古方半夏秫米汤对正常小鼠最大给药量及其对失眠模型大鼠作用的实验研究  

Maximum Dose of Banxia Shumi Decoction(半夏林米汤)on Normal Mice and Its Effect on Insomnia Model Rats

作　　者：刘玉凤 黄枚[2] 南丽红[2] 许晓丽 谢玉玲 郑立升 LIU Yufeng;HUANG Mei;NAN Lihong;XU Xiaoli;XIE Yuling;ZHENG Lisheng(Hospital of Traditional Chinese Medicine of Fuzhou City,Fuzhou 350001;College of Pharmacy,Fujian University of Traditional Chinese Medicine,Fuzhou 350122;The Fifth Hospital of Xiamen,Xiamen 361101)

机构地区：[1]福州市中医院,福州350001 [2]福建中医药大学药学院,福州350122 [3]厦门市第五医院,厦门361101

出　　处：《中药药理与临床》2023年第8期12-18,共7页Pharmacology and Clinics of Chinese Materia Medica

基　　金：福州市科技计划项目(编号:2020-WS-78);福建省卫生健康委员会青年科研课题(编号:2020QNA075);福州市中医院院内科研项目(编号:2020-Ynky-02);福州市卫生健康科技创新平台-福州市中医药传承创新平台(编号:2021-S-wp3)。

摘　　要：目的:观察半夏秫米汤对失眠模型大鼠的治疗作用及其对生物钟基因和神经递质的影响,初步探究其药效物质基础及安全性,阐释临床使用大剂量半夏秫米汤治疗失眠的理论基础,有利于指导临床合理及安全用药。方法:测定半夏秫米汤最大给药量并利用广泛靶向代谢组技术确定其化学成分;采用氯苯丙氨酸(PCPA)诱导大鼠失眠模型,观察半夏秫米汤对失眠模型大鼠的治疗作用及下丘脑组织病理学变化的影响;采用ELISA法检测大鼠下丘脑组织5-羟色胺(5-HT)、多巴胺(DA)、褪黑色素(MT)含量;Western blot法及qRT-PCR法检测大鼠下丘脑组织昼夜运动输出周期(Clock)、脑和肌肉芳烃受体核转录因子样蛋白1(BMAL1)蛋白及mRNA的表达。结果:半夏秫米汤小鼠最大给药量为810 g/kg。鉴定出999种半夏秫米汤代谢物,主要代谢物为氨基酸及其衍生物类(49.81%)、核苷酸及其衍生物类(16.62%)。与正常对照组相比,模型对照组大鼠体质量增加较少,睡眠潜伏期明显延长,睡眠持续时间显著缩短(P<0.01);同时神经元染色变深,细胞大小不一,形状变长,核皱缩;下丘脑5-HT、MT的含量明显降低,DA含量明显升高,Clock、BMAL1表达明显上调(P<0.05或P<0.01);与模型对照组比较,半夏秫米汤8.1、16.2、32.4 g/kg组大鼠体质量增加较大,睡眠潜伏期缩短和睡眠持续时间明显延长(P<0.05或P<0.01),神经元损伤改善;半夏秫米汤16.2、32.4 g/kg组MT的含量明显升高,而DA的含量明显降低(P<0.05或P<0.01);半夏秫米汤32.4 g/kg组5-HT的含量显著升高(P<0.01);半夏秫米汤8.1、16.2、32.4 g/kg组Clock、BMAL1蛋白及mRNA表达明显下调(P<0.05或P<0.01)。结论:半夏秫米汤安全范围较大,其代谢产生的氨基酸类和核苷酸类代谢产物可能是其药理作用的重要物质基础。半夏秫米汤具有较好的抗失眠作用,该作用可能与其降低大鼠下丘脑组织中DA含量,增加5-HT、MT含量,抑制下丘脑ClObjective:To observe the therapeutic effect of Banxia Shumi Decoction(半夏米汤)on a rat model of insomnia,investigate its influence on circadian clock genes and neurotransmiters,and explore its pharmacological basis and safety,thereby explaining the theoretical foundation of clinical use of the maximum dose of Banxia Shumi Decoction for the treatment of insomnia and providing insights for the clinical rational and safe medication.Methods:The maximum dose of Banxia Shumi Decoction was determined,and its chemical composition was identified using comprehensive targeted metabolomics.Insomnia was induced in rats using para-chlorophenylalanine(PCPA),and the therapeutic effect of Banxia Shumi Decoction on insomnia and its impact on hypothalamic tissue pathology were observed.The levels of serotonin(5-HT),dopamine(DA),and melatonin(MT)in the hypothalamic tissues of rats were measured using ELISA.The protein and mRNA expression of circadian locomotor output cycles kaput(Clock)and brain and muscle aryl hydrocarbon receptor nuclear translocator-like protein 1(Bmall)in the hypothalamic tissues was assessed using Western blot and qRT-PCR,respectively.Results:The maximum dose of Banxia Shumi Decoction in mice was 810 g/kg.A total of 999 metabolites of Banxia Shumi Decoction were identified,with the main metabolites belonging to the amino acids and derivatives(49.81%)and nucleotides and derivatives(16.62%).Compared with the rats in the normal control group,those in the model control group showed less body gain,significantly prolonged sleep latency,and significantly reduced sleep duration(P<0.01).Additionally,neuronal staining became darker,cell sizes varied,shapes elongated,and nuclei appeared wrinkled.The levels of 5-HT and MT in the hypothalamus were significantly decreased,while the level of DA was significantly increased.The expression of Clock and Bmall was significantly upregulated(P<0.05 or P<0.01).Compared with rats in the model control group,those treated with Banxia Shumi Decoction at 8.1 g/kg,16.2 g/kg,and 32.4 g

关 键 词：半夏秫米汤 失眠 最大给药量 褪黑色素 昼夜运动输出周期 脑和肌肉芳烃受体核转录因子样蛋白1 

分 类 号：R285.5[医药卫生—中药学]