摘要
目的:探讨安寐丹对睡眠剥夺模型大鼠学习记忆能力的影响并探讨机制。方法:50只SD大鼠随机分为空白组、模型组、安寐丹低、高剂量组(4.55、18.18 g·kg^(-1)·d^(-1))和艾司唑仑组(0.09 mg·kg^(-1)·d^(-1)),每组10只,用自制的睡眠剥夺箱进行持续性睡眠剥夺21 d。大鼠的学习记忆水平以Morris水迷宫来检测,免疫荧光检测大鼠海马N-myc下游调控基因2(NDRG2)、胶质纤维酸性蛋白(GFAP)阳性细胞表达量,实时荧光定量聚合酶链式反应(Real-time PCR)检测海马区NDRG2、兴奋性谷氨酸转运体-1(GLT-1)、N-甲基-D-天(门)冬氨酸(NMDA)受体2个调节亚基GluNR2A、GluNR2B mRNA表达。蛋白免疫印迹法(Western blot)检测海马NDRG2、GLT-1蛋白表达。结果:与空白组比较,模型组大鼠上平台的潜伏期、游泳总路程显著增加,穿越目标象限总路程、穿越目标象限时间、穿越平台次数显著减少(P<0.01),海马CA1区NDRG2、GFAP阳性细胞数显著增加(P<0.01),NDRG2、GluNR2B mRNA相对表达量增加,GLT-1、GluNR2A mRNA相对表达量下降,NDRG2蛋白表达水平显著升高(P<0.01),而GLT-1蛋白表达水平显著降低(P<0.01);与模型组比较,安寐丹低、高剂量组大鼠的学习记忆能力显著改善(P<0.01),大鼠海马NDRG2、GFAP阳性细胞数显著减少(P<0.01),NDRG2、GluNR2B mRNA相对表达量明显降低,GLT-1、GluNR2A mRNA相对表达量升高,NDRG2蛋白表达水平明显降低(P<0.05,P<0.01),而GLT-1蛋白表达水平显著升高(P<0.01)。结论:安寐丹可改善睡眠剥夺大鼠的学习记忆能力,其机制可能与调控星形胶质细胞活性影响脑内神经递质水平及突触可塑性有关。
Objective:To explore the effect of Anmeidan(AMD)on the learning and memory ability of sleep-deprived rats and the mechanism.Method:A total of 50 SD rats were randomized into control group,model group,low-dose AMD group(4.55 g·kg^(-1)·d^(-1)),high-dose AMD group(18.18 g·kg^(-1)·d^(-1)),and estazolam group(0.09 mg·kg^(-1)·d^(-1)).Insomnia was induced in rats with the self-made sleep deprivation box(21 days).The learning and memory ability of rats was measured by Morris water maze.Immunofluorescence method was employed to detect the number of cells expressing N-myc downstream-regulated gene 2(NDRG2)and glial fibrillary acidic protein(GFAP)in hippocampus of rats,real-time fluorescent quantitative polymerase chain reaction(Real-time PCR)to determine the mRNA expression of hippocampal NDRG2,glial glutamate transporter-1(GLT-1),and GluNR2A and GluNR2B N-methyl-D-aspartate(NMDA)receptor subunits,and Western blot to examine the protein expression of NDRG2 and GLT-1 in hippocampus.Result:Compared with control group,the model group showed increase in the latency to reach the platform and total swimming distance,significant decrease in the total distance moved in the target quadrant,time in target quadrant,and times of crossing the platform(P<0.01),rise in the number of cells expressing NDRG2 and GFAP in the hippocampal CA1 region(P<0.01)and the mRNA level of NDRG2 and GluNR2B,reduction in the mRNA level of GLT-1 and GluNR2A,elevation in NDRG2 protein expression(P<0.01),and decrease in GLT-1 protein expression(P<0.01).In contrast to the model group,low-dose and high-dose AMD improved the learning and memory levels of sleep-deprived rats(P<0.01),reduced the number of cells expressing NDRG2 and GFAP(P<0.01),significantly decreased the mRNA expression of NDRG2 and GluNR2B,increased the mRNA expression of GLT-1 and GluNR2A,reduced NDRG2 protein level(P<0.05,P<0.01),and raised GLT-1 protein level(P<0.01).Conclusion:AMD can improve the learning and memory ability of sleep-deprived rats.The mechanism is the likelihood that it regulates astrocyte activity,thereby affecting the neurotransmitter level and synaptic plasticity in the brain.
作者
谭云霞
纪可
刘福贵
方洁
刘玲
TAN Yun-xia;JI Ke;LIU Fu-gui;FANG Jie;LIU Ling(Hubei University of Chinese Medicine,Wuhan 430065,China;Hubei Provincial Hospital of Traditional Chinese Medicine,Wuhan 430061,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2022年第7期18-25,共8页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家重点研发计划"中医药现代化研究"重点专项(2018YFC1705602)。
