Expression Of AEs And CLCs Protein On Rat's Cardiocytes Suffered From Chloridion-free/re-chloridion Injury

Objective: To observe whether cardiocytes were damaged after the treatment of chloridion-free/re-chloridion. and to investigate the expression of AEs and CLCs protein on the cardiocytes sufferd from chloridion-free/re-chloridion injury. Methods: Cultured cardiocytes of neonatal SD rats had been randomly divided into two groups: Control group, and Chloridion-free/re-chloridion group. At the end of experiment, cardiocytes beating rates, cell viabilities and the activities of LDH in culture were measured, furthermore. The morphology assessment of cardiocytes was observed by inverted microscope, the expression of AEs and CLCs mRNA were measured by RT-PCR and the expression of AEs protein were measured by western blotting. Results: The beating rates and the cell viabilities were significantly difference between chloridion-free/re-chloridion group and control group(p<0.01), and the cell viabilitis of chloridion-free/re-chloridion was significantly decreased(p<0.01), and LDH activities of chloridion-free/re-chloridion group was significantly increased(p<0.01). AE1, AE2, AE3, CLC-2, CLC-3, CLC-4, CLC-5, CLC-6, CLC-7 mRNA was revealed by RT-PCR that showed a basic expression in control group. CLC-2, CLC-4 and AE2 mRNA were similar to the two group(p>0.05). AE1 and AE3 mRNA had significantly uprelated (p<0.01), and CLC-3, CLC-5, CLC-6, CLC-7 mRNA expression had significantly downrelated(p<0.01) of cardiocytes in chloridion-free/re-chloridion group compared with control group. Western blotting revealed all of AE1, AE2, and AE3 protein had a basic expression in control group. The diversity tendency of AE1, AE2, AE3 protein were the same as the mRNA of those. AE1 and AE3 protein expression in chloridion-free/re-chloridion group had a significantly uprelated ,and AE2 protein was similar to that of control group(p>0.05). Conclusion: It is verified that the chloiridion-free/re-chloridion could lead to myocardial injury, moreover, AEs and CLCs protein potentially participate the myocytes chloridion-free/re-chloridion injury.