Abstract： Objective： To investigate the effect and possible mechanism of heart three needles[needling at the Baihui （GV20）， Danzhong （CV17）， and bilateral Shenmen（HT7）points] in alleviating myocardial injury following myocardial ischemia-reperfusion injury （MIRI）. Methods： Thirty healthy male mice were randomly divided into a sham operation group，a model group，and a heart three needles（ HTN） group，with 10 mice in each group. Except for the sham operation group，MIRI models were established in the other two groups by ligating the left anterior descending coronary artery followed by 30 minutes of reperfusion. After successful modeling， the HTN group received electroacupuncture （EA） at Baihui （GV20）， Danzhong （CV17）， and bilateral Shenmen （HT7） points with dense-sparse waves （frequency 2 Hz/100 Hz） and current intensity of 2 mA，30 minutes per session，once daily for 30 consecutive days. The sham operation group and the model group received only routine feeding without EA intervention. After the intervention， cardiac function was assessed by echocardiography； serum levels of creatine kinase-MB（ CK-MB） and cardiac troponin I（ cTnI） were detected by ELISA；myocardial histopathological morphology was observed by HE staining；myocardial collagen deposition and fiber arrangement were analyzed by Masson staining； iron deposition was observed by Prussian blue （Perls） staining； protein expressions of ferritin heavy chain （FTH）， ferritin light chain （FTL），and glutathione peroxidase 4 （GPX4） in myocardial tissue were detected by Western blot； mRNA expressions of ferroptosis pathway-related genes were detected by RT-qPCR. Results：Compared with the sham operation group， the model group showed decreased left ventricular ejection fraction （LVEF） and left ventricular fractional shortening （LVFS） （P＜0.05）， elevated serum CK-MB and cTnI levels （P＜0.05）， inflammatory infiltration，structural disorganization，and collagen deposition in myocardial tissue，increased iron ion aggregation， and downregulated protein and mRNA expressions of GPX4，FTH，and FTL（ P＜0.05）. After intervention，compared with the model group，the HTN group showed increased LVEF and LVFS （P＜0.05），decreased serum CK-MB and cTnI levels （P＜0.05）， significantly improved myocardial pathological injury， reduced iron deposition， and upregulated protein and mRNA expressions of GPX4，FTH，and FTL （P＜0.05）. Conclusion：Heart three needles can improve cardiac function and myocardial injury in MIRI mice and inhibit ventricular remodeling. The mechanism may be related to regulating the ferroptosis pathway and upregulating the expressions of GPX4，FTH，and FTL.