七氟醚预处理对老龄大鼠术后认知功能障碍的影响

发布时间：2018-06-04 00:02

本文选题：麻醉学 + 吸入　；参考：《河北医科大学》2014年硕士论文

【摘要】：目的：本研究拟通过探讨卤族类挥发性麻醉剂七氟醚预处理对老龄大鼠术后认知功能障碍的影响情况，为临床相关试验研究及麻醉管理提供参考。方法：健康成年雄性SD大鼠138只，18月龄，体重为400~450g，采用随机数字表法，将其随机分为3组（n=46）：对照组（C组）、手术组(O组)及七氟醚预处理组(Sev组)。 C组：仅对大鼠腹腔注射戊巴比妥钠进行麻醉处理，而不对其进行任何的手术操作。 O组：腹腔注射戊巴比妥钠麻醉大鼠后，对其行剖腹探查手术，即对大鼠进行备皮、消毒，然后经腹部正中切口、用镊子钝性分离腹部肌肉并显露腹膜、解剖剪剪开腹膜并暴露腹腔，应用探查针依次探查大鼠的肝脏、脾脏、胃、小肠及大肠，手术时间共为30min，手术结束后缝合伤口，并于伤口处粘贴透气防水伤口贴膜，最后腹腔注射青霉素，以防止术后伤口发生感染。 Sev组：将大鼠置于有机玻璃麻醉箱内，进气端连接七氟醚挥发罐，以纯氧作为载体气体，出气端连接麻醉气体监测仪，维持麻醉箱内七氟醚的浓度为2.4%，麻醉箱底部铺设少量的钠石灰以用来吸收二氧化碳，持续30min后吸入空气，吸入空气30min后，处理同O组。每组随机选取10只大鼠，分别于术前30min及术后第1、3、5、7天时，行Morris水迷宫实验，并记录大鼠在原平台象限停留的时间、逃避潜伏期和穿越原平台的次数。分别于术前30min及术后第1、7天时，每组随机选取10只大鼠，，麻醉后取海马组织，采用TUNEL细胞凋亡检测技术测定海马神经元的凋亡指数；流式细胞技术测定海马神经元胞浆Ca2+离子浓度（[Ca2+]i）、凋亡率；透射电镜下观察大鼠海马神经元超微结构的病理学改变（线粒体、内质网、高尔基体、胞浆及胞核）。每组随机选取6只大鼠，分别于术前即刻（T0）、术中10min（T1）、20min (T2)、30min（T3）时，记录体温并采集动脉血样行血气分析。结果：3组不同时间点的体温及血气分析各指标均在正常范围内，组间比较差异无统计学意义（P0.05）。与术前30min时比较，O组术后各时点大鼠原平台象限停留时间缩短、逃避潜伏期延长和穿越原平台次数减少，大鼠海马神经元胞浆[Ca2+]i、凋亡指数/凋亡率升高（P0.05）；Sev组术后第1、3、5天时原平台象限停留时间缩短、逃避潜伏期延长和穿越原平台次数减少，术后第1天时大鼠海马神经元胞浆[Ca2+]i、凋亡指数/凋亡率升高（P0.05），Sev组术后第7天时上述各指标差异无统计学意义（P0.05）；C组术后各时点上述各指标差异无统计学意义（P0.05）。与C组比较，O组术后各时点大鼠原平台象限停留时间缩短、逃避潜伏期延长和穿越原平台次数减少，大鼠海马神经元的胞浆的[Ca2+]i、凋亡指数/凋亡率升高（P0.05）；Sev组术后第1、3、5天时原平台象限停留时间缩短、逃避潜伏期延长和穿越原平台次数减少，Sev组术后第1天时大鼠海马神经元胞浆[Ca2+]i、凋亡指数/凋亡率升高（P0.05），Sev组术后第7天时上述各指标差异无统计学意义（P0.05）。与O组比较，Sev组术后各时点大鼠原平台象限停留的时间延长、逃避潜伏期缩短和穿越原平台的次数增加，大鼠海马神经元胞浆[Ca2+]i、凋亡指数/凋亡率降低（P0.05）。电镜下可见C组海马神经元胞浆内各细胞器结构未见明显异常；O组术后第1天时，海马神经元胞质高度水肿，线粒体嵴内外膜融合消失及空泡化，粗面内质网脱颗粒，术后第7天时，胞质水肿减轻，多数线粒体仍表现为空泡化，高尔基体及粗面内质网结构异常；Sev组术后第1天时，电镜下可见海马神经元胞质轻度水肿，线粒体空泡化，粗面内质网结构异常，但并没有扩张及脱颗粒现象，术后第7天时，电子显微镜下可见神经元线粒体空泡化明显减少，粗面内质网结构接近正常。结论：12.4%七氟醚预处理可减轻老龄大鼠术后认知功能障碍。 2其机制与调节钙稳态失衡，降低海马神经元胞浆钙离子浓度，抑制海马神经元的凋亡有关。
[Abstract]:Objective : To study the effect of the pretreatment of halogenated volatile anesthetics on cognitive dysfunction in aged rats , and provide a reference for clinical trial research and anesthesia management .

