窒息大鼠心肺复苏模型的改良及复苏后大鼠脑组织AQP4表达变化的研究

发布时间：2018-01-11 05:29

本文关键词：窒息大鼠心肺复苏模型的改良及复苏后大鼠脑组织AQP4表达变化的研究　出处：《苏州大学》2007年硕士论文　论文类型：学位论文

【摘要】： 第一部分窒息大鼠心肺复苏模型的改良目的:改良窒息大鼠心肺复苏模型;探讨心脏停搏持续时间对自主循环恢复后6h存活率和脑含水量的影响,为后续实验确定合适的心脏停搏时间。方法:健康雄性SD大鼠36只,随机分为4组:对照组(n=6),CA1min组、CA3min组、CA5min组,后三组每组均为10只。对照组仅予气管切开插管、股动、静脉置管,手术后6h处死。其它各组予气管切开插管、股动、静脉置管,呼气末夹闭气管导管模拟窒息,心脏停搏(cardiac arrest , CA)持续1min、3min、5min后实施心肺复苏,应用心电监护仪监测三组大鼠心肺复苏期间平均动脉压(mean arterial pressure,MAP)、心率(heart rate,HR)及体温的动态变化,股动脉采血监测血气的动态变化,并观察三组大鼠自主循环恢复(restoration of spontaneous circulation,ROSC)时间、复苏成功率及ROSC后6小时的存活率及脑含水量(brain water content,BWC)。结果:CA3min组的复苏成功率、ROSC后6h存活率与CA5min组比较,有显著性差异。CA3min组BWC与CA1min比较有显著性差异(P㩳0.05)。结论:1.改进窒息大鼠心肺复苏模型,改良后的模型更加稳定,复苏成功率和ROSC后6h存活率均显著高于未改良前。 2.心脏停搏3min后复苏成功率和ROSC后6h存活率均较高,ROSC后6h的BWC增加明显。利用窒息大鼠进行心肺复苏后脑水肿研究,可选择3min为实验心脏停搏时间。第二部分心肺复苏后大鼠脑组织水通道蛋白-4及其mRNA的动态变化和亚低温对其影响的研究目的:探讨复苏后早期脑水肿的变化规律,AQP4及其mRNA在脑水肿发生、发展中的作用,以及亚低温减轻心肺复苏后脑水肿的可能机制。方法:雄性SD大鼠66只,随机分为3组,分别为对照组、常温复苏组、亚低温组,后两组各自进一步分为5个亚组,即自主循环恢复后0.5、1、3、6和9小时组。建立窒息大鼠心肺复苏模型,应用干湿重法测定自主循环恢复后0.5、1、3、6和9小时脑组织含水量的变化;同时观察各时间点脑组织形态学的变化。利用免疫组织化学染色半定量分析的方法,检测相同观察时点脑组织AQP4表达水平的变化。运用RT-PCR方法检测脑组织AQP4mRNA表达的变化,分析AQP4、AQP4mRNA与BWC的相关性。结果:1.常温组大鼠,ROSC后0.5hBWC开始增加,随着时间的延长BWC明显增加,各观察时点BWC与对照组比较,均有显著性差异(P值0.05)。脑组织形态学的变化与以上结果相符。同时,复苏后脑组织AQP4蛋白水平、AQP4mRNA的表达,也呈现出与BWC变化几乎相同的趋势。相关分析表明,BWC与脑组织AQP4、AQP4mRNA水平表达呈明显正相关,相关系数(r)分别为0.847和0.771,(P0.01)。 2. ROSC后0.5小时,亚低温组大鼠BWC和脑组织AQP4表达水平与常温组同一观察时点比较无明显变化;亚低温作用1h、3h、6h和9h时,上述指标均较相同观察时点常温复苏组明显降低(P值均0.05)。相关分析表明,亚低温作用9h内,BWC与脑组织AQP4、AQP4mRNA表达水平呈明显正相关, r分别为0.769和0.780,( P0.01)。结论:1.心肺复苏后0.5h已出现脑水肿,并随着ROSC时间点的延长,脑水肿逐渐加重。 2.脑组织AQP4及其mRNA在ROSC后0.5h表达上调,并随着ROSC时间点的延长,逐渐增高,脑水肿严重程度和AQP4、AQP4mRNA均呈现明显正相关。提示AQP4mRNA可能通过调节AQP4表达,参与了心肺复苏后脑水肿的过程。 3. ROSC后亚低温干预可减轻脑水肿,其机制可能和亚低温抑制AQP4mRNA,下调AQP4的表达有关。
[Abstract]:The improvement of the model of cardiopulmonary resuscitation in the first part of asphyxiated rats
Objective: to improve the cardiopulmonary resuscitation model of asphyxia rats, and to explore the effect of cardiac arrest duration on 6h survival rate and cerebral water content after the recovery of autonomic circulation, so as to establish suitable cardiac arrest time for subsequent experiments.
Methods: 36 healthy male SD rats were randomly divided into 4 groups: control group (n=6), CA1min group, CA3min group, CA5min group, three groups with 10 rats in each group. The control group only received tracheotomy, femoral artery, venous catheter, surgery after 6h death. Other groups were given trachea incision intubation, femoral artery, venous catheter, end expiratory clamping the endotracheal tube to simulate suffocation, cardiac arrest (cardiac arrest, CA) for 1min, 3min, 5min after CPR, cardiopulmonary resuscitation during the application of ECG monitoring in three groups of rats mean arterial pressure (mean arterial pressure, MAP), heart rate (heart rate, HR) and the dynamic changes of body temperature, the dynamic changes of femoral artery blood gas monitoring, and to observe the three groups of rats (restoration of spontaneous ROSC circulation, ROSC), the survival rate of water and brain 6 hours with the success rate of resuscitation and ROSC (brain water content, BWC).
Results: the successful rate of resuscitation in the CA3min group and the survival rate of 6h after ROSC were compared with that of the CA5min group, and there was a significant difference between BWC and CA1min in.CA3min group (P? 0.05).
Conclusion: 1. the model of cardiopulmonary resuscitation in asphyxiated rats was improved. The modified model was more stable, the success rate of resuscitation and the survival rate of 6h after ROSC were significantly higher than that before the improvement.
