基于“微生物群-肠-脑轴”探讨复方抗焦虑胶囊对慢性束缚应激大鼠的抗焦虑作用及机制  

Fufang Kangjiaolv Capsules Treat Anxiety in Rat Model of Chronic Restraint Stress via Microbiota-gut-brain Axis

作　　者：范文昕 姜婷月 王宇 张鸽 芦艺凡 刘朦朦 李佳园 马仁芝 石晋丽[1] FAN Wenxin;JIANG Tingyue;WANG Yu;ZHANG Ge;LU Yifan;LIU Mengmeng;LI Jiayuan;MA Renzhi;SHI Jinli(School of Chinese Materia Medica,Beijing University of Chinese Medicine,Beijing 102488,China)

机构地区：[1]北京中医药大学中药学院,北京102488

出　　处：《中国实验方剂学杂志》2025年第4期95-107,共13页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金面上项目(82073971)。

摘　　要：目的:观察复方抗焦虑胶囊对慢性束缚应激(CRS)模型大鼠焦虑样行为的干预作用,探究复方抗焦虑胶囊通过“微生物群-肠-脑轴”发挥抗焦虑的作用机制。方法:采用CRS建立大鼠焦虑模型,将大鼠分为正常组、模型组、阳性药组(地西泮,1 mg·kg^(-1))及复方抗焦虑胶囊低、中、高剂量组(0.75、1.5、3 g·kg^(-1))。给药14 d后进行高架十字迷宫实验(EPM)、旷场实验(OFT)、明暗箱实验(LDB)和埋珠实验(MBT)。采用苏木素-伊红(HE)染色观察大鼠海马及结肠病理变化,尼氏(Nissl)染色观察海马神经元损伤情况;16S rRNA测序技术检测各组大鼠肠道菌群多样性变化;实时荧光定量聚合酶链式反应(Real-time PCR)检测大鼠结肠中功能蛋白闭锁小带蛋白-1(ZO-1)及闭合蛋白(Occludin)mRNA的表达;酶联免疫吸附测定法(ELISA)检测大鼠结肠、血清及海马体中炎症因子肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)、IL-1β的含量;蛋白免疫印迹法(Western blot)检测大鼠结肠组织ZO-1、Occludin、核转录因子-κB p65(NF-κB p65)及大鼠海马组织NF-κB p65和脑源性神经营养因子(BDNF)蛋白表达。结果:与正常组比较,模型组大鼠进入高架十字迷宫时间及次数比例,进入旷场中央区域的时间及次数,进入明箱时间及穿越明暗箱次数及未被掩埋珠子的个数明显降低(P<0.05,P<0.01);与模型组比较,复方抗焦虑胶囊给药组能在不同程度上改善CRS模型大鼠的焦虑状态,其中高剂量组效果最优,能明显增加大鼠在高架十字迷宫中进入开臂的时间及次数比例(P<0.05),提升大鼠在旷场中进入中央区域的次数(P<0.05),显著增加其进入明箱时间、穿越明暗箱次数和未被掩埋珠子的个数(P<0.01),并能缓解大鼠海马神经元及结肠病理损伤。16S rRNA结果表明,模型组厚壁菌门、Deferribacterota、罗姆布茨菌属(Romboutsia)、Phascolarctobacterium占比升高,拟杆菌门(Bavcteroidota)、乳杆菌�Objective:To observe the intervention effect of Fufang Kangjiaolv capsules on anxiety-like behaviors in the rat model of chronic restraint stress(CRS)and explore the mechanism underlying the anti-anxiety effect via the microbiota-gut-brain axis.Methods:Rats were assigned into blank,model,positive drug(diazepam,1 mg·kg^(-1)),and low-,medium-,and high-dose(0.75,1.5,3 g·kg^(-1),respectively)Fufang Kangjiaolv capsules groups.After 14 days of administration,the elevated plus maze test,open field test,light and dark box test,and marble burying test were performed.Hematoxylin-eosin staining was employed to observe the pathological changes in the hippocampus and colon of rats,and Nissl staining was conducted to observe the damage of hippocampal neurons.The gut microbiota was analyzed by 16S rRNA gene sequencing.Real-time fluorescence quantitative polymerase chain reaction(Real-time PCR)was employed to determine the mRNA levels of zonula occludens-1(ZO-1)and occludin in the colon of rats.The levels of tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),and interleukin-1β(IL-1β)in the colon,serum,and hippocampus were determined by enzyme-linked immunosorbent assay.Western blot was employed to determine the protein levels of ZO-1,occludin,nuclear factor-κB p65(NF-κB p65)in the colon tissue and NF-κB p65 and brainderived neurotrophic factor(BDNF)in the hippocampal tissue.Results:Compared with the blank group,the model group showed reductions in the time and frequency ratio of rats entering the elevated plus maze,the time and frequency of rats entering the central area of the open field,the time of entering the open box,the times of passing through the light and dark box,and the number of unburied beads(P<0.05,P<0.01).Compared with the model group,Fufang Kangjiaolv capsules ameliorated the anxiety of the model rats to varying degrees,and the high-dose group had the best effect,with increases in the proportions of time and frequency of rats entering the open arm in the elevated plus maze(P<0.05),the number of rats ente

关 键 词：复方抗焦虑胶囊 焦虑症 慢性束缚应激模型 肠道菌群 肠道屏障 炎症 

分 类 号：R285[医药卫生—中药学] R289[医药卫生—中医学] R259