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目的:探讨在心肌缺血/再灌注和腺苷诱导预适应过程中细胞膜钙泵(PMCA)的可能作用。方法:将培养的乳鼠心肌细胞分为3组:缺血/再灌注组、腺苷预处理组和对照组。各组乳酸脱氢酶(LDH)漏出量以生化法检测(n=5);半定量RT-PCR检测各组PMCA基因转录水平(n=3);PMCA活性以定磷比色法测定(n=5)。结果:(1)缺血/再灌注组LDH漏出量明显增高(P<0.001),腺苷预处理组LDH漏出量显著降低(P<0.001);(2)各组间PMCA mRNA转录水平无显著性差异(P>0.05);(3)各组间PMCA蛋白活性无显著性差异(P>0.05)。结论:在乳鼠心肌细胞缺血/再灌注过程中PMCA无代偿功能;PMCA不直接参与腺苷诱导预适应的心肌保护作用。
Objective: To investigate the possible role of PMCA during myocardial ischemia / reperfusion and adenosine-induced preconditioning. Methods: The cultured neonatal rat cardiomyocytes were divided into 3 groups: ischemia / reperfusion group, adenosine preconditioning group and control group. The leakage of lactate dehydrogenase (LDH) in each group was detected by biochemical method (n = 5). The transcription level of PMCA gene in each group was detected by semi-quantitative RT-PCR (n = 3) = 5). Results: (1) LDH leakage in ischemia / reperfusion group was significantly increased (P <0.001), LDH leakage in adenosine preconditioning group was significantly decreased (P <0.001); (2) PMCA mRNA transcript level was not significant (P> 0.05). (3) There was no significant difference in PMCA protein activity between groups (P> 0.05). CONCLUSION: PMCA has no compensatory function in neonatal rat cardiomyocytes during ischemia / reperfusion. PMCA is not directly involved in myocardial protection induced by adenosine preconditioning.