离体心脏长时间在心肌保护液中低温保存对线粒体的影响  

Effects of prolonged cold storage in cardioplegic solution on mitochondria in isolated hearts

作　　者：王一涵 李双磊[2] 何潇一 肖苍松[2] Wang Yihan;Li Shuanglei;He Xiaoyi;Xiao Cangsong(Graduate School of Medical School of Chinese PLA General Hospital,Beijing,100037,China;Department of Cardiovascular Surgery,the Sixth Medical Center,Chinese PLA General Hospital,Beijing 100853,China)

机构地区：[1]解放军医学院研究生院,北京100037 [2]解放军总医院第六医学中心心血管外科,北京100853

出　　处：《中国体外循环杂志》2025年第2期148-152,161,共6页Chinese Journal of Extracorporeal Circulation

基　　金：军委科技委基础加强计划重点研究项目(2019-JCJQ-ZD-195-00)。

摘　　要：目的研究心脏保存液(HTK)对离体大鼠心脏长时间低温保存的效果,观察其在血流动力学和心脏组织结构方面的变化,并初步探讨HTK液低温保存对心肌线粒体的结构和功能的影响。方法24只雄性SD大鼠,体重280~360 g,随机分为3组,分别是假手术组(Sham)、HTK液低温保存8 h组(HTK-8)和12 h组(HTK-12)。利用Langendorff装置记录心脏血流动力学变化、利用苏木精-伊红(HE)染色观察心肌组织的结构改变,通过透射电镜观察心肌细胞线粒体的变化,Western blot免疫印迹方法检测线粒体相关蛋白的表达。结果在血流动力学方面,HTK-8组中心脏泵功能恢复率和最大舒张末压恢复率高于HTK-12组,差异具有统计学意义(P<0.05)。HE染色显示HTK-8组的心肌细胞结构相对较为完整,间质水肿、细胞肿胀和脂肪改变的程度均明显轻于HTK-12组。透射电镜观察心肌线粒体结构,与Sham组相比,HTK-8组和HTK-12组的线粒体呈现肿胀,线粒体嵴紊乱;但是HTK-8组的线粒体改变轻于HTK-12组。线粒体功能方面,随着保存时间的延长,调控线粒体分裂和融合的两种蛋白发生显著变化,提示线粒体功能受损。结论随着离体心脏保存时间的延长,心肌线粒体的结构和功能受损加剧,从而导致心脏整体功能的下降。Objective To investigate the effect of cardioplegic solution(HTK)on isolated rat hearts during prolonged cold storage,observing changes in hemodynamics and cardiac tissue structure,and preliminarily explorring the effects of HTK on myocardial mitochondria under cold storage conditions.Methods 24 male SD rats weighing 280-360 g were randomly divided into three groups:sham group,HTK cold storage for 8 hours(HTK-8)and 12 hours(HTK-12).Cardiac hemodynamic changes were recorded using the Langendorff device,and structural changes in myocardial tissue were observed using HE staining,changes in cardiomyocyte mitochondria were assessed through transmission electron microscopy,and the expression of mitochondria-associated proteins was detected by Western blot.Results In terms of hemodynamics,the recovery rate of pump function and maximum end-diastolic pressure were significantly higher in HTK-8 than in HTK-12,with a statistically significant difference(P<0.05).HE staining showed that the cardiomyocyte structure of HTK-8 was relatively intact,and the degree of interstitial edema,cell swelling and fat changes were significantly less severe than that of HTK-12.Transmission electron microscopy showed mitochondrial swelling and cristae disruption in both HTK-8 and HTK-12 groups,but the changes were less severe in the HTK-8 group.In terms of mitochondrial function,the two proteins regulating mitochondrial fission and fusion changed significantly with the extension of preservation time,suggesting impaired mitochondrial function.Conclusion Prolonged cold storage of isolated hearts exacerbates mitochondrial structural and functional damage,contributing to overall cardiac function.

关 键 词：心脏低温保存 HTK心肌保护液 心肌保护 线粒体 大鼠 

分 类 号：R654.1[医药卫生—外科学]