一氧化氮对培养乳鼠心肌细胞低氧复氧损伤的影响

Effect of nitric oxide on anoxia/reoxygenation injuries of neonatal rat cardiomyocytes

目的 :研究 NO对乳鼠心肌细胞低氧 /复氧 (A/R)损伤的影响。方法 :建立心肌细胞 A/R损伤模型 ,随机分为6组 :A组 ,正常对照组 ;B组 ,单纯低氧 /复氧 (A/R低氧 2 h,复氧 1h) ;C组 ,低氧预处理 ,低氧 2 0 m in后复氧 2 0min,然后 A/R;D、E、F组为 NO预处理组 ,加入 S-亚硝基 -已酰青酶胺 (SNAP)分别使其终浓度为 0 .5、1、2 mm ol/L,预处理 4 0 min后 A/R。于复氧后测定各组培养液中肌酸激酶 (CK)、蛋白漏出量的变化及细胞内丙二醛 (MDA)含量和细胞存活率。结果 :单纯低氧 /复氧组和 2 m mol/L SNAP组 CK、蛋白漏出量、MDA水平显著升高 (P<0 .0 1vs正常组 ) ,细胞存活率显著降低 (P<0 .0 1vs正常组 )。0 .5和 1mmol/L SNAP预处理组和低氧预处理组上述变化明显减轻 (P<0 .0 1vs低氧复氧组 )。结论 :NO预处理对乳鼠心肌细胞 A/R损伤具有保护作用 ,并具有浓度依赖性。

AIM: To study the effect of Nitric oxide(NO) on anoxia/reoxygenation (A/R) injuries of neonatal rat cardiomyocytes. METHODS: An anoxia/reoxygenation model of myocardial cells of neonatal SD rats was established. The cells were randomly divided into six groups: control group (A): without any pretreatment; anoxia/reoxygenation (A/R) group (B): 2-hour anoxia followed by 1-hour reoxygenation; anoxia preconditioning group (C): 20- minute anoxia followed by 20-minute reoxygenation and then A/R; NO preconditioned group (D): adding s-nitroso-n-acetyl-penicillamine(SNAP) into culture medium till the final concentration of 0.5 mmol/L, after 40 minutes preconditioning and then A/R; NO preconditioned group (E): adding SNAP into culture medium till the final concentration of 1 mmol/L, after 40 minutes preconditioning and then A/R; NO preconditioned group (F): adding SNAP into culture medium till the final concentration of 2 mmol/L, after 40 minutes' preconditioning and then A/R. The activity of creatine kinase (CK), the leakage of protein, the contents of cellular malondiadehyde (MDA), the rate of cell viability of each group were evaluated after reoxygenation. RESULTS: The anoxia/reoxygenation group and 2 mmol/L SNAP group showed a great increases in levels of CK,protein and MDA, and a decrease in cell viability(P<0.01 vs group A). The anoxia preconditioned,the 0.5 mmol/L and the 1 mmol/L SNAP groups significantly attenuated these change(P<0.01 vs group B). CONCLUSION: NO can exert a protective effect on A/R injury of neonatal rat cardiomyocytes in a concentration dependent manner.