本文選題：麻醉學(xué) + 吸入��； 參考：《河北醫(yī)科大學(xué)》2014年碩士論文

【摘要】：目的：本研究擬通過探討鹵族類揮發(fā)性麻醉劑七氟醚預(yù)處理對老齡大鼠術(shù)后認知功能障礙的影響情況，為臨床相關(guān)試驗研究及麻醉管理提供參考。 方法：健康成年雄性SD大鼠138只，18月齡，體重為400~450g，采用隨機數(shù)字表法，將其隨機分為3組（n=46）：對照組（C組）、手術(shù)組(O組)及七氟醚預(yù)處理組(Sev組)。 C組：僅對大鼠腹腔注射戊巴比妥鈉進行麻醉處理，而不對其進行任何的手術(shù)操作。 O組：腹腔注射戊巴比妥鈉麻醉大鼠后，對其行剖腹探查手術(shù)，即對大鼠進行備皮、消毒，然后經(jīng)腹部正中切口、用鑷子鈍性分離腹部肌肉并顯露腹膜、解剖剪剪開腹膜并暴露腹腔，應(yīng)用探查針依次探查大鼠的肝臟、脾臟、胃、小腸及大腸，手術(shù)時間共為30min，手術(shù)結(jié)束后縫合傷口，并于傷口處粘貼透氣防水傷口貼膜，最后腹腔注射青霉素，以防止術(shù)后傷口發(fā)生感染。 Sev組：將大鼠置于有機玻璃麻醉箱內(nèi)，進氣端連接七氟醚揮發(fā)罐，以純氧作為載體氣體，出氣端連接麻醉氣體監(jiān)測儀，維持麻醉箱內(nèi)七氟醚的濃度為2.4%，麻醉箱底部鋪設(shè)少量的鈉石灰以用來吸收二氧化碳，持續(xù)30min后吸入空氣，吸入空氣30min后，處理同O組。 每組隨機選取10只大鼠，分別于術(shù)前30min及術(shù)后第1、3、5、7天時，行Morris水迷宮實驗，并記錄大鼠在原平臺象限停留的時間、逃避潛伏期和穿越原平臺的次數(shù)。分別于術(shù)前30min及術(shù)后第1、7天時，每組隨機選取10只大鼠，，麻醉后取海馬組織，采用TUNEL細胞凋亡檢測技術(shù)測定海馬神經(jīng)元的凋亡指數(shù)；流式細胞技術(shù)測定海馬神經(jīng)元胞漿Ca2+離子濃度（[Ca2+]i）、凋亡率；透射電鏡下觀察大鼠海馬神經(jīng)元超微結(jié)構(gòu)的病理學(xué)改變（線粒體、內(nèi)質(zhì)網(wǎng)、高爾基體、胞漿及胞核）。每組隨機選取6只大鼠，分別于術(shù)前即刻（T0）、術(shù)中10min（T1）、20min (T2)、30min（T3）時，記錄體溫并采集動脈血樣行血氣分析。 結(jié)果：3組不同時間點的體溫及血氣分析各指標(biāo)均在正常范圍內(nèi)，組間比較差異無統(tǒng)計學(xué)意義（P0.05）。 與術(shù)前30min時比較，O組術(shù)后各時點大鼠原平臺象限停留時間縮短、逃避潛伏期延長和穿越原平臺次數(shù)減少，大鼠海馬神經(jīng)元胞漿[Ca2+]i、凋亡指數(shù)/凋亡率升高（P0.05）；Sev組術(shù)后第1、3、5天時原平臺象限停留時間縮短、逃避潛伏期延長和穿越原平臺次數(shù)減少，術(shù)后第1天時大鼠海馬神經(jīng)元胞漿[Ca2+]i、凋亡指數(shù)/凋亡率升高（P0.05），Sev組術(shù)后第7天時上述各指標(biāo)差異無統(tǒng)計學(xué)意義（P0.05）；C組術(shù)后各時點上述各指標(biāo)差異無統(tǒng)計學(xué)意義（P0.05）。 與C組比較，O組術(shù)后各時點大鼠原平臺象限停留時間縮短、逃避潛伏期延長和穿越原平臺次數(shù)減少，大鼠海馬神經(jīng)元的胞漿的[Ca2+]i、凋亡指數(shù)/凋亡率升高（P0.05）；Sev組術(shù)后第1、3、5天時原平臺象限停留時間縮短、逃避潛伏期延長和穿越原平臺次數(shù)減少，Sev組術(shù)后第1天時大鼠海馬神經(jīng)元胞漿[Ca2+]i、凋亡指數(shù)/凋亡率升高（P0.05），Sev組術(shù)后第7天時上述各指標(biāo)差異無統(tǒng)計學(xué)意義（P0.05）。 與O組比較，Sev組術(shù)后各時點大鼠原平臺象限停留的時間延長、逃避潛伏期縮短和穿越原平臺的次數(shù)增加，大鼠海馬神經(jīng)元胞漿[Ca2+]i、凋亡指數(shù)/凋亡率降低（P0.05）。 電鏡下可見C組海馬神經(jīng)元胞漿內(nèi)各細胞器結(jié)構(gòu)未見明顯異常；O組術(shù)后第1天時，海馬神經(jīng)元胞質(zhì)高度水腫，線粒體嵴內(nèi)外膜融合消失及空泡化，粗面內(nèi)質(zhì)網(wǎng)脫顆粒，術(shù)后第7天時，胞質(zhì)水腫減輕，多數(shù)線粒體仍表現(xiàn)為空泡化，高爾基體及粗面內(nèi)質(zhì)網(wǎng)結(jié)構(gòu)異常；Sev組術(shù)后第1天時，電鏡下可見海馬神經(jīng)元胞質(zhì)輕度水腫，線粒體空泡化，粗面內(nèi)質(zhì)網(wǎng)結(jié)構(gòu)異常，但并沒有擴張及脫顆�，F(xiàn)象，術(shù)后第7天時，電子顯微鏡下可見神經(jīng)元線粒體空泡化明顯減少，粗面內(nèi)質(zhì)網(wǎng)結(jié)構(gòu)接近正常。 結(jié)論：12.4%七氟醚預(yù)處理可減輕老齡大鼠術(shù)后認知功能障礙。 2其機制與調(diào)節(jié)鈣穩(wěn)態(tài)失衡，降低海馬神經(jīng)元胞漿鈣離子濃度，抑制海馬神經(jīng)元的凋亡有關(guān)。
[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 .
【學(xué)位授予單位】：河北醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R614

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