地高辛抗血清对老龄大鼠心肌缺氧复氧损伤影响的体外实验  

In vitro experiment of anti-digoxin antiserum effect on myocardial hypoxia reoxygenation injury in aging rats

作　　者：王德国[1] 杨玉雯[1] 陈月云[1] 唐振国[1] 

机构地区：[1]皖南医学院附属弋矶山医院老年心血管科,安徽省芜湖市241001

出　　处：《中国组织工程研究与临床康复》2007年第8期1535-1537,共3页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：皖南医学院中青年科研基金(WK200421A)~~

摘　　要：目的:观察内洋地黄素水平在老龄大鼠心肌细胞缺氧复氧损伤中的变化以及地高辛抗血清的保护作用。方法:实验于2005-04/05在皖南医学院病理生理教研室完成。取10只24月龄和10只6月龄雄性普通级SD大鼠,分别制备青年和老龄大鼠心肌细胞匀浆,老龄和成年大鼠为两大组,每组分为7小组,即每只大鼠心肌随机分到各小组中,共计14小组,每组10支试管。正常对照组:给予CO2和O2的混合气体(1∶19)通气40min;缺氧复氧组:CO2,O2,N2混合气体(5∶4∶91)通气20min后换成CO2和O2的混合气体(1∶19)通气20min;阴性对照组:同缺氧复氧组,但于再给氧前加入0.1mL的非特异性灭活兔血清;地高辛抗血清组:同缺氧复氧组,但于再给氧前加入0.1mL的非特异性地高辛抗血清(分别为1∶90000,60000,30000,10000)。观察大鼠心肌细胞钠-钾-三磷酸腺苷酶活性和线粒体内钙聚集程度,分析其剂量-效应关系。结果:①缺氧复氧时,青年组和老龄组大鼠心肌分泌内洋地黄素均显著升高,但老龄组显著低于青年组[(0.081±0.03),(0.153±0.06);(0.074±0.04),(0.125±0.05)ng/g;P<0.05]。②缺氧复氧时,老龄组与青年组心肌细胞钠-钾-三磷酸腺苷酶活性显著受抑制[(0.239±0.015),(0.778±0.050);(0.350±0.047),(0.836±0.044)μkat/g;P<0.05],老龄组与青年组相比,其抑制效应显著增强(P<0.05)。③缺氧复氧时,老龄组线粒体内钙与青年组比较明显增强[(0.082±0.011),(0.495±0.095);(0.075±0.008),(0.412±0.084)mmol/L,P<0.05]。④老龄组和青年组相比,地高辛抗血清呈剂量依赖性的恢复钠-钾-三磷酸腺苷酶活性(r=0.695,0.797,n=5,P<0.05),减轻线粒体内钙聚集(r=-0.565,-0.649,n=5,P<0.05);经直线回归分析发现,老龄鼠回归系数大于青年组(酶活性抑制K=1.50,0.94,线粒体内钙K=-7.43,-6.46)。结论:心肌细胞缺氧复氧时,老龄鼠损伤较青年大鼠更显著,其机制与老龄大鼠心肌细胞钠-钾-三磷酸腺苷酶对内洋地黄素敏�AIM： To investigate the change of endoxin in myocardial hypoxia reoxyenation injury in aging rats and the protective effect of digoxin antiserum. METHODS： The experiment was performed at the Department of Pathophysiology in Wannan Medical College from April to May 2005. Totally 10 24-month and 10 6-month male SD rats of common grade were selected to establish myocardial cell homogenate models of youth and ageing rats, and divided into aging group and adult group, respectively with 7 subgroups in each group. Myocardium of each rat was assigned into each subgroup, totally 14 subgroups with 10 tubes in each group. Normal control group received the mixed gas of CO2 and O2 （1：19） for 40 minutes; hypoxia reoxyenaUon group received mixed gas of CO2, O2 and N2 （5：4：91） for 20 minutes, and then changed into the mixed gas of CO2 and O2 （1：19） for 20 minutes; negative control group received the same process as that in the hypoxia reoxyenation group, but intervened with 0.1 mL non-specific inactive rabbit serum before giving oxygen; digoxin antiserum group： the same as the hypoxia reoxyenation group, but 0.1 mL non-specific anti-digoxin antiserum of different concentration （1：90 000, 60 000, 30 000 and 10 000, respectively） was given before giving oxygen. Activity of Na^＋-K^＋-ATPase and Ca^2＋ accumulation in mitochondria were observed in myocardial cells of rats and the doseeffect relationship was analyzed. RESULTS： ①During hypoxia reoxyenation, endoxin level of myocardium remarkably increased in the youth group and aging group, but it was lower significantly in the aging group than the youth group [（0.081±0.03）, （0.153±0.06） ; （0.074± 0.04）, （0.125±0.05） ng/g;P 〈 0.05].②During hypoxia reoxyenaUon, the activity of Na^＋-K^＋-ATPase was inhibited remarkably in the aging group and youth group [（0.239±0.015）, （0.778±0.050）;（0.350±0.047）, （0.836±0.044） μkat/g： P 〈 0.05]. The inhibitive effect in the aging group increased obviously as

关 键 词：洋地黄糖甙类/代谢 地高辛/药理学 缺氧/病理生理学 钙/代谢 

分 类 号：R363[医药卫生—病理学]