摘要
目的:探讨高温高湿环境下大鼠肠道菌群结构和功能表达,及其与血清脑肠肽指标间的相关性。方法:将16只成年SD雄性大鼠随机分成2组,常温常湿组和高温高湿组,每组8只。常温常湿组放置于温度(25±2)℃、湿度(50±5)%的恒温恒湿箱中,高温高湿组放置于温度(40±2)℃、湿度(70±5)%的恒温恒湿箱中,饲养7 d,期间正常喂食饮水。实验收集两组SD大鼠粪便样品于1.5 mL离心管,并通过腹主动脉采集血液样本。采用16S rDNA和宏基因组测序法对2组大鼠粪便样品进行分析,明确高温高湿环境对菌群结构和功能的影响。采用酶联免疫吸附测定法(ELISA)测定大鼠血清胆囊收缩素(CCK)水平;Spearman相关性分析菌群与脑肠肽间的相关性。结果:动物行为学观察表明高温高湿组大鼠躁动不安,随着时间延长出现大鼠前爪挠面部且精神差的情况,大鼠活动也逐渐减弱。16S rDNA和宏基因组测序结果显示,与常温常湿组相比,高温高湿环境能够显著降低肠道菌群的Alpha和Beta多样性,引起厚壁菌门/拟杆菌门的比值升高。物种差异分析获得核心差异菌群,其中上调菌群为棒状杆菌、另枝菌属,下调菌群为普雷沃氏菌、考拉杆菌、毛螺旋菌、瘤胃球菌;功能分析结果表明高温高湿主要影响精氨酸和脯氨酸代谢、卟啉代谢、视黄醇代谢等新陈代谢过程及生物合成,还涉及糖酵解/糖新生、ABC转运蛋白、核糖体等。ELISA测定结果显示高温高湿组SD大鼠血清CCK含量较常温常湿组显著降低,并与核心差异菌群呈现相关性。结论:高温高湿环境可引起大鼠肠道菌群结构和功能异常,进而通过核心差异菌群影响CCK水平变化,影响大鼠情绪的调控。功能富集分析表明,高温高湿环境下大鼠肠道差异菌与大鼠情绪变化具有相关性,提示与神经性疾病有重要的关系。该研究丰富了环境对肠道菌群影响的认识,同时对临床诊断和研究提供了支撑。
OBJECTIVE:Genomics was used to explore the structure and functional expression of gut microbiota in rats under high temperature and humidity environments,as well as their correlation with serum brain-gut peptide indicators.METHODS:Sixteen adult male SD rats were randomly divided into two groups:the normal temperature and humidity group and the high temperature and humidity group.The normal temperature and humidity group were placed in a constant temperature and humidity box with a temperature of(25±2)℃ and a humidity of(50±5)%,while the high temperature and humidity group with a temperature of(40±2)℃ and a humidity of(70±5)% for 7 days.During this period,the rats were fed and watered normally.The fecal samples of the two groups were collected in a 1.5 mL centrifuge tube,and serum samples were collected through the abdominal aorta.16S rDNA and metagenomic sequencing methods were used to analyze fecal samples of rats in the two groups.Changes in bacterial community structure and function under high temperature and humidity environments were identified.Enzyme linked immunosorbent assay(ELISA)was used to measure serum cholecystokinin(CCK)levels in rats.Spearman correlation analysis was applied to investigate the correlation between differential microbiota and brain gut peptides.RESULTS:Animal behavioral observations indicated that rats in the high temperature and humidity group were restless,with their front paws scratching their faces and poor mental state over time,and their activity was gradually decreased.The 16S rDNA and metagenomic sequencing results showed that the high temperature and humidity environment significantly reduced the alpha and beta diversity of the intestinal microbiota compared with the normal temperature and humidity group,leading to an increase in the ratio of Firmicutes to Bacteroidetes.The core differential microbiota was obtained through species difference analysis,the up-regulated microbiota were Corynebacterium and Streptomyces,and the down-regulated microbiota were Prevotella,Corynebacterium,Helicobacter pilosum,and Ruminococcus.Functional analysis results indicated that high temperature and humidity mainly affect metabolic processes such as arginine and proline metabolism,porphyrin metabolism,retinol metabolism,and biosynthesis,as well as glycolysis/glycogenesis,ABC transporters,ribosomes,etc.The ELISA test results showed that the serum CCK content of SD rats in the high-temperature and high humidity group was significantly reduced.CONCLUSION:High temperature and humidity environments caused abnormalities in the structure and function of microbial communities significantly,which in turn affected the changes in CCK levels through core differential microbial communities,thereby affecting emotional regulation.Functional enrichment analysis showed that there was a correlation between gut microbiota and emotional changes in high temperature and humidity environments,which in turn had an important relationship with neurological diseases.This study enriches the understanding of the impact of the environment on gut microbiota and provides support for clinical diagnosis and research.
作者
朱博
于智颖
杨洁
李思远
贺江华
孙艺璇
亓云鹏
罗闽
董玲
徐文娟
ZHU Bo;YU Zhiying;YANG Jie;LI Siyuan;HE Jianghua;SUN Yixuan;QI Yunpeng;LUO Min;DONG Ling;XU Wenjuan(School of Life Sciences,Beijing University of Chinese Medicine,Beijing 102488;School of Chinese Materia Medica,Beijing University of Chinese Medicine,Beijing 102488,China)
出处
《癌变．畸变．突变》
CAS
2024年第3期224-230,236,共8页
Carcinogenesis,Teratogenesis & Mutagenesis
关键词
高温高湿
肠道菌群
宏基因组
脑肠肽
high temperature and humidity
intestinal microbiota
metagenomic
brain gut peptide
