本文選題：異氟醚 + 發(fā)育期大腦　； 參考：《復(fù)旦大學(xué)》2014年碩士論文

【摘要】：第一部分異氟醚麻醉損傷新生小鼠海馬并引起學(xué)習(xí)記憶障礙目的：探討異氟醚對新生小鼠海馬的損傷及學(xué)習(xí)記憶能力的影響。方法：健康新生7日齡C57BL/6小鼠,隨機(jī)分為兩組：異氟醚組和對照組。異氟醚組：出生第7日起開始進(jìn)行濃度為1.5%的異氟醚麻醉,麻醉三天(出生第7、8、9日),每天2小時(shí),吸入氧濃度100%；對照組在相應(yīng)的時(shí)間僅吸入100%氧氣,麻醉過程中保溫保濕。兩組在第9日麻醉后立即處死,解剖取腦,分離海馬,行Western Blot檢測海馬組織凋亡標(biāo)志蛋白Caspase-3; TUNEL染色計(jì)數(shù)海馬區(qū)凋亡細(xì)胞；Morris水迷宮實(shí)驗(yàn)檢測以后的學(xué)習(xí)記憶能力。結(jié)果：與對照組相比,異氟醚組小鼠海馬組織Caspase-3表達(dá)顯著增加,TUNEL染色海馬區(qū)陽性細(xì)胞數(shù)明顯增多,差異均有統(tǒng)計(jì)學(xué)意義(P0.05)；水迷宮實(shí)驗(yàn)結(jié)果,與對照組相比,異氟醚組小鼠逃避潛伏期明顯延長,平臺(tái)穿越次數(shù)顯著減少,差異有統(tǒng)計(jì)學(xué)意義(P0.05)。結(jié)論：異氟醚麻醉新生小鼠可致其海馬區(qū)神經(jīng)細(xì)胞凋亡,引起海馬損傷,影響大腦發(fā)育,進(jìn)而引起其以后的學(xué)習(xí)記憶能力減退。第二部分異氟醚通過激活FAS信號途徑損傷新生小鼠海馬并引起學(xué)習(xí)記憶障礙目的：探討異氟醚可通過激活FAS信號途徑損傷新生小鼠海馬損傷并引起學(xué)習(xí)記憶能力障礙。方法：健康新生7日齡C57BL/6小鼠,隨機(jī)分為兩組,分別為異氟醚組和空白對照組。兩組麻醉方法同第一部分。在三天麻醉后,脫頸處死,取海馬,行Western Blot檢測FAS蛋白(CD95)和FAS配體蛋白(FASligand,FASL)。另取FAS或FASL基因敲除的B6. MRL-Faslpr/Jnju小鼠和B6Smn. C3-Faslgld/Jnju小鼠作為FAS基因缺陷組和FASL基因缺陷組,對兩種小鼠分別做吸入異氟醚和單純氧氣的處理,用第一部分中相同方法麻醉和單純吸氧三天。用Western Blot方法檢測四組小鼠(FAS基因敲除異氟醚組、FAS基因敲除純氧組、FASL基因敲除異氟醚組和FASL基因敲除純氧組)的Caspase-3的表達(dá)。用TUNEL染色法觀察FAS基因敲除異氟醚組和FASL基因敲除異氟醚組小鼠海馬區(qū)陽性細(xì)胞數(shù)。用水迷宮實(shí)驗(yàn)檢測FAS基因敲除異氟醚組和FASL基因敲除異氟醚組小鼠的學(xué)習(xí)記憶能力。將三個(gè)實(shí)驗(yàn)的結(jié)果與第一部分中相應(yīng)健康異氟醚組和對照組的結(jié)果合并、對比觀察并統(tǒng)計(jì)分析。結(jié)果：異氟醚組麻醉的健康新生小鼠其FAS蛋白和FASL蛋白均較空白對照組表達(dá)顯著增加,差異有統(tǒng)計(jì)學(xué)意義(p0.05)。異氟醚健康組Caspase-3表達(dá)較FAS缺陷異氟醚組和FASL缺陷異氟醚組顯著增加,差異有統(tǒng)計(jì)學(xué)意義(p0.05)；而對照組分別與FAS缺陷純氧組、FAS缺陷異氟醚組、FASL缺陷純氧組和FASL缺陷異氟醚組相比,Caspase-3表達(dá)差異均無統(tǒng)計(jì)學(xué)意義；Two-way ANOVA分析可得異氟醚麻醉與FAS或FASL基因敲除在Caspase-3的表達(dá)影響上有聯(lián)系,FAS或FASL基因敲除可能減弱了異氟醚所引起的Caspase-3表達(dá)的增加。異氟醚健康組TUNEL染色陽性細(xì)胞數(shù)較FAS缺陷異氟醚組和FASL缺陷異氟醚組均顯著增加；而對照組與FAS缺陷異氟醚組和FASL缺陷異氟醚組的TUNEL陽性細(xì)胞數(shù)相比,差異無統(tǒng)計(jì)學(xué)意義。異氟醚健康組的逃避潛伏期較FAS缺陷異氟醚組和FASL缺陷異氟醚組明顯延長,平臺(tái)穿越次數(shù)明顯減少,差異有統(tǒng)計(jì)學(xué)意義(p0.05)；而對照組與該兩組小鼠的差異均無統(tǒng)計(jì)學(xué)意義。結(jié)論：異氟醚可通過激活FASL-FAS信號途徑引起健康新生小鼠海馬區(qū)細(xì)胞的凋亡,從而損傷海馬并引起其以后的學(xué)習(xí)記憶能力下降。對于切斷FASL-FAS信號通路的小鼠則可減輕該種損傷,學(xué)習(xí)記憶能力也有明顯改善。從而證明FAS通路是異氟醚損傷新生小鼠海馬的機(jī)制之一。
[Abstract]:Part 1: the effects of isoflurane on the hippocampus and learning and memory impairment in newborn mice: the effects of isoflurane on hippocampal damage and learning and memory ability of newborn mice. Methods: 7 days old C57BL/6 mice were randomly divided into two groups: isoflurane group and control group: isoflurane group: the concentration of isoflurane began to be concentrated on the seventh day of birth. 1.5% isoflurane anesthesia, three days of anesthesia (7,8,9 day of birth), 2 hours a day, inhaled oxygen concentration 100%; the control group was only inhaled 100% oxygen at the corresponding time and kept warm and moisturizing during the anesthesia. The two groups died immediately after ninth days of anesthesia, dissected the brain, separated the hippocampus, and Western Blot was used to detect the apoptosis marker protein Caspase-3 of the hippocampus; T UNEL staining was used to count the apoptotic cells in the hippocampus and the learning and memory ability after the Morris water maze test. Results: compared with the control group, the expression of Caspase-3 in the hippocampus of the isoflurane group increased significantly, and the number of positive cells in the hippocampus of the TUNEL stained hippocampus increased significantly (P0.05). The results of water maze experiment and control were compared with the control group. Compared with the group, the escape latency of the isoflurane group was significantly prolonged, and the number of platform crossing times decreased significantly (P0.05). Conclusion: Isoflurane can induce neuronal apoptosis in the hippocampus, damage the hippocampus, affect the brain development, and induce the impairment of learning and memory. Second parts of the isoflurin can be induced by