围产期服用OMEGA-3多不饱和脂肪酸改善七氟烷导致的新生大鼠神经退行性病变和记忆损害

发布时间：2018-06-07 11:39

本文选题：七氟烷 + OMEGA-3不饱和脂肪酸　；参考：《复旦大学》2014年硕士论文

【摘要】：近年来许多动物实验发现,新生动物吸入七氟烷等麻醉气体能导致神经细胞的凋亡,影响神经细胞的增生和分化,从而损害神经认知功能。OMEGA-3多不饱和脂肪酸作为一类重要的不饱和脂肪酸,在脑内含量极为丰富,在维持脑的结构和功能上具有极为重要的作用。以往的实验发现,OMEGA-3多不饱和脂肪酸具有抗凋亡,抗氧化和改善神经发生的作用。本实验将观察七氟烷是否能对未成熟脑产生明显的神经毒性作用,且围产期服用含丰富OMEGA-3多不饱和脂肪酸的鱼油是否能改善七氟烷导致新生大鼠的神经毒性和认知损害。斯普拉格-杜勒(SD)孕鼠(每组3只)从怀孕第二天(怀孕期)开始到分娩后14天(哺乳期)服用含鱼油或不含鱼油的食物。其新生大鼠在出生后第7天给予吸入麻醉药七氟烷干预(母鼠没有鱼油干预,新生鼠也没有进行七氟烷干预的为对照组)。每组的新生大鼠在麻醉刚开始的时候和麻醉后持续3天内定时腹腔注射BrdU,以评估海马区的神经增生。其后每组新生大鼠在指定的时间分别取脑组织和灌注固定,冰冻切片。各组在麻醉后3小时(每组3只)用流式细胞术的方法分析各组细胞活性氧(ROS)的相对水平；在麻醉后18小时进行用免疫蛋白印记的方法和免疫荧光的方法检测脑组织内活化型半胱天冬酶-3 (cleaved caspase-3)的变化；在麻醉后3天观察各组BrdU的含量。剩余新生大鼠(每组9只)则分别在出生后5周,9周和10周的时候行莫里斯(Morris)水迷宫空间学习记忆、场景恐惧实验和莫里斯水迷宫强化记忆实验。实验发现,3%七氟烷6小时持续吸入能导致新生大鼠大脑的丘脑区,顶叶皮层区活化型半胱天冬酶-3(cleaved caspase-3)明显的增多,但在海马区却没看到明显差异。且在皮层区中,我们进而发现七氟烷能导致细胞活性氧(ROS)的增多。七氟烷还能降低海马齿状回区的BrdU标记阳性的神经前体细胞。然而,我们发现围产期服用鱼油能明显降低新生大鼠皮层区活化型半胱天冬酶-3 (cleaved caspase-3)和细胞活性氧(ROS),且改善海马区齿状回BrdU标记阳性的神经前体细胞。在行为学实验中,虽然各组新生鼠成年后在莫里斯水迷宫空间学习记忆和莫里斯水迷宫强化记忆实验中的早期长期记忆了测验中没有明显差异,但是我们发现七氟烷能明显损伤新生大鼠成年后对场景恐惧的即刻反应、莫里斯水迷宫强化记忆实验中的工作记和短期记忆,并且鱼油能明显改善七氟烷造成的学习记忆损伤。我们的实验发现了新生鼠吸入七氟烷能造成神经细胞氧化,凋亡和降低神经新生,最终导致成年时场景恐惧的即刻反应、莫里斯水迷宫强化记忆实验中的工作记和短期记忆的损害。然而围产期服用鱼油能减轻七氟烷的造成的神经毒性作用。
[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.
【学位授予单位】：复旦大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R614

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