N-乙酰半胱氨酸对培养心肌细胞缺氧-复氧性损伤的干预研究

Study of the effect of N-acetyl-L-cysteine on culture myocardial cells with hypoxia reoxygenation injury in rats

目的 :探讨缺氧复氧性损伤的发生机制及 N乙酰半胱氨酸 (NAC)对损伤的影响。方法 :Wistar大鼠 5 0只 ,取其心肌细胞和主动脉内皮细胞培养 ,分离其外周血中性粒细胞 ,分设正常条件培养组 ,单纯缺氧复氧组 (H R) ,正常培养 +白介素 1β(IL 1β) 1× 10 5/ L 刺激组 ,将以上 3组再分别分为未干预组和 NAC(80 m g/ L)干预组。于缺氧 1小时后复氧 6小时 ,测定心肌细胞 IL 1β、细胞间黏附分子 1(ICAM 1)蛋白质表达水平、细胞胞浆游离钙 (〔 Ca2 + 〕i)、一氧化氮合酶 (NOS)、超氧化物歧化酶 (SOD)、丙二醛 (MDA)、还原型谷胱苷肽 (GSH)和中性粒细胞与内皮细胞黏附率。结果 :心肌细胞 H R及 IL 1β刺激时 ,IL 1β、ICAM 1蛋白质表达 ,中性粒细胞与内皮细胞的黏附率、〔 Ca2 + 〕i和 MDA均有显著增加 ,而 SOD、GSH、NOS明显下降 ;但H R组上述各项指标除 GSH外 ,改变较 IL 1β干预组更显著。 NAC可明显改善 2组 IL 1β、ICAM 1的表达和 MDA、〔 Ca2 +〕i超载增高的程度以及 SOD、GSH、NOS下降程度 ,降低中性粒细胞与内皮细胞的黏附率(P<0 .0 5或 P<0 .0 1)。结论 :IL 1β可通过直接或间接激活 ICAM 1导致中性粒细胞与内皮细胞黏附增加 ,引起细胞损伤 ,但钙超载、氧自由基和 NO NOS系统也共同参与了缺氧复氧性损?

Objective:To investigate the mechanism of hypoxiareoxygenation (HR) injury to myocardial cells in culture and to evaluate the effects of NacetylLcysteine (NAC) on the injury.Methods:Fifty rats were divided into three groups:HR group,interleukin1β(IL1β) group and control group.Animals in each group were divided equally into intervention with IL1β Ab and non intervention groups.Samples were observed at 1 hour after hypoxia and 6 hours after reoxygenation.The expressions of IL1β and intercellular adhesion molecule1(ICAM1) protein level were measured by enzyme linked immunoadsorbent assay (ELISA) technique.The adhesive rate of polymorphonuclear leukocytes(PMNs) to endothelial cells (ECs) were observed ,and the content of malondialdehyde(MDA),superoxide dismutase(SOD),reduced glutathione (GSH), nitric oxide synthase(NOS),〔Ca 2+ 〕i,concentration in myocardial cells were also measured. Results: The expression of ICAM1,IL1β protein,〔Ca 2+ 〕i,MDA and adhesive rate of PMNsECs were increased and SOD,GSH,NOS were decreased at 6 hours after HR.In IL1β group,similar changes were observed. All these changes could be attenuated by NAC( P <0 05 or P <0 01).Conclusions:HR injury to myocardial cells is a complex of consequences resulted from multiple factors.The injury can be enhanced by inducing changes in SOD,GSH,NOS,〔Ca 2+ 〕i,expression of ICAM1,and adhesive rate of PMNsECs effected by IL1β. This injury can be partly attenuated by NAC.