兔心肌缺血再灌注损伤中内皮细胞功能变化与参麦复方的干预效应(英文)

Effect of shenmai injection on endothelial function during myocardial ischemia/reperfusion injury in rabbits

背景:一氧化氮和内皮素是近年心肌缺血再灌注损伤研究的热点。中药在保护心肌缺血再灌注损伤中有其独特的作用。目的:观察兔心肌缺血再灌注损伤中内皮功能改变及参麦注射液的干预作用。设计:随机对照动物实验。单位:中国医科大学附属第一医院。材料:实验于2002-09/2003-01在中国医科大学实验动物部、基础医学院生化教研室、第二电镜室完成。健康雌性日本大耳白兔21只,体质量1.5～2.5kg;参麦注射液由雅安三九药业有限公司提供(010110)。方法:①分组:实验动物随机分3组,每组7只。假手术对照组:只穿线不结扎冠状动脉。心肌缺血再灌注模型组和心肌缺血再灌注+参麦注射液治疗组:冠状动脉左室支结扎40min后放开40min建立心肌缺血再灌注模型。②给药:心肌缺血再灌注模型组耳缘静脉注射生理盐水20mL(于左室支结扎前10min注射1/3量,再灌注时注射2/3量)。心肌缺血再灌注+参麦注射液治疗组,耳缘静脉注射参麦注射液(按1.5mL/kg,用生理盐水稀释至20mL),给药时间同心肌缺血再灌注模型组。③观察指标:于结扎前、缺血40min、再灌注40min取静脉血4mL,其中2mL分离血清采用硝酸还原酶法检测一氧化氮代谢产物含量;另2mL采用放免法检测血浆内皮素含量。于实验结束后,取心肌组织检测一氧化氮代谢产物、内皮素,采用黄嘌呤氧化酶法检测总超氧化物歧化酶活性,采用硫代巴比妥酸显色法检测丙二醛含量。透射电镜观察心肌超微结构。主要观察指标:①结扎前、缺血40min、再灌注40min血清一氧化氮、血浆内皮素的含量。②心肌组织一氧化氮代谢产物、内皮素、丙二醛的含量和总超氧化物歧化酶活性。③心肌超微结构。结果:21只日本大耳白兔均进入结果分析。①血清一氧化氮代谢产物含量:缺血40min、再灌注40min心肌缺血再灌注模型组明显低于假手术对照组(P<0.01),心肌缺血再灌注+参麦注射液治疗组显著高于心肌缺血再灌注模型组(P<0.01)。②血浆内皮素含量:缺血40min、再灌注40min心肌缺血再灌注模型组显著高于假手术对照组(P<0.01),心肌缺血再灌注+参麦注射液治疗组明显低于心肌缺血再灌注模型组(P<0.01)。③缺血40min、再灌注40min一氧化氮代谢产物含量与内皮素含量呈显著负相关(P<0.05,P<0.01)。④心肌组织一氧化氮代谢产物含量、总超氧化物歧化酶活性:再灌注40min后心肌缺血再灌注模型组明显低于假手术对照组(P<0.01),心肌缺血再灌注+参麦注射液治疗组显著高于心肌缺血再灌注模型组(P<0.01)。⑤内皮素,丙二醛含量:心肌缺血再灌注模型组显著高于假手术对照组(P<0.01),心肌缺血再灌注+参麦注射液治疗组明显低于心肌缺血再灌注模型组(P<0.01)。⑥一氧化氮代谢产物含量与总超氧化物歧化酶活性呈显著正相关(P<0.05),与丙二醛含量呈显著负相关(P<0.05),内皮素含量与总超氧化物歧化酶活性呈显著负相关(P<0.05),与丙二醛含量呈显著正相关(P<0.05)。结论:心肌缺血再灌注导致血管内皮功能紊乱;参麦注射液能够改善缺血再灌注时的内皮功能,减轻心肌缺血再灌注损伤。

