摘要
背景:稳定的动物失眠模型是研究失眠机制和抗失眠药物研发的重要工具。化学法结合物理方法构建失眠模型为动物失眠模型提供了良好的造模方法。目的:比较化学方法(对氯苯丙氨酸法)、物理方法(水环境睡眠剥夺法)和化学联合物理方法(对氯苯丙氨酸联合水环境睡眠剥夺法)构建小鼠失眠模型,探索最佳的造模方法。方法:将C57小鼠分别按照雄、雌性别随机分成空白组(不做处理)、对氯苯丙氨酸组、水平台组、对氯苯丙氨酸+水平台组,每组12只(雌、雄各半)。对氯苯丙氨酸组小鼠每天腹腔注射400 mg/kg对氯苯丙氨酸混悬液,连续3 d;水平台组小鼠连续3 d放入水环境睡眠剥夺平台中;对氯苯丙氨酸+水平台组小鼠放入睡眠剥夺平台中,且连续3 d腹腔注射400 mg/kg对氯苯丙氨酸混悬液。造模3 d后,对小鼠进行旷场实验、尼氏染色观察下丘脑及海马区神经元数量,酶联免疫吸附法检测血清中5-羟色胺、γ-氨基丁酸、多巴胺水平。结果与结论:①与空白组小鼠比较,3种模型组雄性小鼠的总路程、中心进入次数、平均速度和站立次数均呈上升趋势,其中对氯苯丙氨酸+水平台组变化最为明显(P<0.05或P<0.01);3种模型组雌性小鼠总路程、中心进入次数、平均速度和站立次数均呈上升趋势;②3种造模方法都可以减少小鼠下丘脑、海马区神经元数量,其中对氯苯丙氨酸+水平台组雌性小鼠神经元数量减少最为明显,神经元损伤最为严重;③与空白组相应性别小鼠比较,对氯苯丙氨酸+水平台组雌性小鼠血清3项指标变化更显著,5-羟色胺、γ-氨基丁酸显著降低(P<0.05),多巴胺水平显著升高(P<0.01)。结果说明:对氯苯丙氨酸联合水平台环境法建立的失眠模型对雌性小鼠相对更可靠、稳定,能为失眠研究提供较为理想的动物模型。
BACKGROUND:Stable animal insomnia models serve as crucial tools for investigating insomnia mechanisms and developing anti-insomnia drugs.The combination of chemical and physical methods provides an effective approach for establishing animal insomnia models.OBJECTIVE:To compare chemical methods(p-chlorophenylalanine),physical methods(water environment sleep deprivation method),and combined methods(p-chlorophenylalanine combined with water environment sleep deprivation method)for constructing insomnia mouse models,thereby exploring the optimal modeling method.METHODS:C57 mice were randomly divided by sex into the following groups:control(untreated),p-chlorophenylalanine,water platform,and p-chlorophenylalanine+water platform,with 12 mice per group(equal numbers of males and females).Mice in the p-chlorophenylalanine group received intraperitoneal injections of 400 mg/kg p-chlorophenylalanine suspension daily for 3 consecutive days;those in the water platform group were placed in a water environment sleep deprivation platform for 3 consecutive days;and those in the p-chlorophenylalanine+water platform group were placed in the sleep deprivation platform and received intraperitoneal injections of 400 mg/kg p-chlorophenylalanine suspension for 3 consecutive days.After 3 days of modeling,mice were subjected to the open field test,and the number of neurons in the hypothalamus and hippocampus was examined via Nissl staining.Serum levels of 5-hydroxytryptamine,gamma-aminobutyric acid,and dopamine were measured using enzyme-linked immunosorbent assay.RESULTS AND CONCLUSION:(1)Compared with the control group,male mice in the three model groups exhibited an upward trend in total distance,number of entries into the central area,average speed,and number of stand-up episodes.The most pronounced changes were observed in the p-chlorophenylalanine+water platform group(P<0.05 or P<0.01).Female mice in the three model groups also showed an upward trend in total distance,number of entries into the central area,average speed,and number of stand-up episodes.(2)All three modeling methods could reduce the number of neurons in the mouse hypothalamus and hippocampus,with the highest reduction and most severe neuronal damage observed in female mice in the p-chlorophenylalanine+water platform group.(3)Compared with female mice in the control group,changes in three serum indicators were more pronounced in the female mice in the p-chlorophenylalanine+water platform group.5-Hydroxytryptamine and gamma-aminobutyric acid levels were significantly reduced(P<0.05),while dopamine levels were significantly elevated(P<0.01).To conclude,the insomnia model established in female mice using p-chlorophenylalanine combined with the water platform is relatively more reliable and stable,providing an ideal animal model for insomnia research.
作者
李非凡
张怡博
王静
朱金墙
郑文科
Li Feifan;Zhang Yibo;Wang Jing;Zhu Jinqiang;Zheng Wenke(Institute of Traditional Chinese Medicine,Tianjin University of Traditional Chinese Medicine,Tianjin 301617,China)
出处
《中国组织工程研究》
北大核心
2026年第18期4685-4693,共9页
Chinese Journal of Tissue Engineering Research
基金
天津市卫生健康委员会中医中西医结合科研课题(2023079),项目负责人:朱金墙
黑龙江省“揭榜挂帅”科技攻关(2023ZXJ02C01),项目负责人:郑文科。
关键词
失眠
动物模型
对氯苯丙氨酸
水环境睡眠剥夺
多巴胺
小鼠
insomnia
animal model
p-chlorophenylalanine
water environment sleep deprivation
dopamine
mouse
