As the newly discovered programmed cell death is characterized by the accumulation of lipid peroxides, iron-dependent ferroptosis is entirely distinct from apoptosis. The anti-apoptotic properties of Kindlin-2 have been reported in hepatocytes, NP cells, tumor cells, fibroblasts, etc., yet its protective role in ferroptosis remains undetermined. Our study first deciphered the underlying mechanism of Kindlin-2 in cardiac ferroptosis. The Kindlin-2 level was decreased in cardiac tissues of I/R mice, ICM patients, and cardiomyocytes underwent hypoxia stimulation. Cell viability of cardiomyocytes is positively regulated by Kindlin-2. In response to tail intravenous injection of AAV9 Fermt2 in mice, cardiomyocyte-specific Kindlin-2 overexpression highly alleviates I/R injury through inhibiting cardiomyocyte ferroptosis in vivo, while cardiomyocyte-specific low expression of Kindlin-2 can impair cardiac functions, which are accompanied by cardiomyocyte ferroptosis. Meanwhile, experiments in vitro verified that Kindlin-2 prevented the ferroptosis in cardiomyocytes treated by H/R and endogenous deficiency of Kindlin-2 in cardiomyocytes has been subsequently identified to induce ferroptosis spontaneously without exogenous stimulation. Mechanistically, Kindlin-2 accelerates the interaction between Otub1 and Slc7a11. Consequently, deubiquitinated Slc7a11 contributes to activate GSH and Gpx4 to exert the anti-ferroptosis effect. Slc7a11/GSH/Gpx4 cascades strengthened by Kindlin-2 can be abolished by the Otub1 KD. Therefore, The prevention effect of Kindlin-2 in cardiac ferroptosis and dysfunctions is Otub1-dependent and Otub1 rescued cardiomyocyte ferroptosis and cardiac injury deteriorated by Kindlin-2 deficiency. Collectively, our research reveals that Kindlin-2/Otub1/Slc7a11 cascade improves cardiac I/R injury through inhibiting ferroptosis and may be the potential therapeutic target of ischemic cardiomyopathy.