Methods : 138 , 18 - month - old male SD rats were randomly divided into three groups ( n = 46 ) : control group ( group C ) , operation group ( O group ) and group of heptafluoroether ( Sev group ) .

Group C : The rats were anesthetized by intraperitoneal injection of sodium opental , without any surgical operation .

Group O : After the rats were anesthetized by intraperitoneal injection of sodium opental sodium , the rats were subjected to laparotomy , namely , the rats were prepared and disinfected , then the abdominal muscles were cut through the midline incision of the abdomen , the peritoneum was cut off with forceps , the peritoneum was cut off and the peritoneum was exposed , the operation time was 30 min , the wound was sutured after the operation was completed , and the air - permeable waterproof wound patch was stuck at the wound site , and penicillin was injected intraperitoneally to prevent infection of the wound after operation .

Sev group : the rats were placed in an organic glass anesthesia box , the air inlet end was connected with a heptafluoroether volatilization tank , pure oxygen was used as the carrier gas , the air outlet end was connected with an anesthetic gas monitor , the concentration of the heptafluoroether in the anesthesia box was maintained to be 2.4 % , a small amount of soda lime is laid at the bottom of the anesthesia box to absorb carbon dioxide , and after 30 minutes of inhalation air , the same O group is treated .

Ten rats were randomly selected in each group at 30 min before operation and 1 , 3 , 5 , 7 days after operation respectively , and the time , escape latency and the number of times of crossing the original platform were recorded . 10 rats were randomly selected in each group at 30 min before operation and 1 and 7 days after operation .
Flow cytometry was used to determine the concentration of Ca2 + ion and apoptosis in hippocampal neurons .
The ultrastructural changes of rat hippocampal neurons were observed under transmission electron microscope ( mitochondria , endoplasmic reticulum , golgi , cytoplasm and nucleus ) . Six rats were randomly selected in each group , and the body temperature was recorded and blood gas analysis of arterial blood samples was collected at the time of operation ( T0 ) , 10 min ( T1 ) , 20 min ( T2 ) and 30 min ( T3 ) .

Results : All the indexes of body temperature and blood gas analysis in three groups were within the normal range , and there was no significant difference between the groups ( P0.05 ) .

Compared with the pre - operative 30min , the retention time of the original platform quadrant in the O group was shortened , the escape latency prolonged and the number of crossing the original platform decreased , and the apoptosis index / apoptosis rate of the rat hippocampal neurons increased ( P0.05 ) .
In Sev group , the retention time of the original platform was shortened , the escape latency prolonged and the number of crossing the original platform decreased , and the apoptosis index / apoptosis rate of the rat hippocampal neurons increased ( P0.05 ) at the first day after the operation , and the difference of the above indexes was not statistically significant at the 7th day after the Sev group ( P0.05 ) .
There was no significant difference between the above indexes in group C after operation ( P0.05 ) .

Compared with group C , the retention time of the original platform was shortened , the escape latency prolonged and the number of crossing the original platform decreased , and the apoptosis index / apoptosis rate of rat hippocampal neurons increased ( P0.05 ) .
In Sev group , the retention time of the original platform was shortened , the escape latency prolonged and the number of crossing the original platform decreased , and the apoptosis index / apoptosis rate of the rat hippocampal neurons increased ( P0.05 ) at the first day after the Sev group , and the difference of the above indexes was not statistically significant at the 7th day after the Sev group ( P0.05 ) .

Compared with the O group , the time of stay in the original platform quadrant of the Sev group was prolonged , the escape latency shortened and the number of crossing the original platform increased , and the apoptosis index / apoptosis rate of the rat hippocampal neurons decreased ( P0.05 ) .

Under electron microscope , there was no obvious abnormality in the structure of each organelle in the cytoplasm of the hippocampal neurons in group C .
At the first day after operation , the cytoplasm of the hippocampal neurons was highly edematous , the inner and outer membrane fusion disappeared and vacuolation , the rough endoplasmic reticulum was degranulation . At the 7th day after operation , the cytoplasm edema was relieved . Most of the mitochondria still showed vacuolation , and the golgi and rough endoplasmic reticulum were abnormal .
At day 1 of Sev group , mild edema , vacuolation of mitochondria and abnormal endoplasmic reticulum in hippocampus were observed under electron microscope , but there was no dilatation and degranulation . At 7th day of operation , the vacuolation of mitochondria of neurons decreased obviously , and the structure of rough endoplasmic reticulum was close to normal .

Conclusion : The pretreatment of 12.4 % heptafluoroether can reduce cognitive impairment after aging rats .

The mechanism is related to the regulation of calcium homeostasis , the decrease of calcium ion concentration in the hippocampal neurons , and the inhibition of apoptosis in hippocampal neurons .
【学位授予单位】：河北医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R614

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