isoflurane. Ether can damage the hippocampus of newborn mice by activating FAS signal pathway and cause learning and memory impairment: Isoflurane can damage hippocampus damage in newborn mice by activating FAS signal pathway and cause learning and memory impairment. Methods: healthy newborn 7 day old C57BL/6 mice were randomly divided into two groups, isoflurane group and blank control group, respectively. The two groups were anesthetized with the first part. After three days of anesthesia, they were removed from the neck and took the hippocampus, and the FAS protein (CD95) and the FAS ligand protein (FASligand, FASL) were detected by Western Blot. The B6. MRL-Faslpr/Jnju mice of FAS or FASL knockout were taken as the gene defect group and the gene defect group, and two mice were used. The expression of Caspase-3 in four groups of mice (FAS knockout isoflurane group, FAS knockout pure oxygen group, FASL gene knockout isoflurane group and FASL gene knockout pure oxygen group) was detected by Western Blot method, with the same method of inhaling isoflurane and simple oxygen, respectively. The expression of Caspase-3 in the FAS gene knockout isoflurane group, the FAS knockout isoflurane group, the FASL gene knockout isoflurane group and the FASL gene knockout group of pure oxygen group. The number of positive cells in the hippocampus of FAS gene knockout isoflurane group and FASL gene knockout isoflurane group. The learning and memory ability of FAS gene knockout isoflurane group and FASL gene knockout isoflurane group was detected by water maze test. The results of the three experiments were combined with the results of the Xiang Yingjian Kang isoflurane group and the control group in the first part, and the results were combined with the results of the first part. Results: the expression of FAS protein and FASL protein in the healthy newborn mice anaesthetized by isoflurane increased significantly than that in the blank control group. The difference was statistically significant (P0.05). The expression of Caspase-3 in isoflurane healthy group was significantly higher than that of FAS deficient isoflurane group and FASL deficient isoflurane group (P0.05). Compared with the FAS defect pure oxygen group, the FAS deficient isoflurane group, the FASL defect pure oxygen group and the FASL deficient isoflurane group, there was no significant difference in the Caspase-3 expression, and the Two-way ANOVA analysis could be linked to the expression of FAS or FASL gene knockout in Caspase-3, and the FAS or FASL gene knockout might be weakened. The expression of Caspase-3 induced by isoflurane increased. The number of positive TUNEL staining cells in isoflurane group was significantly higher than that in the FAS deficient isoflurane group and the FASL deficient isoflurane group, while the control group had no statistical difference compared with the number of TUNEL positive cells in the FAS deficient isoflurane group and the FASL deficient isoflurane group. The escape latency was significantly longer than that of FAS defect isoflurane group and FASL deficient isoflurane group, and the number of platform crossing times decreased significantly (P0.05), but there was no significant difference between the control group and the two groups of mice. Conclusion: Isoflurane can induce the cells in the hippocampus of healthy newborn mice by activating FASL-FAS signal pathway. Apoptosis, which can damage the hippocampus and cause the decline of learning and memory ability in the future, can reduce the damage and improve the learning and memory ability in mice that cut off the FASL-FAS signaling pathway, which proves that the FAS pathway is one of the mechanisms of isoflurane damage to the hippocampus of newborn mice.
【學(xué)位授予單位】：復(fù)旦大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R614.2

【共引文獻(xiàn)】

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