OTUD1 Relieves Coxsackievirus-Induced Ferroptosis and Inflammation in Myocardial Cells by Stabilizing NRF2 and Inhibiting NF-κB Signaling

Viral myocarditis (VMC) is a common inflammatory myocardial disease in children and is associated with heart failure and sudden cardiac death. Coxsackievirus B3 (CVB3) is the most common pathogen, and inflammation and ferroptosis are the key pathological events of CVB3-induced myocarditis. Studies indicate that the deubiquitinase OTUD1 is involved in cardiac inflammation and ferroptosis, but its role in CVB3-induced myocardial inflammation and ferroptosis remains unclear. Therefore, this study aimed to investigate the function and mechanism of OTUD1 in inflammation and ferroptosis in myocardial cells in the context of CVB3 infection. Mice and H9c2 cells infected with the CVB3 strain were used to construct in vivo and in vitro CVB3 myocarditis models. The inflammatory cytokines IL-1β, IL-6, and TNF-α; Fe2+; MDA; SOD; and the myocardial injury markers CK-MB and cTnI were detected using assay kits. Cell viability was measured by a CCK-8 assay, myocardial injury was evaluated by HE staining, and the expression of key proteins was detected by western blotting, immunohistochemistry, and immunofluorescence. After CVB3 infection, heart tissue was damaged; the levels of CK-MB and cTnI increased; the viability of H9c2 cells decreased; the levels of IL-1β, IL-6, TNF-α, Fe2+, MDA, and ROS increased; the level of SOD decreased; and the expression of the ferroptosis-related proteins GPX4 and SLC7A11 decreased. Second, CVB3 infection also repressed OTUD1 expression in mouse heart tissues and H9c2 cells. Importantly, the above effects of CVB3 were partially attenuated by overexpression of OTUD1, indicating that OTUD1 can inhibit CVB3-induced myocardial ferroptosis and inflammation. In addition, CVB3 infection also repressed the expression of NRF2 in vivo and in vitro and promoted the activation of NF-κB signaling. A mechanistic study revealed that OTUD1 could relieve CVB3-induced inflammation and ferroptosis by stabilizing NRF2 expression and inhibiting NF-κB signaling activation. Our study revealed the potential of OTUD1 to ameliorate inflammation and ferroptosis in myocardial cells induced by CVB3 infection, providing a potential intervention target for the treatment of VMC.