Objective To prepare nanocarriers capable of targeting the hypoxia-injured cardiomyocytes and to investigate its physicochemical characteristics, cytotoxicity and targeted binding to injured primary cardiomyocytes. Methods The dendrimer nanocarriers (non-targeting nanocarriers) and angiotensin Ⅱ Type 1 (AT1)-modified dendrimer nanocarriers (targeting nanocarriers) were synthesized. The chemical structure, gene compact capacity, particle size and zeta potential of the targeting nanocarriers were determined and the morphology was observed. The toxicity to cardiomyocytes was detected. The primary cardiomyocytes of neonatal Sprague Dawley (SD) rat were randomly divided into 3 groups: hypoxic injury + non-targeted group (The primary cardiomyocytes were cultured with non-targeting nanocarriers under hypoxic conditions), hypoxic injury + targeted group (The primary cardiomyocytes were cultured with targeting nanocarriers under hypoxic conditions), normal + targeted group (The primary cardiomyocytes were cultured with targeting nanocarriers under normoxic conditions). The targeted adhesion of the nanocarriers to injured cardiomyocytes and normal cardiomyocytes were observed (with Laser scanning confocal microscope) and quantified (by flow cytometry). Results Non-targeting nanocarriers and targeting nanocarriers were successfully synthesized. When the weight ratio of targeting nanocarriers to gene was 6: 1, the gene was completely incorporated, with 180±55 nm in size and + 5.4 mV in zeta potential, which posessing homogeneous spherical shape under transmission electron microscopy. When the concentration was less than 200 μg/mL, the cell viability was more than 85% at different time points. Compared with hypoxic injury + non-targeted group (7102±134.3) and normal + targeted group (8120±234.7), the intensity of red fluorescence (targeted adhesion efficiency) of myocardial cell membrane was significantly enhanced in hypoxic injury + targeted group (12350±494.9) (P <0.05). Conclusion AT1-modified dendrimer nanocarrier was successfully prepared, which can effectively binding to the injured cardiomyocytes overexpressing angiotensin Ⅱ type 1 receptor (AT1R) with low cytotoxicity in vitro. It may provide a new targeted delivery strategy for gene therapy of perioperative myocardial ischemia.