含3-硝基-N-甲基水杨酰胺的新型保护液对冷缺血保存离体大鼠心脏的保护  

Cardioprotective effect of 3-nitro-N-methyl salicylamide on the isolated rat heart under cold ischemia preservation

作　　者：王硕 刘文英 吕超凡 李佳聪 耿燚 赵云罡[1] Wang Shuo;Liu Wenying;LüChaofan;Li Jiacong;Geng Yi;Zhao Yungang(Tianjin Key Laboratory of Exercise Physiology and Sports Medicine,Institute of Sports and Health,Tianjin University of Sport,Tianjin 301617,China)

机构地区：[1]天津体育学院运动与健康研究院,天津市运动生理学与运动医学重点实验室,天津市301617

出　　处：《中国组织工程研究》2022年第8期1194-1201,共8页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金项目(31971100),项目负责人:赵云罡。

摘　　要：背景:缺血组织再灌注期间,活性氧过量产生造成的氧化应激损伤是缺血再灌注损伤发生的主要因素。线粒体电子传递链是细胞活性氧的主要来源,因此通过靶向降低线粒体复合体的电子传递速度,将有效减轻因活性氧爆发造成的心肌缺血再灌注损伤。3-硝基-N-甲基水杨酰胺是呼吸链复合体Ⅲ的半抑制剂,可以有效降低电子传递速度,具有心脏保存的潜在应用价值,但目前尚无明确的研究和临床应用。目的:探讨以3-硝基-N-甲基水杨酰胺为主要成分的新型心脏保护液对8 h离体冷缺血保存大鼠心脏的保护效果。方法:取雄性健康Wistar大鼠心脏,稳定灌注30 min后,用不同心脏保护液停跳并低温静置保存8 h,根据所用心脏保存液分为对照组(不保存)、3-NNMS新型保护液组、Celsior保护液组、3-NNMS+Celsior保护液组。各组心脏采用Powerlab生物信号采集系统检测心脏血流动力学指标;提取心肌线粒体检测线粒体力能学指标;采用ELISA法检测心肌损伤标志物心肌肌钙蛋白T、肌酸激酶同工酶MB、乳酸脱氢酶;组织切片法检测心脏形态学变化;采用荧光显微镜检测心肌组织活性氧水平。结果与结论:①3-硝基-N-甲基水杨酰胺可显著促进心率恢复水平(P<0.05),并减少外周灌流液上清中心肌损伤相关蛋白心肌肌钙蛋白T、肌酸激酶同工酶MB表达水平(P<0.05);②3-硝基-N-甲基水杨酰胺可显著提高线粒体膜电位水平(P<0.05),有效维持线粒膜结构;③3-硝基-N-甲基水杨酰胺可显著提高超氧化物歧化酶活性(P<0.05),增强心肌抗氧化功能,降低氧化损伤;④提高3-硝基-N-甲基水杨酰胺使用浓度,对心肌细胞培养不会产生毒性作用,表明其不存在使用剂量安全性问题;⑤结果表明,3-硝基-N-甲基水杨酰胺可通过提高抗氧化酶活性、促进心肌活性氧清除、达到降低氧化应激及其诱导的心肌损伤的效果;同时3-硝基-N-甲基水杨BACKGROUND: During reperfusion, increased reactive oxygen species will cause oxidative stress injury in ischemic organs, which is the critical factor for ischemia and reperfusion injury. Cellular reactive oxygen species are mainly originated from mitochondrial electron transport chain. Therefore, it is an effective strategy to reduce myocardial ischemia/reperfusion injury due to elevated reactive oxygen species by inhibiting electron transport at mitochondrial electron transport chain. 3-Nitro-N-methyl-salicylamide(3-NNMS) is a semi-inhibitor for electron transport chain complex Ⅲ, which can slow down the electron transport rate. 3-NNMS has potential applications for cardiac preservation;nevertheless, no related definite research or clinical practice has been reported.OBJECTIVE: To investigate the protective effect of a new cardioplegic solution with 3-NNMS as the main component on isolated rat heart preservation for 8 hours. METHODS: Heart specimens were taken from healthy male Wistar rats, perfused stably for 30 minutes, and then preserved in different cardioplegic solutions at low temperature for 8 hours. According to different cardioplegic solutions used, rat heart specimens were divided into a control group(no preservation), a 3-NNMS cardioplegic solution group, a Celsior cardioplegic group, and a 3-NNMS+Celsior cardioplegic solution group. The hemodynamic changes of the heart were detected by Powerlab instruments. The mitochondrial function in the preserved myocardium was measured by Oxygraph-2 k High-resolution respirometry and chemiluminescence apparatus. Myocardial injury was assessed by detecting the expression of cardiac troponin T, creatine kinase isoenzyme MB, and lactate dehydrogenase using ELISA. The morphological changes of the heart were histologically observed. And the myocardial reactive oxygen species level was tested by inverted fluorescence microscope. RESULTS AND CONCLUSION: The 3-NNMS cardioplegic solution could improve the heart rate recovery compared with the Celsior(P < 0.05), and dec

关 键 词：3-硝基-N-甲基水杨酰胺 心肌缺血再灌注损伤 线粒体抑制剂 心脏保护液 离体心脏保存 活性氧 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学]