摘要
目的研究一氧化氮供体S-亚硝基-已酰青酶胺(SNAP)对缺氧/复氧乳鼠心肌细胞内游离钙浓度的影响.方法培养20×5只SD大鼠(1～3)d的心肌细胞,随机分为5组:A组为正常对照组;B组为单纯缺氧/复氧组(A/R缺氧2 h复氧1 h);C组为NO预处理组加入SNAP使其终浓度分别为0.1 mmol/L,预处理20 min后行A/R;D组为NO预处理组加入SNAP使其终浓度为1 mmol/L,预处理20 min后行A/R;E组为NO预处理组加入SNAP使其终浓度为2 mmol/L,预处理20 min后行A/R.与复氧后应用特异性Ca2+荧光指示剂Fluo-3/AM负载细胞,激光共聚焦显微镜检测游离Ca2+浓度.结果与正常组相比,单纯缺氧/复氧组钙浓度明显升高(P<0.01);0.1 mmol/L和1 mmol/L SNAP能明显降低钙超载(与B组相比,P<0.01),2 mmol/L SNAP组加重钙超载(与B组相比,P＞0.05).结论一氧化氮通过降低细胞内钙超载而减轻缺氧/复氧对心肌细胞的损伤,其作用有浓度依赖性.
Objective To study the effects of nitric oxide donor （SNAP） on intracellular calcium in anoxia/ reoxygenation injury of neonatal rat cardiomyocytes. Methods The cultured myocardial cells of neonatal SD rats were randomly divided into five groups, control group （A）： without any treatment; anoxia/reoxygenation group （B）： 2h anoxia followed by l h reoxygenation; NO preconditioning groups （C）： adding SNAP into culture medium to form different final concentrations, for 20 minutes preconditioning and then A/R. After reoxygenation, intracellular free Ca^2＋ concentration was measured by calcium fluorescent probe Fluo3/AM and laser confocal microscope. Results Compared with control group, anoxia/reoxygenation group caused great overload of intracellular Calcium （ P 〈 0.01）. 0.1 mmol/L SNAP and lmmol/L SNAP significantly attenuated the change （ P 〈 0.01 vs B）, and 2 mmol/L SNAP group aggravated the level of high intracellular Calcium. Conclusions NO alleviated the anoxia/reoxygenation injury on neonatal rat cardiomyocytes by attenuating the overloading of intracellular calcium, and the effects were concentration-dependent.
出处
《中华急诊医学杂志》
CAS
CSCD
2005年第12期1007-1009,共3页
Chinese Journal of Emergency Medicine
