摘要
目的:观察安寐丹对对氯苯丙氨酸(PCPA)失眠大鼠下丘脑哺乳动物雷帕霉素靶标(mTOR)/真核细胞始动因子4E结合蛋白1(4EBP1)/真核细胞翻译起始因子4E(eIF4E)信号通路的影响。方法:60只SD大鼠随机分为对照组,模型组,安寐丹低、中、高剂量组和褪黑素组,每组10只。除对照组外均采用PCPA腹腔注射建立失眠大鼠模型。运用旷场实验检测大鼠焦虑样行为,Morris水迷宫检测其学习记忆能力,透射电镜观察下丘脑神经元线粒体结构,RT-PCR、Western Blot、免疫荧光检测各组大鼠下丘脑mTOR/4EBP1/eIF4E通路相关基因和蛋白表达情况。结果:与对照组比较,模型组大鼠活动总距离显著增加(P<0.01),中心区停留时间显著减少(P<0.01),上平台潜伏期与游泳总路程均显著延长(P<0.01),穿越平台次数、目标象限停留时间显著减少(P<0.01),且线粒体出现明显肿胀、破损,大部分嵴断裂、溶解,形成空泡样,下丘脑p-mTOR/mTOR、p-4EBP1/4EBP1 mRNA和蛋白表达显著上调(P<0.01),p-eIF4E/eIF4E、PER1、PER2 mRNA和蛋白表达显著下调(P<0.01)。与模型组比较,安寐丹中、高剂量组及褪黑素组大鼠活动总距离显著减少(P<0.05),中心区停留时间显著增加(P<0.05,P<0.01);上平台潜伏期和游泳总路程显著缩短(P<0.05,P<0.01),穿越平台次数和目标象限停留时间显著增加(P<0.05,P<0.01);线粒体损伤减轻;p-mTOR/mTOR、p-4EBP1/4EBP1 mRNA与蛋白表达显著下调(P<0.01),p-eIF4E/eIF4E、PER1、PER2 mRNA与蛋白表达显著上调(P<0.01)。结论:安寐丹改善PCPA失眠大鼠昼夜节律紊乱,其机制可能与调控mTOR/4EBP1/eIF4E信号通路有关。
Objective:To observe the effects of Anmeidan on mammalian target of rapamycin(mTOR)/eukaryotic initiation factor 4E binding protein 1(4EBPI)/eukaryotic translation initiation factor 4E(eIF4E)signaling pathway in hypothalamus of insomnia rats induced by p-chlorophenylalanine(PCPA).Methods:Sixty SD rats were randomly divided into control group,model group,Anmeidan low,medium and high dose groups and melatonin group,with 10 rats in each group.In addition to the control group,the insomnia rat model was established by intraperitoneal injection of PCPA.Open field test was used to detect the anxiety-like behavior of rats,and Morris water maze was used to detect their learning and memory ability.Transmission electron microscopy was used to observe the mitochondrial structure of hypothalamic neurons.RT-PCR,Western Blot and immunofluorescence were used to detect the expression of mTOR/4EBP1/eIF4E pathway-related genes and proteins in hypothalamus of rats in each group.Results:Compared with the control group,the total activity distance of rats in the model group was significantly increased(P<0.01),the residence time in the central area was significantly decreased(P<0.01),the latency of the upper platform and the total swimming distance were significantly prolonged(P<0.01),the number of crossing platforms and the residence time in the target quadrant were decreased(P<0.01),and the mitochondria were significantly obviously swollen and damaged.Most of the cristae were broken and dissolved to form vacuoles,and the mRNA and protein expressions of p-mTOR/mTOR and p-4EBP1/4EBP1 in the hypothalamus were significantly up-regulated(P<0.01).The mRNA and protein expressions of p-eIF4E/eIF4E,PER1 and PER2 were significantly down-regulated(P<0.01).Compared with the model group,the total activity distance of rats in the middle and high dose Anmeidan group and the melatonin group was significantly decreased(P<0.05),and the residence time in the central area was significantly increased(P<0.05,P<0.01).The platform latency and total swimming distance were significantly shortened(P<0.05,P<0.01),and the number of crossing platforms and the target quadrant residence time increased significantly(P<0.05,P<0.01).Mitochondrial damage reduced;the mRNA and protein expressions of p-mTOR/mTOR and p-4EBP1/4EBP1 were significantly down-regulated(P<0.01),and the mRNA and protein expression of p-eIF4E/eIF4E,PER1 and PER2 were significantly up-regulated(P<0.01).Conclusion:Anmeidan can improve circadian rhythm disorder in PCPA insomnia rats,and its mechanism may be related to the regulation of mTOR/4EBP1/eIF4E signaling pathway.
作者
秦庆花
徐波
谢光璟
叶子靖
付顺刚
夏婧
王平
QIN Qinghua;XU Bo;XIE Guangjing;YE Zijing;FU Shungang;XIA Jing;WANG Ping(Elderly Brain Health Traditional Chinese Medicine Protection Technology and New Product Development Engineering Research Center of the Ministry of Education,Hubei University of Chinese Medicine,Wuhan 430065,China)
出处
《中华中医药杂志》
北大核心
2025年第7期3364-3370,共7页
China Journal of Traditional Chinese Medicine and Pharmacy
基金
国家自然科学基金青年科学基金项目(No.82205232)
湖北省自然科学基金青年项目(No.2021CFB256)
武汉市知识创新专项项目(No.2023020201020473)。
