窒息法与高钾合并窒息法制备心肺复苏后全身炎症反应小鼠模型的比较研究

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目的:探讨C57BL/6小鼠心搏骤停心肺复苏标准化动物模型,为研究心搏骤停后全身炎症反应综合征(SIRS)、多器官功能障碍综合征(MODS)建立实验室基础和必要条件。方法:随机选取80只C57BL/6小鼠应用窒息法制备心肺复苏模型(窒息组),65只C57BL/6小鼠应用高钾合并窒息法制备心肺复苏模型(高钾合并窒息组),将2组小鼠再随机分为复苏前组、复苏后2、12、24h组。观察各组小鼠自主循环恢复(ROSC)率和生存率;运用酶联免疫吸附法(ELISA)检测各组肺泡灌洗液IL-1β、IL-6、IL-10、IFN-γ浓度;比较复苏后心脏、肾脏、脑组织形态学改变以及全身炎症反应程度。结果:与窒息组比较,高钾合并窒息组ROSC率较低,复苏后2、12h小鼠存活率较低,复苏后24h小鼠存活率一致。与复苏前比较,2组模型在复苏后2、12、24h肺泡灌洗液中IL-1β、IL-6、IL-10、IFN-γ的浓度均显著上升。与窒息组比较,高钾合并窒息组在复苏后12h炎症反应表现更突出,2组在复苏后24h呈现下降趋势,但仍显著高于心肺复苏前的基线水平。与复苏前比较,2组模型在复苏后2、12、24h心肌组织和肾脏组织磷酸化IκB-α(p-IκB-α)蛋白表达显著上升,与窒息组复苏后同一时间点比较,高钾合并窒息组在复苏后2、12h肾脏组织p-IκB-α蛋白显著较高。结论:窒息模型是现今制备小鼠心肺复苏模型中较为常用的方法,不需要任何有创的外科手术,模型复苏成功率高,心脏、肾脏、脑组织形态学改变与高钾合并窒息组一致。高钾合并窒息小鼠心肺复苏模型,呼吸心跳骤停的起始时间可以达到完全一致,组织缺血缺氧损伤的时间更确切,全身炎症反应更显著,是值得推广应用的小鼠心搏骤停心肺复苏标准化动物模型。 Objective: To investigate the standard animal model of cardiopulmonary resuscitation (CPR) in C57BL / 6 mice. To establish a laboratory basis and necessary conditions for the study of systemic inflammatory response syndrome (SIRS) and multiple organ dysfunction syndrome (MODS) after cardiac arrest . Methods: A total of 80 C57BL / 6 mice were randomly selected to establish a cardiopulmonary resuscitation model (asphyxia group) with asphyxia. Sixty-five C57BL / 6 mice were subjected to cardiopulmonary resuscitation (hyperkalemic asphyxia group) Groups of mice were randomly divided into pre-recovery group, 2,12,24 h after resuscitation group. The rate of spontaneous circulation recovery (ROSC) and the survival rate of mice in each group were observed. The concentrations of IL-1β, IL-6, IL-10 and IFN-γ in BALF were detected by enzyme-linked immunosorbent assay After resuscitation heart, kidney, brain morphological changes and systemic inflammatory response. Results: Compared with the asphyxia group, the ROSC rate in the high potassium combined asphyxia group was lower than that in the asphyxia group, the survival rate of the mice was lower at 2 and 12 hours after resuscitation, and the survival rate was the same at 24h after resuscitation. Compared with those before resuscitation, the concentrations of IL-1β, IL-6, IL-10 and IFN-γ in BALF in 2, 12 and 24 h after resuscitation in both groups were significantly increased. Compared with the asphyxia group, the inflammatory response of the hyperkalemic asphyxia group was more prominent at 12h after resuscitation, while the two groups showed a decreasing trend 24h after resuscitation, but still significantly higher than the baseline before cardiopulmonary resuscitation. Compared with that before resuscitation, the expression of phosphorylated IκB-α (p-IκB-α) in myocardium and kidney increased significantly at 2, 12 and 24 hours after resuscitation in both groups. Compared with the same time after resuscitation, In the asphyxia group, the p-IκB-α protein in kidney tissue was significantly higher at 2 and 12 hours after resuscitation. Conclusion: The asphyxial model is the most commonly used method for preparing CPR model in mice. It does not require any invasive surgery. The success rate of model resuscitation is high. The morphological changes of heart, kidney and brain are consistent with those of hyperkalemic asphyxia group. High potassium asphyxia mouse CPR model, respiratory arrest start time can be completely consistent, tissue ischemia and hypoxia injury more precise time, more systemic inflammatory response is worth promoting the use of mouse heart steps Stop CPR standardized animal model.
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