Inhibition of Calpain-1 ameliorates doxorubicin-induced cardiotoxicity by suppression of p53-mediated ferroptosis.

Authors

Zhuo Zhang, Xinbin Zheng, Dabao Xiao, Mengya Zeng, Junru He, Shaowu Xiao, Maozhong Yao, Yuewu Chen

Published in

Toxicology and applied pharmacology. Pages 118001. Aug 17, 2026. Epub Aug 17, 2026.

Abstract

Doxorubicin (Dox), a widely applied anthracycline antitumor agent, exhibits severe dose-dependent cardiotoxicity that greatly restricts its clinical utilization, yet the molecular mechanism linking Calpain-1 to cardiomyocyte ferroptosis in doxorubicin-induced cardiotoxicity (DIC) remains poorly clarified. This study aimed to explore how Calpain-1 regulates cardiomyocyte ferroptosis in DIC. Differentially expressed proteins from Dox-treated H9c2 cells were screened by proteomics, stable Calpain-1 knockdown cell lines were generated to detect cell viability and ferroptosis indicators through CCK-8, FerroOrange staining, biochemical kits and western blotting, and PPI network, co-IP and cycloheximide chase assays were performed to confirm Calpain-1-p53 binding and p53 protein stability regulation, with p53 rescue tests further verifying their functional relationship. In vivo, Calpain-1 expression was detected in Dox-exposed mouse hearts, and AAV9-driven myocardial Calpain-1 knockdown was established in DIC mice, where myocardial injury and ferroptosis were evaluated by HE staining, serum CK-MB/LDH and protein detection. Calpain-1 was significantly upregulated in Dox-stimulated cardiomyocytes and cardiac tissue. Calpain-1 silencing alleviated Dox-induced ferroptosis in vitro, with decreased Fe²⁺, MDA, LPO and ACSL4, elevated GSH and SLC7A11; co-IP verified direct Calpain-1-p53 interaction, and cycloheximide assays proved Calpain-1 inhibits p53 degradation to accumulate p53 protein, whereas p53 overexpression completely abolished the protective effect of Calpain-1 knockdown. Consistent with cellular results, cardiac-specific Calpain-1 knockdown mitigated myocardial damage and suppressed ferroptosis in DIC mice. These findings suggest that Calpain-1 exacerbates DIC by stabilizing p53 to initiate p53-dependent cardiomyocyte ferroptosis, offering a novel therapeutic target and theoretical basis for DIC intervention.

PMID:
42607844
Bibliographic data and abstract were imported from PubMed on 18 Aug 2026.

Read full publication at:
Please sign in to see all details.

Stats

1-terrible, 9-excellent. How would you rate this publication? Sign in in to submit your rating.

Post a comment

You need to be signed in to post comments. You can sign in here.

Comments

There are no comments yet.