乳大鼠心肌细胞糖氧剥夺-营养恢复模型模拟在体缺血/再灌注损伤的可行性研究  

Feasibility Analysis of Oxygen Glucose Deprivation-nutrition Resumption on Neonatal Rat Cardiomyocytes in Vitro Model of Myocardial Ischemia-Reperfusion Injury

作　　者：杨桂珍 薛富善 刘亚洋 李慧娴 刘庆 廖旭 YANG Gui-zhen;XUE Fu-shan;LIU Ya-yang;LI Hui-xian;LIU Qing;LIAO Xu(Department of Anesthesiology,Plastic Surgery Hospital,Chinese Academy of Medical Sciences and Peking Union Medical College,Beijing 100144,China)

机构地区：[1]中国医学科学院整形外科医院麻醉科,北京市100144

出　　处：《中国分子心脏病学杂志》2019年第1期2764-2769,共6页Molecular Cardiology of China

基　　金：中国医学科学院整形外科医院院所重大基金项目(Z2017001)

摘　　要：目的乳大鼠心肌细胞糖氧剥夺-营养恢复(OGD-NR)模型是最常用的模拟心肌缺血-再灌注损伤(myocardial ischemiareperfusion injury,MIRI)的离体建模方法之一。但是,至今尚无研究对该模型的可行性进行评估。该实验旨在评价乳大鼠心肌细胞OGD-NR离体心肌细胞模型模拟在体MIRI的可行性。方法将乳大鼠心肌细胞随机分成对照(C)组、模拟缺血(SI)组和模拟缺血再灌注(SIR)组。实验结束时检测细胞形态学、乳酸脱氢酶(LDH)释放情况、三磷酸腺苷(ATP)水平、活性氧(ROS)水平、线粒体膜电位(MMP)水平以及炎症因子水平,以评价各组细胞损伤的表型和特点。结果与C组相比,SI组的心肌细胞形态学出现损伤、LDH释放显著增加(P<0.05)、ATP明显降低(P<0.05)、ROS生成增加(P<0.05)、MMP水平降低(P<0.05)。与SI组相比,SIR组的心肌细胞形态学并未进一步恶化,LDH释放明显降低(P<0.05)、ATP水平明显增高(P<0.05)。与SI组相比,相应的SIR组未发生大量ROS生成、MMP坍塌以及过度的炎症反应。结论乳大鼠心肌细胞OGD-NR不能成功模拟在体MIRI的特征,即对发生SI损伤的心肌细胞实施SIR处理后,形态学上未出现进一步的损伤、LDH释放明显降低、ATP明显增高、无大量ROS生成、无MMP坍塌且没有过度炎症反应发生。Objective Oxygen glucose deprivation nutrition resumption (OGD-NR) model on neonatal rat cardiomyocytes (NRCs) is commonly used in vitro model of simulated myocardial ischemia-reperfusion injury (MIRI), but there has been no study to assess whether this model can effectively imitate the characteristics of MIRI in vivo. This experiment was designed to assess the feasibility of this model simulating MIRI. Methods NRCs were randomly divided into control (C), simulated ischemia (SI) and simulated ischemia-reperfusion (SIR) groups, and cell morphology, lactate dehydrogenase (LDH) release, adenosine triphosphate (ATP) levels, reactive oxygen species (ROS), mitochondrial membrane potential (MMP) and inflammatory cytokines were examined to evaluate the phenotype and characteristics of cell injury. Results Compared with the C group, the cells suffered from morphological damage, LDH release significantly increased (P<0.05), ATP evidently decreased (P<0.05), ROS production significantly increased (P<0.05) and MMP levels obviously decreased in the SI group (P<0.05). Compared with the SI group, however, cellular morphology did not worsen, LDH release significantly decreased (P<0.05), and ATP level evidently increased (P<0.05) in the SIR group. In addition, large generation of ROS, MMP collapse, and over-inflammatory response did not occur in the SIR group compared with the SI group. Conclusion OGD-NR of neonatal rat cardiomyocytes could not successfully simulate the characteristics of MIRI in vivo, as demonstrated by no further damage in morphology, significant decrease in LDH release, significant increase in ATP, no large generation of ROS, no MMP collapse, and no over-inflammatory response by SIR treatment after SI treatment.

关 键 词：心肌缺血-再灌注损伤 离体细胞模型 可行性 

分 类 号：R54[医药卫生—心血管疾病]