基于神经递质和5-HT_(1A)/Gα_(i/o)/cAMP信号通路研究黄连温胆汤对失眠大鼠的治疗作用及机制  

Exploring the Therapeutic Effect and Mechanism of Huanglian Wendan Decoction on Insomnia Rats Based on Neurotransmitter and 5-HT_(1A)/Gα_(i/o)/cAMP Signaling Pathway

作　　者：李亮[1] 王珠强[2] 胡莹[2] 戴卫波[2] 郭文贤 陈巧玲 王家赐[1] LI Liang;WANG Zhuqiang;HU Ying;DAI Weibo;GUO Wenxian;CHEN Qiaoling;WANG Jiaci(Zhongshan Hospital of Traditional Chinese Medicine,Geriatrics and Sleep Specialist,Zhongshan 528401 Guangdong,China;Zhongshan Hospital of Traditional Chinese Medicine,Traditional Chinese Medicine Pharmacology Laboratory,Zhongshan 528401 Guangdong,China)

机构地区：[1]中山市中医院老年病睡眠专科,广东中山528401 [2]中山市中医院中药药理实验室,广东中山528401

出　　处：《中药新药与临床药理》2023年第5期591-598,共8页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：广东省中医药局课题项目(20201379);中山市科技计划项目(2019B1016)。

摘　　要：目的研究黄连温胆汤对失眠大鼠的行为学指标、神经递质水平及5-HT_(1A)/Gα_(i/o)/cAMP信号通路的影响,探讨其对失眠大鼠的治疗作用及机制。方法将SD雄性大鼠随机分为正常组、模型组、地西泮组(2 mg·kg^(-1))及黄连温胆汤低、高剂量组(5.85、11.70 g·kg^(-1))。黄连温胆汤低、高剂量组模型复制前给予相应剂量的黄连温胆汤进行预防性治疗8 d。除正常组外,其余各组腹腔注射对氯苯丙氨酸(DL-4-Chlorophenylalanine,PCPA)复制失眠模型,成模后给予相应剂量药物干预治疗8 d。观察大鼠每日状态,监测体质量,进行旷场实验(行为学监测)并称取全脑质量计算全脑系数。ELISA法检测血清中脑源性神经营养因子(BDNF)及神经胶质纤维酸性蛋白(GFAP)含量,脑干中去甲肾上腺素(NE)、多巴胺(DA)、γ氨基丁酸(GABA)及谷氨酸(Glu)含量,下丘脑中5-羟色胺(5-HT)、5-羟基吲哚乙酸(5-HIAA)及环一磷酸腺苷(cAMP)含量。Western Blot法检测下丘脑中5-HT_(1A)受体蛋白和Gα(i)蛋白表达水平。结果①与正常组比较,模型组大鼠活动总距离、总时间均明显增加(P<0.01),中心区域持续时间明显减少(P<0.01);全脑系数明显降低(P<0.01);血清中BDNF含量明显降低(P<0.01),GFAP含量明显升高(P<0.01);脑干中NE、DA、Glu及Glu/GABA水平均明显升高(P<0.01),GABA水平明显降低(P<0.01);下丘脑中5-HT、5-HIAA及cAMP水平均明显降低(P<0.01),5-HT_(1A)受体蛋白表达水平明显下调(P<0.01),Gα(i)蛋白表达水平明显上调(P<0.01)。②与模型组比较,黄连温胆汤各剂量组大鼠活动总距离、总时间均明显减少(P<0.01),高剂量组大鼠中心区域持续时间明显增加(P<0.01);高剂量组大鼠全脑系数明显升高(P<0.01);各剂量组大鼠血清中BDNF含量明显升高(P<0.01),GFAP含量明显降低(P<0.01);各剂量组大鼠脑干中DA及Glu/GABA水平明显降低(P<0.05,P<0.01),高剂量组的NE及Glu水平明显降低(P<0.01)而GABA水平均明�Objective To study the effect of Huanglian Wendan Decoction(HLWDD)on behavioral indexes,neurotransmitter levels and 5-HT_(1A)/Gα_(i/o)/cAMP signaling pathway in insomnia rats,and to explore the therapeutic effect and mechanism of HLWDD on insomnia rats.Methods SD male rats were randomly divided into normal group,model group,Diazepam group(2 mg·kg^(-1)),low-and high-dose groups of HLWDD(5.85,11.70 g·kg^(-1)).Rats in the low-and high-dose groups of HLWDD were given the corresponding dose of HLWDD for 8 consecutive days before establishing model.Except for the normal group,the rats in the other groups were injected intraperitoneally with DL-4-chlorophenylalanine(PCPA)to establish insomnia model.After the model was copied successfully,all rats were treated with the corresponding dose of drugs for 8 days.Daily state of rats was observed,body mass was monitored,open field test(behavioral monitoring)was conducted,and whole-brain mass was taken to calculate the whole-brain coefficient.ELISA method was used to detect the contents of brain-derived neurotrophic factor(BDNF)and glial fibrillary acidic protein(GFAP)in serum,the contents of norepinephrine(NE),dopamine(DA),gamma aminobutyric acid(GABA)and glutamate(Glu)in brain stem,as well as the contents of 5-hydroxytryptamine(5-HT),5-hydroxyindoleacetic acid(5-HIAA)and cyclic adenosine monophosphate(cAMP)in hypothalamus.The expression levels of 5-HT_(1A) receptor protein and Gα(i)protein in hypothalamus were detected by Western Blot method.Results①Compared with the normal group,total travel distance and total travel time of the rats in the model group were significantly increased(P<0.01),and the duration of the central area was significantly decreased(P<0.01),the whole-brain coefficient was significantly decreased(P<0.01).The content of BDNF in serum was significantly decreased,and the content of GFAP was significantly increased(P<0.01).The levels of NE,DA,Glu and Glu/GABA in the brainstem were significantly increased,and GABA level was significantly decreased(P<0.01).

关 键 词：黄连温胆汤 对氯苯丙氨酸(PCPA) 失眠 神经递质 5-HT_(1A)/Gα_(i/o)/cAMP信号通路 大鼠 

分 类 号：R285.5[医药卫生—中药学]