BACKGROUND： With the extensive development of reperfusion treatment on acute myocardial infarction, myocardial ischemia/reperfusion （I/R） injury cause clinical concern intensively. In recent years, nitric oxide （NO） and endothelin （ET） are hot point of researches. Myocardial protective role of Chinese traditional medicine is highly regarded. OBJECTIVE ： To study the change of endothelial function during myocardial I/R injury of rabbits, the effect of shenmai injection （SMI） and mechanism. DESIGN ： Randomized controlled animal study SETTING： The First Hospital Affiliated to China Medical University MATERIALS： The experiment was carried out in Laboratory Animal Center, Department of Biochemistry and the Second Electron Microscope Room of China Medical University from September 2002 to January 2003. Twenty-one healthy female Japanese white rabbits weighing 1.5-2.5 kg were in this study. SMI was produced by the Three Nine Medicine Industry Co., Ltd., Yaan （batch number： 010110）. METHODS： （1） Grouping： The animals were randomly divided into three groups, 7 cases of each group. Sham-operation group： Rabbits were gone through thread only and were not ligated coronary artery; I/R group and I/R ＋ SMI group： Coronary artery occlusion was maintained 40 minutes at the ligature was released and reperfusion continued for 40 minutes. （2） Administration： Rabbits in I/R group were injected 20 ml saline through ear margin vein （injected one third of saline at 10 minutes before ligature, two thirds of saline when unloosing the ligature）. Rabbits in I/R ＋ SMI group were injected with SMI which was diluted with saline to 20 mL （1.5mL/kg） Index observation： 4 mL venous blood samples were 5 mL/kg）. The method of injection was as same as I/R group. obtained before ligature, at 40 minutes of ischemia and 40 minutes of reperfusion. Half of the blood samples were separated serum to detect NO production （NOP） by nitric acid deacidizing enzyme process; another were separated plasma to detect ET by radiation immunity process. In the experiment ends, NOP and ET of myocardial tissue were detected; total of superoxide dismutase （T-SOD） of myocardial tissue were detected by sulphur purine oxidase process and malondialdehyde （MDA） were detected by thiobarbituric acid development process. Myocardial ultrastructure was observed with transmission electro microscope. MAIN OUTCOME MEASURES： （1） Contents of serum NO and plasma ET in each group before ligature, at 40 minutes of ischemia and 40 minutes of reperfusion; （2） contents of NOP, ET, T-SOD and MDA of myocardial tissue; （3） Myocardial ultrastructure. RESULTS ： All 21 rabbits were involved in the final analysis. （1） Content of serum NOP： Content of serum NOP was lower in I/R group than that in sham-operation group at 40-minute ischemia and 40-minute perfusion （P 〈 0.01 ）, but was higher in I/R ＋SMI group than that in I/R group （P 〈 0.01）. （2） Content of plasma ET： Content of plasma ET was higher in I/R group than that in sham-operation group at 40-minute ischemia and 40-minute perfusion （P 〈 0.01）, but was lower in I/R ＋SMI group than that in I/R group （P 〈 0,01）. （3） Content of NOP was negative correlation with that of ET 40-minute ischemia and 40-minute perfusion （P 〈 0.05, P 〈 0.01）. （4） Content of NOP and activity of T-SOD： Those were lower in I/R group than those in sham-operation group at 40-minute perfusion （P 〈 0.01）, but were higher in I/R ＋SMI group than those in I/R group （P 〈 0.01）. （5） Contents of ET and MDA： Those were higher in I/R group than those in sham-operation group （P 〈 0.01 ）, but were lower in I/R ＋SMI group than those in I/R group （P 〈 0.01 ）. （6） Content of NOP was positive correlation with T-SOD activity （P 〈 0.05）, and negative correlation with content of MDA （P 〈 0.05）, In addition, content of ET was negative correlation with T-SOD activity （P 〈 0.05）, and positive correlation with content of MDA （P 〈 0.05）. CONCLUSION： Myocardial I/R leads to abnormality of vascular endothelial function, and SMI can improve endotheliual function and relieve myocardial I/R injury.