水芹乙酸乙酯提取物保护大鼠缺血再灌注损伤心肌的作用

Protective effect of ethyl acetate extract from oenanthe aquatic on injured myocardia in rats after ischemic reperfusion

目的:观察水芹乙酸乙酯提取物对缺血再灌注大鼠心肌组织中谷胱甘肽过氧化物酶的活力、心肌线粒体Ca2+的含量、心肌细胞膜Na+-K+-ATP酶和Mg2+-ATP酶活力的影响,探讨该提取物对缺血再灌注损伤心肌的保护作用。方法:①实验于2000-04/2001-04在延边大学基础医学院机能学实验中心完成。选用健康Wistar大鼠40只。将大鼠随机分为4组:假手术组、模型组、水芹乙酸乙酯提取物组、阳性药物组,每组10只。水芹乙酸乙酯提取物(由水芹提取而得)组、模型组、阳性药物组均结扎冠状动脉左前降支,造成心肌缺血40min再灌注损伤30min模型。药物组舌下静脉给予水芹乙酸乙酯提取物(20mg/kg),模型组舌下静脉给予生理盐水2mL,阳性药物组舌下静脉给予维拉帕米(0.2mg/kg)。假手术组只开胸,分离冠状动脉,不予结扎,不给药。其他各组均于造模前5min给药。②采用还原型谷胱甘肽二硫双硝基苯甲酸定量测定法及其法则测定和计算各组样品中谷胱甘肽过氧化物酶的活力。在原子吸收分光光度计上以火焰法测定心肌线粒体内Ca2+的含量。按Jones法测定和计算心肌细胞膜Na+-K+-ATP酶和Mg2+-ATP酶的活力。③组间计量资料比较采用t检验。结果:大鼠40只均进入结果分析。①心肌组织中谷胱甘肽过氧化物酶活力:模型组明显低于假手术组(P<0.01);而水芹乙酸乙酯提取物组和阳性药物组均明显高于模型组(P<0.01,0.05)。②心肌线粒体内Ca2+含量:模型组明显高于假手术组(P<0.01),而水芹乙酸乙酯提取物组和阳性药物组均明显低于模型组(P<0.01,0.05)。③心肌细胞膜Na+-K+-ATP酶的活力:模型组明显低于假手术组(P<0.01),水芹乙酸乙酯提取物组明显高于模型组(P<0.05)。④心肌细胞膜Mg2+-ATP酶的活力:模型组明显低于假手术组(P<0.01),水芹乙酸乙酯提取物组和阳性药物组均明显高于模型组(P<0.01,0.05)。结论:水芹乙酸乙酯提取物能够增加心肌组织中谷胱甘肽过氧化物酶的活力,降低心肌线粒体Ca2+的含量,增加心肌细胞膜Na+-K+-ATP酶和Mg2+-ATP酶的活力,是保护心肌缺血再灌注损伤心肌的主要机制之一。

AIM： To study the protective effect of ethyl acetate extract from oenanthe aquatic on myocardial ischemia reperfusion injury and its effects on the activity of glutathione peroxidase （GSH-Px）, the content of calcium in myocardial mitochondrials, and the activity of Sosium-Potassium ATPase （Na^＋-K^＋-ATPase） and magnesium-ATPase in cellular membrane of myocardium. METHODS： （1）The experiment was preformed in the Lab Center of Function, Basic Medical College, Yanbian University from April 2000 to April 2001. Forty healthy Wistar rats were selected and divided randomly into 4 groups： sham-operated group, model group, oenanthe aquatic group and positive medicine group with 10 rats in each group. The coronary artery left circumflex of ethyl acetate extract were ligated from oenanthe aquatic group, model group and positive medicine group to make myocardial ischemia for 40 minutes with reperfusion for 30 minutes models. Ethyl acetate extract from oenanthe aquatic was administered in ranine vein of rats in the medicine group （20 mg/kg）. 2 mL saline was treated to the rats in the model group. Verapamil was given to rats in the positive medicine group （0.2 mg/kg）. The rats were only opened the chest and separate the coronary artery in the sham-operated group without ligation and medicine. Those in the other groups were administered with medicine before making models ahead of 5 minutes. （2）The activity of GSH-Px was calculated with GSH-dithiowo nitro-benxoic acid assay method and its rule. The content of Ca^2＋ in myocardium was detected with flame in atom light photometer. The activity of Na^＋-K^＋-ATPase and Mg^2＋-ATPase was calculated by Jones way. （3）T-test was adopted in group measurement data. RESULTS： Forty rats were all involved in the result analysis. （1）The activity of GSH-Px：That in the model group was Obviously lower than that in the sham-operated group （P〈0.01）; But that in the ethyl acetate extract from oenanthe aquatic group and positive medicine group was higher obviously than that in the model group （P〈0.05,0.01）. （2）The content of calcium in myocardial mitochondrials： That in the model group was higher markedly than that in the sham-operated group （P〈0.01）, hut that in the ethyl acetate extract from oenanthe aquatic group and positive medicine group was lower remarkably than that in the model group （P〈0.05,0.01）. （3）The activity of Na^＋-K^＋-ATPase： That in the model group was lower distinctly than that in the sham-operated group （P 〈 0.01）, but that in the ethyl acetate extract from oenanthe aquatic group was higher clearly than that in the model group （P 〈 0.05）.（4） The activity of Mg^2＋-ATPase： That in the model group was lower significantly than that in the sham-operated group （P 〈 0.01）, but that in the ethyl acetate extract from oenanthe aquatic group and positive medicine group was higher obviously than that in the model group （P〈0.01,0.05）. CONCLUSION： Ethyl acetate extract from oenanthe aquatic can increase the activity of GSH-Px of myocardium, reduce the content of Ca^2＋ of my oeardial mitoehondrials and increase the activity of Na^＋-K^-ATFase and Mg^＋-ATPase of cellular membranes. It is the one of mechanisms of protecting the myocardial isehemieal reperfusion injury myocardium.