本文選題：七氟烷 + OMEGA-3不飽和脂肪酸　； 參考：《復旦大學》2014年碩士論文

【摘要】：近年來許多動物實驗發(fā)現(xiàn),新生動物吸入七氟烷等麻醉氣體能導致神經細胞的凋亡,影響神經細胞的增生和分化,從而損害神經認知功能。OMEGA-3多不飽和脂肪酸作為一類重要的不飽和脂肪酸,在腦內含量極為豐富,在維持腦的結構和功能上具有極為重要的作用。以往的實驗發(fā)現(xiàn),OMEGA-3多不飽和脂肪酸具有抗凋亡,抗氧化和改善神經發(fā)生的作用。本實驗將觀察七氟烷是否能對未成熟腦產生明顯的神經毒性作用,且圍產期服用含豐富OMEGA-3多不飽和脂肪酸的魚油是否能改善七氟烷導致新生大鼠的神經毒性和認知損害。斯普拉格-杜勒(SD)孕鼠(每組3只)從懷孕第二天(懷孕期)開始到分娩后14天(哺乳期)服用含魚油或不含魚油的食物。其新生大鼠在出生后第7天給予吸入麻醉藥七氟烷干預(母鼠沒有魚油干預,新生鼠也沒有進行七氟烷干預的為對照組)。每組的新生大鼠在麻醉剛開始的時候和麻醉后持續(xù)3天內定時腹腔注射BrdU,以評估海馬區(qū)的神經增生。其后每組新生大鼠在指定的時間分別取腦組織和灌注固定,冰凍切片。各組在麻醉后3小時(每組3只)用流式細胞術的方法分析各組細胞活性氧(ROS)的相對水平；在麻醉后18小時進行用免疫蛋白印記的方法和免疫熒光的方法檢測腦組織內活化型半胱天冬酶-3 (cleaved caspase-3)的變化；在麻醉后3天觀察各組BrdU的含量。剩余新生大鼠(每組9只)則分別在出生后5周,9周和10周的時候行莫里斯(Morris)水迷宮空間學習記憶、場景恐懼實驗和莫里斯水迷宮強化記憶實驗。實驗發(fā)現(xiàn),3%七氟烷6小時持續(xù)吸入能導致新生大鼠大腦的丘腦區(qū),頂葉皮層區(qū)活化型半胱天冬酶-3(cleaved caspase-3)明顯的增多,但在海馬區(qū)卻沒看到明顯差異。且在皮層區(qū)中,我們進而發(fā)現(xiàn)七氟烷能導致細胞活性氧(ROS)的增多。七氟烷還能降低海馬齒狀回區(qū)的BrdU標記陽性的神經前體細胞。然而,我們發(fā)現(xiàn)圍產期服用魚油能明顯降低新生大鼠皮層區(qū)活化型半胱天冬酶-3 (cleaved caspase-3)和細胞活性氧(ROS),且改善海馬區(qū)齒狀回BrdU標記陽性的神經前體細胞。在行為學實驗中,雖然各組新生鼠成年后在莫里斯水迷宮空間學習記憶和莫里斯水迷宮強化記憶實驗中的早期長期記憶了測驗中沒有明顯差異,但是我們發(fā)現(xiàn)七氟烷能明顯損傷新生大鼠成年后對場景恐懼的即刻反應、莫里斯水迷宮強化記憶實驗中的工作記和短期記憶,并且魚油能明顯改善七氟烷造成的學習記憶損傷。我們的實驗發(fā)現(xiàn)了新生鼠吸入七氟烷能造成神經細胞氧化,凋亡和降低神經新生,最終導致成年時場景恐懼的即刻反應、莫里斯水迷宮強化記憶實驗中的工作記和短期記憶的損害。然而圍產期服用魚油能減輕七氟烷的造成的神經毒性作用。
[Abstract]:In recent years, many animal experiments have found that inhaling sevoflurane and other anesthetic gases in newborn animals can lead to apoptosis of nerve cells and affect the proliferation and differentiation of nerve cells. As a kind of important unsaturated fatty acids, OMEGA-3 polyunsaturated fatty acids are abundant in the brain and play an important role in maintaining the structure and function of the brain. Previous experiments have found that OMEGA-3 polyunsaturated fatty acids have the effects of anti-apoptosis, anti-oxidation and improving neurogenesis. The aim of this study was to investigate whether sevoflurane could produce obvious neurotoxicity in immature brain and whether perinatal fish oil rich in OMEGA-3 polyunsaturated fatty acids could improve neurotoxicity and cognitive impairment in neonatal rats induced by sevoflurane. SD) pregnant mice (3 in each group) were given fish oil or no fish oil from the second day of pregnancy (pregnancy) until 14 days after delivery (lactation). The newborn rats were given sevoflurane on the 7th day after birth (the female rats did not interfere with fish oil and the newborn rats did not take the sevoflurane intervention as the control group). The neonatal rats in each group were injected intraperitoneally regularly with BrdU at the beginning of anesthesia and within 3 days after anesthesia to evaluate the hippocampal neurogenesis. After that, brain tissue and perfusion fixation were taken from each group of neonatal rats at a specified time, and frozen sections were taken. The relative levels of reactive oxygen species (Ros) in each group were analyzed by flow cytometry 3 hours after anesthesia. The changes of activated caspase-3 (caspase-3) in brain tissue were detected by immunoglobulin imprinting and immunofluorescence at 18 hours after anesthesia, and the contents of BrdU in each group were observed 3 days after anesthesia. The remaining newborn rats (9 rats in each group) were treated with Morris's water maze spatial learning memory, scene fear test and Morris water maze reinforcement memory experiment at 5 weeks and 10 weeks after birth respectively. It was found that continuous inhalation of 3% sevoflurane for 6 hours resulted in an increase of activated caspase-3 in the thalamus and parietal cortex of neonatal rats, but there was no significant difference in hippocampal area. In the cortical area, we further found that sevoflurane can lead to the increase of reactive oxygen species (Ros). Sevoflurane also reduced BrdU labeled neural precursor cells in dentate gyrus. However, we found that perinatal administration of fish oil could significantly reduce the activation of caspase-3 (caspase-3) and reactive oxygen species (Ros), and improve the BrdU labeled neural precursor cells in dentate gyrus of neonatal rats. In the behavioral experiment, although there was no significant difference in early long-term memory in Morris water maze spatial learning and memory in Morris water maze, there was no significant difference in early long-term memory between each group of newborn rats in adulthood. However, we found that sevoflurane can significantly damage the immediate response to scene fear in newborn rats in adulthood, work memory and short-term memory in Morris water maze, and fish oil can significantly improve learning and memory impairment caused by sevoflurane. Our experiment found that sevoflurane inhalation in newborn rats resulted in the oxidation of nerve cells, apoptosis and reduced neurogenesis, leading to an immediate response to fear in adulthood. Morris Water Labyrinth enhanced memory experiment in work memory and short-term memory impairment. However, perinatal use of fish oil alleviates the neurotoxic effects of sevoflurane.
【學位授予單位】：復旦大學
【學位級別】：碩士
【學位授予年份】：2014
【分類號】：R614

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相關期刊論文 前1條

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本文編號：1991046