CTSS as a promising molecular target. A schematic summarizes aprotinin’s immunomodulatory effects in UUO, highlighting CTSS downregulation and restoration of CD4⁺ T cell balance.

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