Tehran University of Medical Sciences Iranian Journal of Allergy, Asthma and Immunology 1735-1502 0 0 2026 09 03 1 20 Zhijian Huang Department of Geriatrics, Quanzhou First Hospital Affiliated to Fujian Medical University, Quanzhou, Fujian, China Youxiao Zeng Department of Geriatrics, Quanzhou First Hospital Affiliated to Fujian Medical University, Quanzhou, Fujian, China Xue’e Huang Department of Geriatrics, Quanzhou First Hospital Affiliated to Fujian Medical University, Quanzhou, Fujian, China Lihong Huang Department of Geriatrics, Quanzhou First Hospital Affiliated to Fujian Medical University, Quanzhou, Fujian, China Yafen He Department of Geriatrics, Quanzhou First Hospital Affiliated to Fujian Medical University, Quanzhou, Fujian, China Yijun Zhou Department of Geriatrics, Quanzhou First Hospital Affiliated to Fujian Medical University, Quanzhou, Fujian, China 2025 11 10 2026 05 09 Heart failure (HF) represents the final stage of multiple cardiovascular disorders. Although LIMK1 regulates cytoskeletal dynamics and contributes to cardiac fibrosis, its exact role in HF remains uncertain. To establish an in vitro HF model, H9c2 cells were treated with isoproterenol (ISO), and LIMK1 expression was analyzed. LIMK1 was silenced using small interfering RNA to investigate its role in cell injury, hypertrophy, fibrosis, cytoskeletal dynamics, and apoptosis. Oxidative stress, reactive oxygen species (ROS), 8-hydroxy-2’-deoxyguanosine (8-OHdG), MDA, SOD, GSH-Px, NADPH oxidase, inflammation, and mitochondrial function were also assessed. Furthermore, the interplay between LIMK1, ROS, and nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB)/NOD-like receptor family pyrin domain containing 3 (NLRP3) signaling was explored using hydrogen peroxide (H2O2; a ROS inducer), diprovocim (an NF-κB activator), and N-acetylcysteine (NAC; a ROS inhibitor). ISO treatment significantly upregulated the expression of LIMK1 in H9c2 cells. Silencing LIMK1 alleviated ISO-induced myocardial cell damage, hypertrophy, and fibrosis, inhibited Cofilin phosphorylation and F-actin expression, and reduced cell apoptosis and ROS production, thereby mitigating oxidative damage. Furthermore, LIMK1 silencing reduced mitochondrial ROS levels, restored mitochondrial membrane potential, and inhibited the release of inflammatory cytokines. However, H2O2 partially reversed the protective effect of LIMK1 silencing. LIMK1 silencing alleviated inflammatory injury by inhibiting NF-κB phosphorylation and NLRP3 expression, and these effects were partially reversed by an NF-κB activator. NAC alleviated cell damage by inhibiting ROS, and its protective effect was also partially reversed by the NF-κB activator. In conclusion, silencing LIMK1 can suppress ROS-mediated activation of the NF-κB/NLRP3 pathway, thereby alleviating cardiomyocyte injury and the inflammatory response. https://ijaai.tums.ac.ir/index.php/ijaai/article/view/4652 https://ijaai.tums.ac.ir/index.php/ijaai/article/download/4652/2415