培养心肌细胞缺氧复氧性损伤机制及ICAM-1 MAb的干预研究

Ths study of the mechanisms of culture myocardial cells hypoxia reoxygenation injury in rats and the effect of ICAM\|1MAb

目的 :探讨缺氧复氧性损伤的发生机制 ,澄清IL 1在损伤中的作用及ICAM 1MAb对损伤的影响。方法 :新生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 (HR) ,正常培养 +IL 1β(10 0u ml)刺激组 ,将以上 3组再分别分为 :不干预组和ICAM 1MAb(10 0ng ml)干预组 ;于缺氧 1h复氧 6h ,测定心肌细胞一氧化氮合酶 (NOS) ,细胞胞浆游离钙 ([Ca2 + ]i) ,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粘附率。结果 :心肌细胞缺氧复氧时 ,[Ca2 + ]i、MDA显著增加 ,而SOD、GSH、NOS明显下降 ;IL 1β、ICAM 1蛋白质表达 ,中性粒细胞与内皮细胞的粘附率显著增加。IL 1干预刺激组的上述指标变化类似于缺氧复氧组。ICAM 1MAb可明显改善SOD、GSH及NOS下降的程度和MDA增高的程度 ,降低中性粒细胞与内皮细胞的粘附率 ,但不能改善 [Ca2 + ]i超载和IL 1β的表达水平 ,P >0 0 5。 结论 :IL 1β通过直接或激活ICAM 1导致中性粒细胞与内皮细胞的粘附增加而引起细胞损伤 ,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伤。ICAM 1MAb通过抑制中性粒细胞系统、减轻氧自由基产生、改善NOS代谢可部分减轻损伤。

Objective:To investigate the mechanism of culture myocardial cells hypoxia/reoxygenation(HR) and to evaluate the effects of intercellular adhesion molecule\|lmonoclonal antibody(ICAM\|1 MAb) to the damage.Method:50 rats were divided into three groups:HR group,IL\|1β group and control group.Then,animals in each groups were divided equally into interventions of ICAM\|1 MAb and non\|interventions groups.Samples were observed at the time of 1h after hypoxia and 6h after reoxygenation.The expression of ICAM\|1 and IL\|1β protein level were measured by ELISA technique.The adhesive rate of polymorphonuclear leukocytes(PMNs) on to endothelial cells (ECs),SOD,GSH in myocardial cells,nitric oxide synthase (NOS),[Ca~ 2+]i concentration were also observed.Result:The expression of ICAM\|1,IL\|1β protein,[Ca~ 2+]i,MDA and adhesive rate of PMNs\|ECs were increased and SOD,GSH,NOS were decreased at the time of 6h after HR.In IL\|1β groups,similar changes were observed.The changes of SOD,GSH,NOS,adhesive rate of PMNs\|NCs can be attenuate by ICAM\|1 MAb,but the expression of ICAM\|1,IL\|1β protein and [Ca~ 2+]i cannot be affected. P >0.05.Conclusion:Myocardial cells HR injury was a complex consequences resulted from multiple factors,The injury can be enhanced by induced the changes of SOD,GSH,NOS,[Ca~ 2+]i,expression of ICAM\|1,and adhesive rate of PMNs\|Ecs by IL\|1β.This injury can be partly reduced by attenuate these changes by ICAM\|1 Mab.