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

A comparative study of mouse models of systemic inflammatory response after cardiopulmonary resuscitation between asphyxiation and injection of high concentrations of potassium chloride

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 explore the standardized animal models of cardiac arrest cardiopulmonary resuscitation（CPR）in C57BL/6mice.Method：Eighty C57BL/6mice were performed asphyxia method to induce cardiac arrest,65C57BL/6mice were injected high potassium complicating asphyxia to induce cardiac arrest.Both models were randomly divided four groups respectively,groups before CPR,2h,12 hand 24hgroups after ROSC.The rate of spontaneous circulation restoration（ROSC）and survival rate in each group were observed and recorded.The concentration of IL-1β、IL-6、IL-10、IFN-γin bronchoalveolar lavage fluid（BALF）of mice in each group were examined by enzyme-linked immunosorbent assay.The morphological changes and inflammatory reaction of heart,kidneys,and brain tissue were compared in each group.Result：Compared with asphyxia model,the ROSC of high potassium complicating asphyxia model was lower,the survival rate of mice in both 2hand 12 hgroups after ROSC were also lower,but the survival rate in 24 hgroups after ROSC were similar.Compared with the groups before CPR,the concentration of IL-1β、IL-6、IL-10、IFN-γin BALF of 2h,12 h,24hgroups after CPR were increased significantly.Compared with asphyxia model,the inflammatory reaction in 12 hgroup of high potassium complicating asphyxia model was more significantly increased.Compared with groups before CPR,the protein level of phospho-IκB-αin myocardial tissue and kidneys tissue of both two models showed significant increases in 2h、12hand24hgroups after ROSC.Similarly,compared with asphyxia model,the protein level of phospho-IκB-αin kidneys tissue of high potassium complicating asphyxia model were significant increased in 2hand 12 hgroups after ROSC.Conclusion：Asphyxia model in mice is now more commonly used to induce cardiac arrest--cardiopulmonary resuscitation model,because it doesn＇t need any invasive surgery,has a higher rate of ROSC,and a higher survival rate.And the morphological changes of heart,kidneys,brain tissue showed similar changes with high potassium complicating asphyxia model.In the high potassium complicating asphyxia model of our study,the starting time of breath and cardiac arrest can achieve absolute consistency,the damage time of tissue ischemia and hypoxia can be recorded more exactly,and systemic inflammatory response present more significant changes.Therefore,injecting high potassium complicating asphyxia method is deserved to widely use to make standardized cardiac arrest--cardiopulmonary resuscitation model in mice.