荜澄茄素治疗失眠症的药效及相关机制研究  

Efficacy and related mechanism of cubebin in treatment of insomnia

作　　者：王玮 杨世林[1,2] 彭万钱 张壮 欧阳辉 冯育林[1,2] 罗颖颖 何明珍[1,2] WANG Wei;YANG Shi-lin;PENG Wan-qian;ZHANG Zhuang;OUYANG Hui;FENG Yu-lin;LUO Ying-ying;HE Ming-zhen(National Engineering Center of Solid Preparation of Chinese Materia Medical Manufacturing Technology,Jiangxi University of Traditional Chinese Medicine,Nanchang 330006,China;State Key Laboratory of Innovative Drug and Efficient Energy-Saving Pharmaceutical Equipment,Nanchang 330006,China;Nanchang Key Laboratory of Active Ingiedients of Traditional Chinese Medicine and Natural Medicine,Nanchang 330006,China)

机构地区：[1]江西中医药大学中药固体制剂制造技术国家工程研究中心,江西南昌330006 [2]创新药物与高效节能降耗制药设备国家重点实验室,江西南昌330006 [3]南昌市中药与天然药物活性成分研究重点实验室,江西南昌330006

出　　处：《中草药》2022年第21期6750-6758,共9页Chinese Traditional and Herbal Drugs

基　　金：国家重点研发计划课题(2019YFC1712302);中央引导地方科技发展专项项目(20192ZDD02002);江西省教育厅科技计划(201222);中央引导地方科技发展资金项目(20221ZDD02007);江西省大学生创新创业训练计划项目(S202210412081)。

摘　　要：目的探讨荜澄茄素对睡眠剥夺小鼠的药效及潜在作用机制。方法首先研究荜澄茄素对正常小鼠的促睡眠作用,小鼠随机分为对照组、地西泮(2 mg/kg)组和荜澄茄素低、中、高剂量(25、50、100 mg/kg)组,各给药组ig相应药物,记录各组小鼠的入睡率以及睡眠持续时间;免疫组化法检测小鼠脑组织γ-氨基丁酸A受体α1(γ-aminobutyric acid A receptorα1,GABAA)蛋白表达;氟马西尼拮抗实验验证荜澄茄素的作用是否与苯二氮类受体有关。然后研究荜澄茄素对睡眠剥夺小鼠的作用,小鼠分为对照组、模型组、地西泮(2 mg/kg)组和荜澄茄素低、中、高剂量(10、20、40 mg/kg)组,给予药物干预后,采用旷场实验进行行为学测试;LC-MS/MS检测脑组织中神经递质GABA、L-谷氨酸(L-glutamic acid,Glu)、5-羟色胺(5-hydroxytryptamine,5-HT)、5-羟吲哚乙酸(5-hydroxyindoleacetic acid,5-HIAA)的含量;免疫组化及qRT-PCR技术检测小鼠脑组织GABAAα1表达。结果荜澄茄素对正常小鼠的促睡眠作用研究结果显示,荜澄茄素显著增加正常小鼠的入睡率(P<0.01),并显著延长了小鼠的睡眠持续时间(P<0.01),显著增加皮层GABAAα1表达(P<0.05);氟马西尼可以拮抗荜澄茄素对小鼠的促睡眠作用。荜澄茄素对睡眠剥夺小鼠的作用研究结果显示,与对照组比较,模型组小鼠活动量显著增加(P<0.05),脑组织中GABAAα1表达以及GABA、Glu、5-HIAA含量显著降低(P<0.05、0.01);与模型组比较,荜澄茄素组小鼠活动量明显减少(P<0.05、0.01),脑组织中GABAAα1表达以及GABA、Glu、5-HIAA含量显著升高(P<0.05、0.01)。结论荜澄茄素对正常小鼠和睡眠剥夺小鼠均具有促睡眠作用,可能通过调控GABAAα1及脑内神经递质GABA、Glu、5-HIAA的含量而发挥作用。Objective To explore the efficacy and potential mechanism of cubebin in sleep deprived mice.Methods Firstly,sleep promoting effect of cubebin on normal mice was investigated:Mice were randomly divided into control group,diazepam(2 mg/kg)group,cubebin low-,medium-and high-dose(25,50,100 mg/kg)group,each administration group was ig corresponding drugs,sleep rate and sleep duration of rats in each group were recorded;Immunohistochemical staining was used to detectγ-aminobutyric acid A receptorα1(GABAAα1)protein expression in brain tissue of mice;Flumazenil antagonism experiment was used to verify whether the effects of cubebin are related to benzodiazepine receptors.Secondly,effect of cubebin on sleep deprived mice was observed:Mice were divided into control group,model group,diazepam(2 mg/kg)group,cubebin low-,medium-and high-dose(10,20,40 mg/kg)group,drugs were given for intervention,open field test was used for behavioral testing;LC-MS/MS was used to detect the content of neurotransmitters[GABA,L-glutamic acid(Glu),5-hydroxytryptamine(5-HT)and 5-hydroxyindoleacetic acid(5-HIAA)]in brain tissue;Immunohistochemical staining and qRT-PCR were used to detect GABAAα1 expression in brain tissue.Results The results of study on sleep promoting effect of cubebin on normal mice showed that,cubebin significantly increased the sleep rate of normal mice(P<0.01),significantly prolonged the sleep duration of mice(P<0.01),and significantly increased GABAAα1 expression in cerebral cortex(P<0.05);Flumazenil could antagonize the sleep-promoting effect of cubebin.The results of study on effect of cubebin on sleep deprived mice showed that,compared with control group,activity of mice in model group was significantly increased(P<0.05),GABAAα1 expression and GABA,Glu,5-HIAA contents in brain tissue were significantly reduced(P<0.05,0.01);Compared with model group,activity of mice in cubebin group was significantly reduced(P<0.05,0.01),GABAAα1 expression and GABA,Glu,5-HIAA contents in brain tissue were significantly increased(P<0

关 键 词：荜澄茄素 失眠 睡眠剥夺 神经递质 γ-氨基丁酸A受体α1 

分 类 号：R285.5[医药卫生—中药学]