Objective : To investigate the effect of palmitoleic acid (POA) on pyroptosis of cardiomyocytes after hypoxia/reoxygenation⁃induced injury in the human myocardium . Methods : The experiment comprised a control group (Control , normal culture) , a hypoxia/reoxygenation group (HR) , a palmitoleic acid⁃treated group ( HR + POA) , and an anhydrous ethanol control group ( HR + ET) . Cardiomyocytes viability was assessed using CCK⁃8 assay , and the level of pyroptosis of cardiomyocytes was measured through the double staining with Hoechst33342/ PI and LDH assay . ELISA was employed to detect the release of inflammatory factors IL⁃1β and IL⁃18 in the cell culture supernatant . qRT⁃PCR and Western blot were utilized to determine the relative expression levels of mRNA and protein of pyroptosis⁃related genes , namely NLRP3 , ASC , Caspase⁃1 , GSDMD , IL⁃1β and IL⁃18 , respectively . Results : CCK⁃8 assay showed that the survival of hypoxic/reoxygenated cardiomyocytes increased with the addition of POA at concentrations ranging from 25 to 100 μmol/L , as compared to the HR group ; a hypoxia/reoxy⁃genation model of cardiomyocyte was established . The expression of protein and mRNA increased in NLRP3 , ASC ,Cleaved caspase⁃1 , GSDMD⁃N , IL⁃Iβ and IL⁃ 18 vs the control group(P < 0. 05) , the positive percentage of Hoechst33342/PI staining in cardiomyocytes increased significantly (P < 0. 05) , the release of LDH , IL⁃Iβ , and IL⁃18 increased (P < 0. 05) . After intervention with 100 μmol/L POA , the protein and mRNA expression levels of NLRP3 , ASC , Cleaved caspase⁃1 , GSDMD⁃N , IL⁃Iβ , and IL⁃18 were significantly reduced in the HR + POA group vs HR + ET group( P < 0. 05) . The positive percentage of Hoechst33342/PI staining in cardiomyocytes decreased significantly , and the levels of LDH , IL⁃Iβ and IL⁃18 significantly decreased (P < 0. 05) . Conclusion Palmitoleic acid may alleviate hypoxia/reoxygenation⁃induced injury of cardiomyocytes by inhibiting pyroptosis and inflammatory response after hypoxia/reoxygenation in human myocardium .