亚低温治疗对心肺复苏后鼠脑组织凋亡相关蛋白和胶质纤维酸性蛋白的作用

发布时间：2018-05-17 01:03

本文选题：亚低温 + 心脏骤停　；参考：《苏州大学》2016年博士论文

【摘要】：第一部分:大鼠心肺复苏后亚低温模型的建立与评估目的:利用窒息法使大鼠心脏骤停后行心肺复苏,在复苏后使用冰袋诱导低温,并评估大鼠心肺复苏后的亚低温模型。方法:将10%水合氯醛(0.3 ml/100 g)注射进腹腔内用于麻醉,然后气管切开放置气管插管,将套管插到股动脉和股静脉。然后检测动脉血压以及记录标准Ⅱ号导联动态心电图。在距离肛门1.5厘米处放置探针并使用电子温度计来检测直肠温度。手术后,让大鼠情绪稳定15分钟,然后去除心率和血压明显异常的大鼠。夹住气管,使大鼠突然心脏骤停(心电图表现为心室颤动,无脉性电活动或心搏停止,或收缩压下降到25 mm Hg以下并且动脉搏动消失)。6分钟后,将夹住的气管松开,并且连接动物呼吸机为其辅助通气,同时使用胸外心脏按压,如果有心室纤维性颤动,使用电除颤和利多卡因静脉注射。当自主循环恢复(自主循环标准为HR?170次/分且收缩压?90 mm Hg),并且自主循环维持5分钟后,终止按压。如果在心肺复苏6分钟后,自主循环没有恢复,复苏终止。当自主呼吸恢复时,撤除呼吸机。在监测到自主循环恢复35分钟后,移除股动脉以及静脉插管,缝合。当大鼠苏醒时,立即0.12 ml/100 g浓度为10%的水合氯醛注入腹腔内。在复苏后,对照组在室温下(25℃)治疗,而亚低温两组使用冰袋诱导低温,并且在诱导体温降低后的30分钟内,达到最佳温度(33/30℃)。如果体温偏离最佳温度,予以调节。如果体温高于预期标准,喷稀乙醇或者再结合电风扇使其降低温度。当大鼠的体温低于预期标准,将大鼠放于室温的环境下使其回温。体温的波动幅度要控制在±0.3℃内。结果:大鼠在心肺复苏后的30分钟内,亚低温组分别达到轻度及中度低温温度(33/30℃)。结论:大鼠心肺复苏后轻度及中度低温模型建立成功。第二部分:亚低温治疗对心肺复苏后大鼠脑组织凋亡相关蛋白的作用目的:探讨轻度和中度低温治疗对心肺复苏后大鼠脑组织凋亡相关蛋白的作用。方法:30只SD大鼠被随机分成对照组(正常体温),体温为33℃轻度低温组以及体温为30℃中度低温组,每组各10只。对照组大鼠在进行心肺复苏后,25℃室温下接受常规治疗,而轻度低温组以及中度低温组的大鼠经过心肺复苏后,分别接受33℃和30℃低温治疗。所有组经过心肺复苏24小时后,将脑组织取出进行研究,并利用免疫组化法检测大脑皮层的凋亡相关蛋白caspase-3,用蛋白免疫印迹法(western blot)检测凋亡相关蛋白Bcl-2以及Bax的表达。结果:与对照组相比,大脑神经元亚低温组的caspase-3明显下降(p0.01),体温为30℃的低温组与33℃低温组相比caspase-3明显更低,但无统计学意义(p?0.05);Bcl-2的表达在亚低温组有明显的增加(p0.01),并且体温为30℃低温组的增加程度高于体温为33℃的低温组(p0.05);而Bax的表达在这三组中无明显的区别。结论:轻度及中度低温可以下调caspase-3,上调Bcl-2的表达。显示了低温在心肺复苏之后有保护大脑的作用。并且30℃的低体温疗法比33℃的低温疗法有更好的治疗作用。第三部分:亚低温治疗对心肺复苏后大鼠脑组织胶质纤维酸性蛋白(GFAP)的作用目的:探讨亚低温治疗对心肺复苏后大鼠脑组织胶质纤维酸性蛋白(GFAP)的作用。方法:30只SD大鼠随机分成对照组(正常体温),体温为33℃的低温组以及体温为30℃低温组,每组各10只。对照组的大鼠在进行心肺复苏后,在25℃室温下接受常规治疗,而轻度低温组以及中度低温组的大鼠,经过心肺复苏后在亚低温下进行治疗。在所有组经过心肺复苏24小时后,将脑组织取出研究,并利用免疫组化法检测大脑皮层的胶质纤维酸性蛋白(GFAP)表达。结果:与对照组相比,胶质纤维酸性蛋白的表达在亚低温组中有明显的增加(p0.01)。结论:轻度及中度低温通过减少caspase-3的表达以及增加Bcl-2的表达去调控神经胶质纤维酸性蛋白的表达,引起胶质纤维酸性蛋白的表达上调,从而促进大脑细胞信号转导,进一步抑制细胞凋亡,减少脑损伤。
[Abstract]:The first part: the establishment and evaluation of hypothermia model in rats after cardiopulmonary resuscitation: using asphyxia method to make cardiopulmonary resuscitation after cardiac arrest, using ice bag to induce hypothermia after resuscitation, and evaluate the hypothermia model after cardiopulmonary resuscitation in rats. Method: injection of 10% chloral chloral (0.3 ml/100 g) into the abdominal cavity for anesthesia, and then trachea The cannula was inserted into the endotracheal intubation and inserted into the femoral and femoral veins. Then the arterial blood pressure and the standard II lead dynamic electrocardiogram were detected. A probe was placed at 1.5 cm from the anus and an electronic thermometer was used to detect the rectal temperature. After surgery, the rats were emotionally stabilized for 15 minutes, and then the heart rate and blood pressure were significantly abnormally removed. Rats were trapped in the trachea, causing sudden cardiac arrest in rats (the electrocardiogram showed ventricular fibrillation, no pulse electrical activity or cardiac arrest, or the systolic pressure dropped below 25 mm Hg and the arterial pulsation disappeared). After.6 minutes, the trapped trachea was loosened and the animal ventilator was connected to its auxiliary ventilation, and the chest compression was used, if the heart was pressed, if the heart was in mind. Ventricular fibrillation, using electric defibrillation and lidocaine intravenous injection. When autonomic circulation recovery (independent circulation standard is HR? 170 / sub and systolic pressure? 90 mm Hg), and the autonomic circulation is maintained for 5 minutes, the press is terminated. If the cardiopulmonary resuscitation is 6 minutes, the autonomic circulation is not recovered and the resuscitation terminates. When the self respiration recovery is recovered, the ventilator is removed. After 35 minutes of recovery, the femoral artery and venous cannula were removed and sutured. When the rat was awakened, the 0.12 ml/100 G concentration of chloral hydrate was injected into the abdominal cavity immediately. After resuscitation, the control group was treated at room temperature (25 degrees C), while the hypothermia two groups used ice bags to induce hypothermia and 30 minutes after the hypothermia was induced. Inside, reach the best temperature (33/30 C). If the