建立小鼠学习记忆损伤模型的方法学研究
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摘要
目的通过避暗和Morris水迷宫实验研究建立小鼠学习记忆损伤模型的有效方法。方法分别在小鼠避暗电击训练之前20 min腹腔注射东莨菪碱1 mg/kg体质量,电击训练结束之后立即皮下注射亚硝酸钠90 mg/kg和下次电击测试之前15 min给小鼠灌服35%乙醇0.1 mL/10 g,建立记忆获得、记忆巩固和记忆再现障碍模型,测定各组小鼠在电击训练后6 h、24 h、30 h、48 h后的避暗潜伏期、错误次数和探头次数等多项指标,评价三种记忆障碍模型方法各指标的敏感性和最适测试时间。小鼠分别腹腔注射东莨菪碱1 mg/kg和灌服35%乙醇0.1 mL/10 g造成学习记忆损伤,通过Morris水迷宫定位航行试验和空间探索试验用于测试实验动物对空间位置感和方向感(空间定位)的学习记忆能力,建立空间学习记忆障碍小鼠模型的评价方法。结果避暗实验显示:和空白组比较,东莨菪碱造成记忆获得障碍模型小鼠在电击训练之后6 h、24 h避暗潜伏期明显缩短,错误次数也明显增多(P<0.01、P<0.05);亚硝酸钠造成记忆巩固障碍模型小鼠在电击训练之后30 h、48 h避暗潜伏期明显缩短,错误次数明显增多(P<0.01、P<0.05);乙醇造成的记忆再现障碍模型小鼠在电击训练24 h后即在下次测试之前15 min给小鼠灌服乙醇,可见小鼠避暗潜伏期明显缩短,错误次数明显增多(P<0.05、P<0.01)。和空白组比较,三种药物造成的记忆损伤模型小鼠探头次数均在30 h、48 h增加(P<0.01)。Morris水迷宫实验显示:与空白组相比,东莨菪碱小鼠模型组定位航行逃避潜伏期明显延长而主动探索穿越平台的次数明显减少(P<0.01或P<0.05);乙醇模型组无明显差异(P>0.05)。结论避暗实验中使用东莨菪碱、亚硝酸钠构建小鼠记忆获得障碍和记忆巩固模型时应在造模后24 h和30 h检测相应指标;乙醇造成的记忆再现障碍模型,应在电击训练24 h后,在下次测试之前15 min给小鼠灌服乙醇后检测,指标以避暗潜伏期、错误次数和探头次数为宜。Morris水迷宫实验提示东莨菪碱能够损害小鼠空间学习记忆,可以作为构建小鼠空间学习记忆障碍模型的药物,乙醇并不能建立小鼠的空间记忆障碍模型。
Objective To establish an effective method for learning and memory impairment in mice by avoiding darkness and Morris water maze experiments. Method Scopolamine 1 mg/kg was intraperitoneally injected 20 min before the mice were protected from dark electric shock. Immediately after the end of the electric shock training, sodium nitrite 90 mg/kg was administered subcutaneously and the mice were given 35% ethanol 0.1 mL/10 g 15 minutes before the next shock test., establish a model of memory acquisition, memory consolidation and memory reconstruction disorder, and determine the indicators of avoidance darkness, number of errors and number of probes after 6 h, 24 h, 30 h, 48 h after shock training in each group. The sensitivity of the indicators of the memory impairment model method and the optimal test time. The mice were intraperitoneally injected with scopolamine 1 mg/kg and 35% ethanol 0.1 mL/10 g to cause learning and memory impairment. The Morris water maze positioning navigation test and space exploration test were used to test the spatial orientation and direction of the experimental animals(spatial positioning), the learning and memory ability, the establishment of a spatial learning and memory impairment mouse model evaluation method. Result The dark-avoidance experiment showed that compared with the blank group, the mice with memory impairment caused by scopolamine significantly shortened the dark-avoidance period at 6 h and 24 h after electric shock training, and the number of errors increased significantly(P<0.01, P<0.05). The model of memory consolidation disorder caused by sodium was significantly shortened at 30 h and 48 h after shock training, and the number of errors was significantly increased(P<0.01, P<0.05). The memory-rehabilitation model mice caused by ethanol were after 24 h of electric shock training. The mice were given ethanol at 15 min before the next test, and the dark latency of the mice was significantly shortened, and the number of errors was significantly increased(P<0.05, P<0.01). Compared with the blank group, the number of probes in the memory impairment model caused by the three drugs increased at 30 h and 48 h(P<0.01). In the Morris water maze experiment, compared with the blank group, the scoring escape latency was significantly prolonged in the scopolamine mouse model group and the number of active exploration through the platform was significantly reduced(P<0.01 or P<0.05), and there was no significant difference in the ethanol model group(P>0.05). Conclusion When using scopolamine and sodium nitrite to construct memory impairment and memory consolidation model in mice, the corresponding indicators should be detected at 24 h and 30 h after modeling. The model of memory reproduction disorder caused by ethanol should be 24 h after electric shock training. The mice were tested with ethanol after 15 min before the next test. The indicators were to avoid dark latency, number of errors and number of probes. Morris water maze experiments suggest that scopolamine can damage spatial learning and memory in mice, and can be used as a drug to construct a model of spatial learning and memory impairment in mice. Ethanol cannot establish a model of spatial memory impairment in mice.
