Highlights
- •This study reported the role of Fyn in diabetic nephropathy (DN).
- •We proposed Fyn as a potential therapeutic target against DN.
- •Fyn deficiency exerted antioxidant effects by activating the Sirt1/Foxo3a pathway.
- •Fyn facilitated the bond of c-Cbl and Sirt1 by phosphorylation at Tyr731 of c-Cbl.
- •c-Cbl promoted K48-linked polyubiquitination at Lys377 and Lys513 of Sirt1.
Abstract
Objective
Oxidative stress (OS) is the main cause leading to diabetic renal fibrosis. Recently,
Fyn was paid much attention on OS and emerged as a pivotal player in acute kidney
injury, while whether Fyn regulates oxidative stress in chronic diabetes nephropathy
(DN) has not been clarified yet. The purpose of this study was to identify the role
of Fyn in DN and elucidated its regulatory mechanism.
Methods
The db/db mice and littermate control C57BKS/J mice were injected by tail vein with Fyn interfering
adenovirus or Fyn overexpressing adenovirus to investigate the role of Fyn in vivo. Primary glomerular mesangial cells (GMCs) were used for in vitro studies.
Results
Fyn was up-regulated in high glucose (HG)-induced GMCs and kidneys of diabetic mice.
Additionally, Fyn knockdown reduced the level of OS in HG-induced GMCs and kidneys
of diabetic mice, thereby ameliorating diabetic renal fibrosis. While overexpression
of Fyn significantly increased the level of OS in GMCs and kidney tissues, resulting
in renal damage. Moreover, Fyn deficiency exerted antioxidant effects by activating
the Sirt1/Foxo3a pathway. Mechanistically, Fyn facilitated the combination of c-Cbl
and Sirt1 by phosphorylating c-Cbl at Tyr731, which triggered K48-linked polyubiquitination of Sirt1 at Lys377 and Lys513 by c-Cbl and promoted Sirt1 degradation, impairing the antioxidant effects of Foxo3a.
Conclusions
Fyn deficiency promoted Foxo3a nuclear transcription via reducing the ubiquitination of Sirt1 by c-Cbl, thereby alleviating renal oxidative
damage in diabetic mice. These results identified Fyn as a potential therapeutic target
against DN.
Abbreviations:
24 h UP (24-h urine protein), BUN (blood urea nitrogen), CHX (Cycloheximide), Cr (serum creatinine), DHE (Dihydroethidium), DN (diabetic nephropathy), ECM (extracellular matrix), FBG (fasting blood glucose), FN (fibronectin), GMCs (glomerular mesangial cells), GSP (glycated serum protein), HE (hematoxylin-eosin), HEK (Human embryonic kidney), HG (high glucose), ICAM-1 (intercellular cell adhesion molecular-1), IF (immunofluorescence), IHC (immunohistochemistry), IP (immunoprecipitation), KW/BW (renal weight/body weight ratio), LPO (lipid peroxide), MDA (malondialdehyde), NAC (N-Acetyl-L-cysteine), NC (negative control), NG (normal glucose), OS (oxidative stress), PAS (Periodic Acid-Schiff), ROS (reactive oxygen species), SD (Sprague-Dawley), Si-RNA (small interfering RNA), SOD (superoxide dismutase), SPF (specific pathogen-free)Keywords
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Article info
Publication history
Published online: December 17, 2022
Accepted:
December 8,
2022
Received:
August 30,
2022
Identification
Copyright
© 2022 Published by Elsevier Inc.