2., after cardiac arrest, the success rate of 3min recovery and the survival rate of 6h after ROSC were all higher. The BWC of 6h increased significantly after ROSC. The study of cerebral edema after cardiopulmonary resuscitation can be done by asphyxia rats. 3min can be selected as experimental cardiac arrest time.
The dynamic changes of aquaporin -4 and its mRNA in the brain tissue of rats after cardiopulmonary resuscitation and the effect of mild hypothermia on it in second parts
Objective: To investigate the change rule of brain edema in early stage after resuscitation, the role of AQP4 and mRNA in the development and development of cerebral edema, and the possible mechanism of mild hypothermia to relieve brain edema after cardiopulmonary resuscitation.
Methods: 66 male SD rats were randomly divided into 3 groups, including control group, normothermia group and hypothermia group, the two groups after each further divided into 5 subgroups, namely after the restoration of spontaneous circulation 0.5,1,3,6 and 9 hour group. To establish a rat model of cardiopulmonary resuscitation of asphyxia, changes of spontaneous circulation after 9 hours of recovery of 0.5,1,3,6 and brain water content measured by dry wet weight method; at the same time, the morphological changes of brain tissue were observed at different time points. Using immunohistochemistry and semi quantitative analysis, detection of the same observation point in brain tissue AQP4 expression. Using RT-PCR method to detect the expression of changes in brain tissue AQP4mRNA AQP4 analysis and the correlation between AQP4mRNA and BWC.
Results: 1. normal rats, ROSC 0.5hBWC began to increase, with the extension of time BWC increased significantly, compared with the control at each observation time point of BWC, there were significant differences (P = 0.05). The morphological changes of brain tissue coincided with the results above. At the same time, the recovery of brain tissue AQP4 protein level, the expression of AQP4mRNA also, showing almost the same trend with the change of BWC. Correlation analysis showed that BWC and AQP4 in brain tissue, the level of AQP4mRNA expression was positively correlated, correlation coefficient (R) were 0.847 and 0.771 (P0.01).
0.5 hours after 2. ROSC, hypothermia group BWC and rat brain tissue AQP4 expression level and the normal group at the same observation time there is no obvious change; mild hypothermia effects of 1H, 3h, 6h and 9h, the above indexes were observed in the same normothermia group significantly decreased (P < 0.05). Correlation analysis showed that analysis of the effect of the mild hypothermia in 9h, BWC and AQP4 in brain tissue, the expression level of AQP4mRNA was positively correlated with R, respectively 0.769 and 0.780 (P0.01).
Conclusion: 1. 0.5h has cerebral edema after cardiopulmonary resuscitation, and the brain edema gradually increases with the prolongation of ROSC time.
2., the expression of AQP4 and mRNA in brain tissue increased after ROSC, and increased with the extension of ROSC time. The severity of brain edema was positively correlated with AQP4 and AQP4mRNA. It suggested that AQP4mRNA might participate in the process of cerebral edema after regulating the expression of AQP4 in cardiopulmonary resuscitation.
Hypothermia intervention after 3. ROSC can reduce brain edema, and its mechanism may be related to subhypothermia inhibition of AQP4mRNA and down regulation of the expression of AQP4.

【学位授予单位】：苏州大学
【学位级别】：硕士
【学位授予年份】：2007
【分类号】：R-332

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