摘要
目的 本实验通过行为学、转录组学和肠道菌组学,探讨廖氏化风丹抗小鼠焦虑的作用及机制,为其用于抗焦虑提供实验依据。方法 3月龄雄性昆明小鼠给予普通饲料、古方(0.5 g/kg)、现代方(0.5 g/kg)、现代减量方(0.25 g/kg)、未发酵方(0.5 g/kg)饲料,灌胃给予阳性药地西泮各7 d。采用旷场和高架十字迷宫检测行为学,提取大脑RNA进行转录组学测序,提取结肠内容物DNA进行16S rRNA测序,并通过RT-qPCR验证。结果 行为学结果显示化风丹古方、现代方抗焦虑作用显著,而现代减量方和未发酵方抗焦虑作用弱。转录组学结果显示各配方化风丹均产生与地西泮类似调节基因表达的作用,PCA分析显示与对照组相比,各配方化风丹的基因表达谱明显不同,与地西泮类似。与对照组相比,给药组约有200 DEGs,化风丹组以下调偏多。用RT-qPCR验证8个选择基因,化风丹古方、现代方均产生与地西泮类似的基因调节作用,而现代减量方和未发酵方调节作用下降。16S rRNA测序结果显示地西泮升高Simpson,下调Chao和Shannon,但化风丹组变化不明显。RT-qPCR选择验证4个肠菌,结果显示地西泮增加ASV203,但下调ASV230,ASV112和ASV154。化风丹古方、现代方和未发酵方均产生与地西泮类似的调节作用,而现代减量方没有变化。结论 化风丹古方和现代方均能产生和地西泮类似的抗焦虑作用,其机制与调节脑基因表达和肠道菌群有关。未发酵方作用减弱,而现代减量方无效。
Objective Hua-Feng-Dan is a Chinese medicine in the treatment of epilepsy and neurodegeneration.Our prior studies demonstrated its protection against Pentylenetetrazol kindling mice,similar to anxiolytic drug Diazepam.This study aimed to examine the anxiolytic effects of Hua-Feng-Dan recipes in comparison to Diazepam,and used RNA-seq and 16S rRNA-seq to explore the molecular mechanisms.Methods Mice were given Original,Reduced,and Nonfermented Hua-Feng-Dan via feed at the clinical dose(0.5 g/kg for 7 days)and Diazepam was used as positive control.The Open Field test and Elevated Plus Maze were used to access anxiolytic effects.At the end of experiments,the brain and colon content were collected for RNA-seq and 16S rRNA-seq,followed by bioinformatics and qPCR verification.Results Hua-Feng-Dan recipes were effective in producing anxiolytic effects,with original the best and half of reduced(0.25 g/kg)ineffective.PCA revealed brain gene expression patterns were distinct from Control,and approximate 200 DEGs under FDR<0.01 were evident after treatments.qPCR verified 8 of selected genes,the original and reduced Hua-Feng-Dan produced similar effects as Diazepam,while nonfermented and half of reduced were less effective.In gut microbiota analysis,Diazepam increased Simpson,tended to decrease Chao and Shannon indexes,while Hua-Feng-Dan recipes were ineffective.qPCR verified 4 selected microbiome,Diazepam increased ASV203,decreased ASV230,ASV112 and ASV154.Original,reduced,and nonfermented Hua-Feng-Dan produced similar effects,while half of reduced was ineffective.Conclusion Hua-Feng-Dan recipes were effective in producing anxiolytic effects in normal mice,with brain gene expression and gut microbiota modulation in a fashion similar to Diazepam,which could be one of mechanisms for Hua-Feng-Dan to exert anti-epilepsy effects.
作者
李文轲
刘波
赵伟
张蒙
徐尚福
刘杰
李晋
Li Wenke;Liu Bo;Zhao Wei;Zhang Meng;Xu Shangfu;Liu Jie;Li Jin(Key Laboratory of Infectious Diseases and Biosafety,Guizhou Provincial Department of Education,Zunyi Medical University,Zunyi Guizhou 563006,China;Key Laboratory for Basic Pharmacology of Ministry of Education,Zunyi Medical University,Zunyi Guizhou 563006,China)
出处
《合肥医科大学学报》
2026年第1期49-57,共9页
Journal of Zunyi Medical University
基金
贵州省科技计划项目[NO:黔科合基础-ZK(2022)一般596]。
