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
토심기결혈재관주손상중내피세포공능변화여삼맥복방적간예효응
Effect of shenmai injection on endothelial function during myocardial ischemia/reperfusion injury in rabbits
  • 万方数据
    中国组织工程研究与临床康复 중국조직공정연구여림상강복
    JOURNAL OF CLINICAL REHABILITATIVE TISSUE ENGINEERING RESEARCH
    2007年 21期 4244-4247 ,共4页

    郭莲怡%金旭鹏%刘沛

    곽련이%금욱붕%류패

    心肌缺血%再灌注损伤%一氧化氮%内皮缩血管肽%参麦注射液 心肌缺血%再灌註損傷%一氧化氮%內皮縮血管肽%參麥註射液
    심기결혈%재관주손상%일양화담%내피축혈관태%삼맥주사액
    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1.5～2.5 kg;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40 min后放开40 min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20 mL(于左室支结扎前10 min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1.5 mL/kg,用生理盐水稀释至20 mL),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40 min、再灌注40 min取静脉血4 mL,其中2 mL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2 mL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40 min、再灌注40 min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40 min、再灌注40 min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40 min、再灌注40 min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40 min、再灌注40 min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40 min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.
    배경:일양화담화내피소시근년심기결혈재관주손상연구적열점.중약재보호심기결혈재관주손상중유기독특적작용.목적:관찰토심기결혈재관주손상중내피공능개변급삼맥주사액적간예작용.설계:수궤대조동물실험.단위:중국의과대학부속제일의원.재료:실험우2002-09/2003-01재중국의과대학실험동물부、기출의학원생화교연실、제이전경실완성.건강자성일본대이백토21지,체질량1.5～2.5 kg;삼맥주사액유아안삼구약업유한공사제공(010110).방법:①분조:실험동물수궤분3조,매조7지.가수술대조조:지천선불결찰관상동맥.심기결혈재관주모형조화심기결혈재관주+삼맥주사액치료조:관상동맥좌실지결찰40 min후방개40 min건립심기결혈재관주모형.②급약:심기결혈재관주모형조이연정맥주사생리염수20 mL(우좌실지결찰전10 min주사1/3량,재관주시주사2/3량).심기결혈재관주+삼맥주사액치료조,이연정맥주사삼맥주사액(안1.5 mL/kg,용생리염수희석지20 mL),급약시간동심기결혈재관주모형조.③관찰지표:우결찰전、결혈40 min、재관주40 min취정맥혈4 mL,기중2 mL분리혈청채용초산환원매법검측일양화담대사산물함량;령2 mL채용방면법검측혈장내피소함량.우실험결속후,취심기조직검측일양화담대사산물、내피소,채용황표령양화매법검측총초양화물기화매활성,채용류대파비타산현색법검측병이철함량.투사전경관찰심기초미결구.주요관찰지표:①결찰전、결혈40 min、재관주40 min혈청일양화담、혈장내피소적함량.②심기조직일양화담대사산물、내피소、병이철적함량화총초양화물기화매활성.③심기초미결구.결과:21지일본대이백토균진입결과분석.①혈청일양화담대사산물함량:결혈40 min、재관주40 min심기결혈재관주모형조명현저우가수술대조조(P＜0.01),심기결혈재관주+삼맥주사액치료조현저고우심기결혈재관주모형조(P＜0.01).②혈장내피소함량:결혈40 min、재관주40 min심기결혈재관주모형조현저고우가수술대조조(P＜0.01),심기결혈재관주+삼맥주사액치료조명현저우심기결혈재관주모형조(P＜0.01).③결혈40 min、재관주40 min일양화담대사산물함량여내피소함량정현저부상관(P＜0 05,P＜0.01).④심기조직일양화담대사산물함량、총초양화물기화매활성:재관주40 min후심기결혈재관주모형조명현저우가수술대조조(P＜0.01),심기결혈재관주+삼맥주사액치료조현저고우심기결혈재관주모형조(P＜0.01).⑤내피소,병이철함량:심기결혈재관주모형조현저고우가수술대조조(P＜0.01),심기결혈재관주+삼맥주사액치료조명현저우심기결혈재관주모형조(P＜0.01).⑥일양화담대사산물함량여총초양화물기화매활성정현저정상관(P＜0.05),여병이철함량정현저부상관(P＜0.05),내피소함량여총초양화물기화매활성정현저부상관(P＜0.05),여병이철함량정현저정상관(P＜0.05).결론:심기결혈재관주도치혈관내피공능문란;삼맥주사액능구개선결혈재관주시적내피공능,감경심기결혈재관주손상.
    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 (1.5 mL/kg). The method of injection was as same as I/R group.③ Index observation: 4 mL venous blood samples were obtained before ligature, at 40 minutes of ischemia and 40minutes 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.
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