摘要
目的观察黄连温胆汤对氯苯丙氨酸(PCPA)致失眠小鼠的睡眠改善作用及对γ-氨基丁酸(GABA)能系统通路相关受体及递质的影响,探讨其治疗失眠的作用机制。方法将48只雄性KM小鼠随机分为空白组、模型组、地西泮组(2 mg·kg^(-1))及黄连温胆汤低、中、高剂量组(8.45、16.90、33.80 g·kg^(-1))。除空白组外,其余各组小鼠均腹腔注射450 mg·kg^(-1)PCPA(每日1次,连续2 d)建立小鼠失眠模型。造模成功后,各组小鼠灌胃给予相对应药物,每日1次,连续灌胃7 d。观察记录各组小鼠的活动次数、体质量及戊巴比妥钠阈上剂量诱导的睡眠潜伏期、睡眠时间;采用ELISA法测定小鼠脑组织中谷氨酸(GLU)、GABA含量,计算GLU/GABA比值;采用免疫组织化学法测定脑组织中谷氨酸脱羧酶65/67(GAD65/67)、γ-氨基丁酸转运蛋白亚型1(GAT-1)、γ-氨基丁酸A受体α1亚型(GABA ARα1)、γ-氨基丁酸A受体γ-2亚型(GABA ARγ-2)及Na+-K+-Cl-协同转运蛋白1(NKCC1)的表达情况;采用Western Blot法测定小鼠脑组织中GAD65/67、GABA ARα1及NKCC1蛋白的表达情况。结果与空白组比较,模型组小鼠戊巴比妥钠阀上剂量诱导的睡眠潜伏期显著延长(P<0.01),睡眠时间显著缩短(P<0.01),活动次数显著增加(P<0.01),体质量显著降低(P<0.01),脑组织中的GABA含量明显下降(P<0.05),GLU含量及GLU/GABA比值显著升高(P<0.01),脑组织中GAD65/67、GABA ARα1蛋白表达显著下调(P<0.01),NKCC1蛋白表达显著上调(P<0.01)。与模型组比较,黄连温胆汤可有效缩短戊巴比妥钠阀上剂量诱导的小鼠睡眠潜伏期(P<0.05,P<0.01),并延长小鼠睡眠时间(P<0.01),减少造模后小鼠活动次数(P<0.01),使体质量显著上升(P<0.05,P<0.01),升高脑组织中的GABA含量(P<0.01),降低GLU含量及GLU/GABA比值(P<0.01),并显著上调脑组织中GAD65/67、GAT-1、GABA ARα1、GABA ARγ-2蛋白表达(P<0.01),下调NKCC1蛋白表达(P<0.01)。结论黄连温胆汤具有较好的镇静催眠
Objective To observe the effects of Huanglian Wendan Decoction on the sleep improvement of parachlorophenylalanine(PCPA)-induced insomnia mice and the effects on the receptors and neurotransmitters related to the gamma-aminobutyric acid(GABA)pathway,and to explore its mechanism of action in the treatment of insomnia.Methods Forty-eight male KM mice were randomly divided into the blank group,the model group,the diazepam group(2 mg·kg^(-1))and the low-,medium-and high-dose groups(8.45,16.90 and 33.80 g·kg^(-1))of Huanglian Wendan Decoction.Except for the blank group,mice in all groups were injected intraperitoneally with 450 mg·kg^(-1)PCPA(once daily for consecutive 2 days)to establish a mouse insomnia model.After successful modeling,mice in each group were given corresponding drugs by gavage once a day for consecutive 7 days.The number of activities,body mass,sleep latency and sleep duration induced by suprathreshold dose of pentobarbital sodium were observed and recorded;the contents of glutamic acid(GLU)and gamma-aminobutyric acid(GABA)in brain tissue were determined by ELISA,and the ratio of GLU/GABA was calculated,the expression of glutamate decarboxylase 65/67(GAD65/67),gamma-aminobutyric acid transporter subtype-1(GAT-1),gamma-aminobutyric acid A receptor subtypeα1(GABA ARα1),gamma-aminobutyric acid A receptor subtypeγ-2(GABA ARγ-2)and Na^(+)-K^(+)-Cl-cotransport protein 1(NKCC1)in brain tissues were determined by ELISA,and the GLU/GABA ratio were calculated;the protein expressions of GAD65/67,GABA ARα1 and NKCC1 in mouse brain tissues were determined by Western Blot.Results Compared with the blank group,the pentobarbital sodium-induced sleep latency of mice in the model group was significantly prolonged(P<0.01),the sleep duration was significantly shortened(P<0.01),the frequency of activity was significantly increased(P<0.01),the body mass was significantly reduced(P<0.01),the GABA content in brain tissue was significantly decreased(P<0.05),the GLU content and GLU/GABA ratio were significantly incre
作者
胡莹
李亮
戴卫波
王家赐
陈巧玲
郭文贤
林慧
谢为
李青
HU Ying;LI Liang;DAI Weibo;WANG Jiaci;CHEN Qiaoling;GUO Wenxian;LIN Hui;XIE Wei;LI Qing(Zhongshan Hospital of Traditional Chinese Medicine,Zhongshan 528400 Guangdong,China)
出处
《中药新药与临床药理》
CAS
CSCD
北大核心
2022年第8期1009-1016,共8页
Traditional Chinese Drug Research and Clinical Pharmacology
基金
中山市社会公益科技研究重大项目(2019B1016)。
