大麻二酚配伍褪黑素改善大、小鼠失眠和昼夜节律紊乱作用及机制  

Curative effects and mechanism of cannabidiol-melatonin formula on insomnia and circadian rhythm disturbance in mice and rats

作　　者：刘影 李劲草 薛瑞 张杨 李硕 董华进 傅风华[1] 张有志 LIU Ying;LI Jin-cao;XUE Rui;ZHANG Yang;LI Shuo;DONG Hua-jin;FU Feng-hua;ZHANG You-zhi(College of Pharmacy,Yantai University,Yantai 264005,China;State Key Laboratory of Antidrugs and Toxicology,Institute of Pharmacology and Toxicology,Academy of Military Medical Sciences,Beijing 100850,China)

机构地区：[1]烟台大学药学院,山东烟台264005 [2]军事科学院军事医学研究院毒物药物研究所,抗毒药物与毒理学国家重点实验室,北京100850

出　　处：《中国药理学与毒理学杂志》2023年第2期81-90,共10页Chinese Journal of Pharmacology and Toxicology

基　　金：国家科技重大专项(2018ZX09739-008)。

摘　　要：目的评价大麻二酚(CBD)配伍褪黑素(MLT)(CBD-MLT复方)对生物节律紊乱与失眠的调节作用及机制。方法①雄性ICR小鼠随机分为正常对照组、CBD(5,10和20 mg·kg^(-1))组、MLT(2.5,5,10和20 mg·kg^(-1))组及CBD-MLT复方〔1.25+2.5,2.5+2.5,1.25+5和(2.5+5)mg·kg^(-1)〕组。ig给药后ip给予戊巴比妥钠(48 mg·kg^(-1))进行翻正反射实验,考察各组药物对正常小鼠睡眠持续时间的影响。②雄性SD大鼠进行植入子手术,随机分为正常对照组、阳性药物地西泮(DZP 3 mg·kg^(-1))组和CBD-MLT复方(10+10)mg·kg^(-1)组。ig给药后进行无线生理信号遥测与睡眠分析,考察各组药物对正常大鼠睡眠时间和睡眠结构的影响。③雄性ICR小鼠随机分为正常对照组、模型组、模型+DZP(3 mg·kg^(-1))组及模型+CBD-MLT复方〔5+5,10+10和(20+20)mg·kg^(-1)〕组,连续5 d ip给予对氯苯丙氨酸(PCPA)诱导失眠,第6天ig给药后ip给予戊巴比妥钠(48 mg·kg^(-1))进行翻正反射实验,考察CBD-MLT复方对入睡潜伏期和睡眠持续时间的改善作用;借助自发活动视频分析系统对失眠小鼠活动总路程和活动速度进行分析,通过活动-静息节律的变化考察CBDMLT复方(20+20)mg·kg^(-1)对小鼠昼夜节律的调节作用;生化试剂盒检测小鼠血清超氧化物歧化酶(SOD)和过氧化氢酶(CAT)活性及丙二醛(MDA)和谷胱甘肽(GSH)含量;Western印迹法检测小鼠下丘脑视交叉上核脑肌类芳烃受体核转位样蛋白1(Bmal1)、时钟节律调节蛋白(Clock)、节律周期调节蛋白2(Per2)、隐花色素节律调节蛋白2(Cry2)和5-羟色胺1A受体(5HT_(1A)R)蛋白表达水平。结果①与正常对照组比较,CBD或MLT协同戊巴比妥钠延长小鼠睡眠时间的最低有效剂量分别为20 mg·kg^(-1)(P<0.01)和10 mg·kg^(-1)(P<0.05),而CBD-MLT复方的最低有效剂量为(1.25+2.5)mg·kg^(-1)(P<0.01)。②大鼠脑电睡眠分析结果表明,与正常对照组比较,CBD-MLT复方(10+10)mg·kg^(-1)可增加大鼠睡眠总时OBJECTIVE To evaluate the effects of cannabidiol-melatonin(CBD-MLT)formula on insomnia and circadian rhythm disturbance in mice and explore its possible mechanism.METHODS①Male ICR mice were randomly divided into the normal control,CBD(5,10,20 mg·kg^(-1)),MLT(2.5,5,10,20 mg·kg^(-1))and CBD-MLT[1.25+2.5,2.5+2.5,1.25+5,(2.5+5)mg·kg^(-1)]group.After ig administra⁃tion of corresponding drugs,pentobarbital sodium(48 mg·kg^(-1))was ip injected for righting reflex test to investigate the effect of each drug on the sleep duration of normal mice.②Male SD rats were randomly divided into the normal control,diazepam(DZP,3 mg·kg^(-1))and CBD-MLT(10+10)mg·kg^(-1)groups.After ig administration of corresponding drugs(20 mL·kg^(-1)),wireless physiological signal telemetry and sleep analysis were performed to investigate the effects of the drug on sleep time and sleep structure of normal rats.③Male ICR mice were randomly divided into the normal control,model,model+DZP(3 mg·kg^(-1))and model+CBD-MLT[5+5,10+10,(20+20)mg·kg^(-1)]formula groups.Insomnia was induced by ip injection of P-chlorophenylalanine(PCPA)for 5 consecutive days.On the 6th day,pentobar⁃bital sodium(48 mg·kg^(-1))was ip injected for righting reflex test to investigate the effect of CBD-MLT on improving sleep latency and sleep duration.Using the spontaneous activity video analysis system,the total distance and speed of mice were analyzed,while the regulation effect of CBD-MLT(20+20)mg·kg^(-1)on circadian rhythm of model mice was investigated through the changes of activity-rest rhythm.The serum oxidative stress indicators superoxide dismutase(SOD),malondialdehyde(MDA),catalase(CAT)and glutathione(GSH)were detected by biochemical reaction kit.The core circadian clock proteins brain and muscle Arnt-like protein^(-1)(Bmal1),circadian locomoter output cycles protein kaput(Clock),period circadian regulator 2(Per2),cryptochrome circadian regulator 2(Cry2),and 5-hydroxytryptamine 1A receptor(5HT_(1A)R)in suprachiasmatic nucleus(SCN)were tested by Wester

关 键 词：大麻二酚 褪黑素 生物节律 失眠 氧化还原 

分 类 号：R964[医药卫生—药理学]