| Objective: To study the role and mechanism of AE family in anoxic preconditioning of primary culture neonatal rat cadiocyte. Methods: An anoxic preconditioning model was established using cultured neonatal rat cardiomyocytes, Pharmacological agents (L-NAME 50 μM, glibenclamide 50 μM and PD98059 50 μM) were used to inhibit NO, K+ATP channel and MAPK channel activation during anoxic preconditioning. Cultured myocardial cells of neonatal SD rats have been randomly divided into six groups: Control group; anoxia reoxygenation (A/R) group;delayed preconditioning(DPC) group; PD98058+DPC group; glibenclamide +DPC group; L-NAME+DPCgroup. Cellular injury was evaluated by measuring myocardial cells beating rate, cell viability, and lactate dehydrogenase (LDH) release. The expression of AEs mRNA and protein were measured by RT-PCR and western blotting, respectively. Results: 1. Beating rate of ventricular myocytes: The beating rate of ventricular myocytes in A/R group(15±7 beats/min) was significantly slow compared with that in control group (p<0.01) ; Compared with A/R group, The beating rate of ventricular myocytes in DPC group(87±10 beats/min) significantly increased (p<0.01) ; The beating rate of PD98059+DPC group(16±6 beats/min) , Glibenclamide+DPC group(18±7 beats/min), L-NAME+DPC group (14±5 beats/min) was significantly lower than DPC group(p<0.01) . 2. the cell viability: The cell viability of A/R(38.9±7.2%) group significantly decreased when compared with that of control group (p<0.01); The viability of DPC (78.2±10.6%) group significantly increased compared with that of A/R group respectively (p<0.01) ; There was no significant difference in viability of the three groups respectively, PD98059+DPC group (40.3±6.2%), Glibenclamide+DPC group ( 38.9±7.4% ) and L-NAME+DPC group(40.6±8.7% ) Compared with that of A/R group (p>0.05) . While the difference between that of DPC group and that of the three groups was significant...
|
PDF Full Text Request