Objective To establish an effective method for learning and memory impairment in mice by avoiding darkness and Morris water maze experiments. Method Scopolamine 1 mg/kg was intraperitoneally injected 20 min before the mice were protected from dark electric shock. Immediately after the end of the electric shock training, sodium nitrite 90 mg/kg was administered subcutaneously and the mice were given 35% ethanol 0.1 mL/10 g 15 minutes before the next shock test., establish a model of memory acquisition, memory consolidation and memory reconstruction disorder, and determine the indicators of avoidance darkness, number of errors and number of probes after 6 h, 24 h, 30 h, 48 h after shock training in each group. The sensitivity of the indicators of the memory impairment model method and the optimal test time. The mice were intraperitoneally injected with scopolamine 1 mg/kg and 35% ethanol 0.1 mL/10 g to cause learning and memory impairment. The Morris water maze positioning navigation test and space exploration test were used to test the spatial orientation and direction of the experimental animals(spatial positioning), the learning and memory ability, the establishment of a spatial learning and memory impairment mouse model evaluation method. Result The dark-avoidance experiment showed that compared with the blank group, the mice with memory impairment caused by scopolamine significantly shortened the dark-avoidance period at 6 h and 24 h after electric shock training, and the number of errors increased significantly(P<0.01, P<0.05). The model of memory consolidation disorder caused by sodium was significantly shortened at 30 h and 48 h after shock training, and the number of errors was significantly increased(P<0.01, P<0.05). The memory-rehabilitation model mice caused by ethanol were after 24 h of electric shock training. The mice were given ethanol at 15 min before the next test, and the dark latency of the mice was significantly shortened, and the number of errors was significantly increased(P<0.05, P<0.01). Compared with the blank group, the number of probes in the memory impairment model caused by the three drugs increased at 30 h and 48 h(P<0.01). In the Morris water maze experiment, compared with the blank group, the scoring escape latency was significantly prolonged in the scopolamine mouse model group and the number of active exploration through the platform was significantly reduced(P<0.01 or P<0.05), and there was no significant difference in the ethanol model group(P>0.05). Conclusion When using scopolamine and sodium nitrite to construct memory impairment and memory consolidation model in mice, the corresponding indicators should be detected at 24 h and 30 h after modeling. The model of memory reproduction disorder caused by ethanol should be 24 h after electric shock training. The mice were tested with ethanol after 15 min before the next test. The indicators were to avoid dark latency, number of errors and number of probes. Morris water maze experiments suggest that scopolamine can damage spatial learning and memory in mice, and can be used as a drug to construct a model of spatial learning and memory impairment in mice. Ethanol cannot establish a model of spatial memory impairment in mice.
引文
[1] 袁树民,曹兴水,高翔,等.川芎嗪对痴呆小鼠模型学习记忆能力的影响[J].中国比较医学杂志,2010,20(5):46-49.
[2] 李惠萍,吴钢,莫玉兰,等.氯胺酮影响小鼠避暗潜伏期和错误次数的机制[J].广西医学,2006,(9):1423-1424.
[3] 张永亮,陈海龙,王婷梅,等.木犀草素对东莨菪碱所致大鼠学习记忆障碍的改善作用[J].化学与生物工程,2017,34(2):33-37+49.
[4] 白杨,辛随成.阿尔兹海默病动物模型的研究进展[J].实验动物科学,2013,30(6):61-65+53.
[5] 侯悦,吴春福,何祥,等.氟哌啶醇对小鼠在避暗实验中学习记忆获得、巩固和再现过程的影响[J].中国临床康复,2006,(34):99-102.
[6] 仇淑君,王忠良,李广意,等.丹参对AD模型小鼠学习记忆及脑内和血清内乙酰胆碱酯酶含量的影响[J].实验动物科学,2014,31(4):11-13+31.
[7] 侯悦,吴春福,何祥,等.氯氮平、奥氮平对小鼠在避暗实验中学习记忆获得、巩固和再现过程的影响[J].中国临床康复,2006,(38):61-64.
[2] 李惠萍,吴钢,莫玉兰,等.氯胺酮影响小鼠避暗潜伏期和错误次数的机制[J].广西医学,2006,(9):1423-1424.
[3] 张永亮,陈海龙,王婷梅,等.木犀草素对东莨菪碱所致大鼠学习记忆障碍的改善作用[J].化学与生物工程,2017,34(2):33-37+49.
[4] 白杨,辛随成.阿尔兹海默病动物模型的研究进展[J].实验动物科学,2013,30(6):61-65+53.
[5] 侯悦,吴春福,何祥,等.氟哌啶醇对小鼠在避暗实验中学习记忆获得、巩固和再现过程的影响[J].中国临床康复,2006,(34):99-102.
[6] 仇淑君,王忠良,李广意,等.丹参对AD模型小鼠学习记忆及脑内和血清内乙酰胆碱酯酶含量的影响[J].实验动物科学,2014,31(4):11-13+31.
[7] 侯悦,吴春福,何祥,等.氯氮平、奥氮平对小鼠在避暗实验中学习记忆获得、巩固和再现过程的影响[J].中国临床康复,2006,(38):61-64.