Abstract: Objective: To investigate the effect of pirarubicin (THP) on the cardiomyocyte injury in the rats, and to analyze the establishment method of THP-induced cardiomyocyte injury model. Methods: The rat cardiomyocytes H9C2 were routinely cultured in vitro. The cardiomyocytes H9C2 were divided into blank control group(0 μmol·L^-1 group) and different concentrations(1×10^-6 mol·L^-1, 1×10^-5 mol·L^-1, 1×10^-4 mol·L^-1, 1×10^-3 mol·L^-1, 1 μmol·L^-1, 3 μmol·L^-1, 5 μmol·L^-1, 7 μmol·L^-1, 9 μmol·L^-1,and 10 μmol·L^-1)of TNP groups. The H9C2 cells were treated with different concentrations of THP; at 6, 12, 24, 36 and 48 h, the survival rates of cells in various groups were detected by CCK-8 method; DCFH-DA reactive oxygen species (ROS) probe method was used to detect the intracellular ROS level; the malonaldehyde (MDA) levels in the cells in various groups were detected by thiobarbituric acid colorimetry, the superoxide dismntase(SOD) activities in the cells in various groups were detected by xanthine oxidation, and the lactic dehydrogenase(LDH) activities in the cells in various groups were detected by dinitrophenylhydrazine colorimetry; the apoptotic rates in various groups were determined by TUNEL staining; the expression levels of Bax, Bcl-2, Caspase-3 and Caspase-9 mRNA in the H9C2 cells in various groups were determined by qRT-PCR method;the expression levels of Bax, Bcl-2, Cleaved Caspase-3, and Cleaved Caspase-9 proteins in H9C2 cells in various groups were determined by Western blotting method. Results: Compared with blank control group, the survival rates of H9C2 cells in 1, 5 and 9 μmol·L^-1 THP groups were decreased (P<0.05 or P<0.01), the intracellular ROS and MDA levels and LDH activities were increased (P<0.05 or P<0.01), the SOD activities were decreased (P<0.05 or P<0.01), the apoptotic rates were increased(P<0.05), the expression levels of Bax, Caspase-3 and Caspase-9 mRNA were increased(P<0.05 or P<0.01), and the expression levels of Bcl-2 mRNA were decreased (P<0.01);the expression levels of Bax, Cleaved Caspase-3 and Cleaved Caspase-9 proteins were increased(P<0.05 or P<0.01), and the expression levels of Bcl-2 protein were decreased (P<0.01). Conclusion: THP has a damaging effect on the rat cardiomyocytes, of which 5 μmol·L^-1 is the optimal modeling concentration of THP-induced H9C2 injury model.

Key words: pirarubicin, cardiomyocytes, disease model,animal, apoptosis, cell injury