摘要
目的:基于5-羟色胺系统和肠道菌群研究黄连阿胶汤干预失眠的机制。方法:将55只SPF级SD大鼠随机分为正常组、模型组、黄连阿胶汤低、中、高剂量组(1.925、3.85、7.7 g·kg^(-1))和艾司唑仑组(0.1 mg·kg^(-1)),除正常组,其余5组大鼠被放在窄平台剥夺睡眠,每天剥夺12 h,连续21 d。药物干预14 d后,分别通过戊巴比妥钠睡眠协同实验、旷场实验和糖水偏好实验评估大鼠的睡眠情况、探究行为和抑郁样行为;采用酶联免疫吸附测定法(ELISA)检测5-羟色胺(5-HT)、5-羟吲哚乙酸(5-HIAA)、色氨酸羟化酶(TPH)和单胺氧化酶-A(MAO-A)的含量,实时荧光定量聚合酶链式反应(Real-time PCR)检测5-HT1A受体(5-HT1AR)和5-HT2A受体(5-HT2AR)的mRNA表达;基于16S rRNA测序技术检测各组大鼠肠道菌群的差异,采用冗余分析方法揭示5-HT系统与菌群的关联。结果:与正常组比较,模型组大鼠的睡眠潜伏期延长(P<0.05),睡眠维持时间显著缩短(P<0.01),旷场中央区域活动时间显著减少(P<0.01),糖水偏好率明显降低(P<0.05);5-HT、5-HIAA、TPH和MAO-A的含量显著降低(P<0.01),5-HIAA/5-HT显著下降(P<0.01),5-HT1ARmRNA表达显著下调(P<0.01),5-HT2AR mRNA表达明显上调(P<0.05);厚壁菌门的占比降低,拟杆菌门的占比升高,厚壁菌门与拟杆菌门的比值(F/B)明显减小(P<0.05)。与模型组比较,黄连阿胶汤高剂量组的睡眠潜伏期缩短(P<0.01),睡眠维持时间延长(P<0.01),黄连阿胶汤低剂量组的中央区域活动时间增加(P<0.01),糖水偏好率明显升高(P<0.05);黄连阿胶汤高剂量组的5-HT、5-HIAA、TPH和MAO-A的含量增加(P<0.01)、5-HIAA/5-HT升高(P<0.05),5-HT1AR mRNA上调(P<0.01),5-HT2AR mRNA下调(P<0.05);黄连阿胶汤低剂量组的厚壁菌门占比升高,拟杆菌门占比降低,F/B增大。门水平上,5-HT、5-HIAA、MAO-A与厚壁菌门呈正相关,与拟杆菌门呈负相关;属水平上,5-HT、5-HIAA、TPH、MAO-A与普雷沃氏菌属和乳酸杆菌属呈负�
Objective:To explore the mechanism of Huanglian Ejiaotang in intervening in insomnia based on 5-hydroxytryptamine(5-HT)system and gut microbiota.Method:Fifty-five SPF-grade SD rats were randomly divided into normal group,model group,low-,medium-,and high-dose Huanglian Ejiaotang groups(1.925,3.85,and 7.7 g·kg^(-1)),and Estazolam group(0.1 mg·kg^(-1)).Except for those in the normal group,the rats in the other five groups were subjected to sleep deprivation on a narrow platform for 12 hours daily for 21 consecutive days.After 14 days of drug intervention,the sleep,exploratory behavior,and depressive-like behavior of the rats were assessed using the pentobarbital sodium sleep synergistic test,the open field test,and the sugar preference test,respectively.Enzyme-linked immunosorbent assay(ELISA)was used to detect the levels of 5-HT,5-hydroxyindoleacetic acid(5-HIAA),tryptophan hydroxylase(TPH),and monoamine oxidase-A(MAO-A).Real-time polymerase chain reaction(Real-time PCR)was used to measure the mRNA expression of the 5-HT1A receptor(5-HT1AR)and 5-HT2A receptor(5-HT2AR).Differences in gut microbiota among the groups were assessed using 16S rRNA sequencing,and the correlation between the 5-HT system and microbiota was revealed using redundancy analysis.Result:Compared with the normal group,the model group showed a prolonged sleep latency(P<0.05),reduced sleep maintenance(P<0.01),decreased central area activity time in the open field(P<0.01),and reduced sugar preference rate(P<0.05).Moreover,the model group also showed decreased levels of 5-HT,5-HIAA,TPH,and MAO-A(P<0.01),decreased 5-HIAA/5-HT ratio(P<0.01),downregulated mRNA expression of 5-HT1AR(P<0.01),and upregulated mRNA expression of 5-HT2AR(P<0.05).The proportion of Firmicutes decreased,while that of Bacteroidetes increased,leading to a decreased Firmicutes/Bacteroidetes(F/B)ratio(P<0.05).Compared with the model group,the high-dose Huanglian Ejiaotang group exhibited a shortened sleep latency(P<0.01),and increased sleep maintenance(P<0.01).The low-dose Huangl
作者
刁华琼
魏丹
丁海月
张婧
陈雨菲
王敏
朱庆生
李小黎
DIAO Huaqiong;WEI Dan;DING Haiyue;ZHANG Jing;CHEN Yufei;WANG Min;ZHU Qingsheng;LI Xiaoli(Beijing University of Chinese Medicine Third Affiliated Hospital,Beijing 100029,China;Third Hospital of Beijing Chaoyang District,Beijing 100121,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2023年第21期49-58,共10页
Chinese Journal of Experimental Traditional Medical Formulae
基金
第五批全国中医临床优秀人才研修项目(国中医药人教函[2022]1号)
朝阳区科技计划项目(CYSF2205)。
关键词
黄连阿胶汤
睡眠剥夺
5-羟色胺
肠道菌群
动物实验
Huanglian Ejiaotang
sleep deprivation
5-hydroxytryptamine
gut microbiota
animal experimen