temperature deviates from the best temperature, adjust it. If the body temperature is higher than the expected standard, spray dilute ethanol or combine the electric fan to reduce the temperature. When the body temperature of the rat is lower than the expected standard, the rat is put back at room temperature. The fluctuation range of body temperature is controlled within 0.3. In 30 minutes after cardiopulmonary resuscitation, mild to moderate hypothermia temperatures (33/30 C) were reached in the hypothermia group. Conclusion: the mild to moderate hypothermia model after cardiopulmonary resuscitation in rats was established successfully. The second part: the effect of mild hypothermia treatment on apoptosis related egg white in the brain tissue of rats after cardiopulmonary resuscitation: the treatment of mild and moderate hypothermia treatment Methods: 30 SD rats were randomly divided into control group (normal body temperature), mild hypothermia at 33 centigrade and 10 moderate hypothermia group at 30 centigrade, 10 rats in each group. The control group received routine treatment at the temperature of 25, and mild hypothermia after cardiopulmonary resuscitation, and the mild hypothermia group. After cardiopulmonary resuscitation, rats in the moderate hypothermia group received 33 and 30 centigrade cryogenic treatment. After 24 hours of cardiopulmonary resuscitation, the brain tissue was taken out and the apoptosis related protein caspase-3 in the cerebral cortex was detected by immunohistochemistry. The apoptosis related protein Bcl-2 was detected by protein immuno trace (Western blot) method. Results: compared with the control group, compared with the control group, the caspase-3 of the hypothermia group of the cerebral neurons decreased significantly (P0.01), the low temperature group at 30 C was significantly lower than the 33 C group, but there was no statistical significance (P? 0.05); the expression of Bcl-2 was significantly increased in the sub hypothermia group (P0.01), and the body temperature was 30 centigrade cryogenic group. The degree is higher than the hypothermia group (P0.05) at 33 degrees centigrade; and the expression of Bax is not distinctly different in these three groups. Conclusion: mild and moderate hypothermia can down regulate the expression of Caspase-3 and increase the expression of Bcl-2. It shows that the hypothermia can protect the brain after cardiopulmonary resuscitation, and the low body temperature therapy at 30 C has better treatment than the cryotherapy at 33. The third part: the effect of mild hypothermia on glial fibrillary acidic protein (GFAP) in the brain tissue of rats after cardiopulmonary resuscitation: To explore the effect of hypothermia therapy on glial fibrillary acidic protein (GFAP) in the brain tissue of rats after cardiopulmonary resuscitation. Methods: 30 SD rats were randomly divided into two groups (normal body temperature), hypothermia group at 33 C and body temperature. After cardiopulmonary resuscitation, the rats in the control group received routine treatment at the room temperature of 25, while the rats in the control group received routine treatment at 25 C, while the mild hypothermia group and the moderate hypothermia group were treated at subhypothermia after cardiopulmonary resuscitation. After 24 hours of cardiopulmonary resuscitation, the brain tissue was removed and immuno histochemical was used in all groups. Results: the expression of glial fibrillary acidic protein (GFAP) in the cerebral cortex was detected. Results: compared with the control group, the expression of glial fibrillary acidic protein was significantly increased in the hypothermia group (P0.01). Conclusion: mild to moderate hypothermia can regulate the expression of glial fibrillary acidic protein by reducing the expression of Caspase-3 and increasing the expression of Bcl-2. Glial fibrillary acidic protein expression is upregulated, thereby promoting signal transduction in brain cells, further inhibiting cell apoptosis and reducing brain damage.
【学位授予单位】：苏州大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：R614

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