Antiapoptotic effects of rat hypertrophic cardiomyocytes induced with hypoxia/reoxygenation by dichloroacetate

Objective To investigate the effect of hypoxia/reoxygenation on apoptosis of rat hypertrophic cardiomyocytes and the metabolic pathway changes of hypertrophic cardiomyocytes by dichloroacetate.Methods The isolated ventricular cardiomyocytes from neonatal Wistar rat were cultured and purified with differential attachment,then BrdU was added to reduce the rate of non-myocytes.The cultured cells were identified by morphology,spontaneous contraction and specific immunocytochemical stain.Hypertrophic cardiomyocyte model induced by angiotensin Ⅱ was set up and assessed by -Leu incorporation.The cultured media were replaced by low glucose DMEM before hypoxia.The cardiomyocytes were incubated at 37 ℃ in an air-tight incubator containing 92% N_(2),5% CO_(2),3% O_(2) for 24 h to simulate hypoxia,then under the condition of 23% O_(2),5% CO_(2) for 4 h to simulate reoxygenation.Apoptotic cells was evaluated by a modified TUNEL assay(DeadEnd~(TM) Colorimetric TUNEL).Results The apoptotic rate of cardiomyocytes increased with time prolongation of hypoxia/reoxygenation(H/R).The apoptotic cell model induced by H/R in rat hypertropic cardiomyocytes was established successfully.When the hypertropic cardiomyocytes were pretreated with Dichloroacetate,the apoptotic rate decreased in a dose-dependent manner.The metabolic pathway changes had effect on the apoptosis of rat hypertrophic cardiomyocytes.Conclusion By collagense digestion,it is convenient to obtain primary neonatal Wistar rat ventricular cardiomyocytes. This method is one of vital research tools in the fields of cardiac hypertrophy.Angiotensin Ⅱ could accelerate cellular proliferation,structural protein biosynthesis and cardiac hypertrophy.Dichloroacetate has the effect of antiapoptosis.