产芽孢菌调控五羟色胺促进胃肠动力的机制研究  

Study on the Mechanism of Spore-forming Microbes Promoting Gastrointestinal Motility through Regulating Serotonin Content

作　　者：黄薏佳 陈燕敏 陈丰连[1] 王术玲[1] HUANG Yi-jia;CHEN Yan-min;CHEN Feng-lian;WANG Shu-ling(School of Pharmaceutical Sciences,Guangzhou University of Chinese Medicine,Guangzhou 510006,Guangdong,China)

机构地区：[1]广州中医药大学中药学院,广东广州510006

出　　处：《食品研究与开发》2022年第14期79-87,共9页Food Research and Development

基　　金：国家自然科学基金面上项目(82174257)。

摘　　要：为探究产芽孢菌对胃肠运动的调节作用及其潜在机制,利用氯仿提取法在体外提取健康志愿者粪便中的产芽孢菌菌液,并通过聚合酶链式反应验证产芽孢菌菌液的提取效果。运用抗生素鸡尾酒法建立伪无菌(pseudo-germfree,PGF)小鼠模型,并用粪便菌悬液涂板实验及肠杆菌基因间保守重复序列-聚合酶链式反应(enterobacterial repetitive intergenic consensus sequences-polymerase chain reaction,ERIC-PCR)进行模型评价。PGF小鼠造模第11天,小鼠粪便悬液涂板无明显菌落且ERIC-PCR条带显著减少,表明实验造模成功。成模后,将小鼠随机分为模型组和产芽孢菌定殖组,进行两周的产芽孢菌定殖,采用荧光定量核酸扩增检测方法检测产芽孢菌定殖情况。通过小肠推进率和胃残留率实验测定胃肠动力,并收集小鼠结肠和血浆,检测其中五羟色胺(5-hydroxytryptamine,5-HT)的含量以及结肠中色氨酸羟化酶1(tryptophan hydroxylase 1,TPH1)的表达水平。实验结果表明,产芽孢菌定殖后,小鼠胃残留率显著下降(P<0.05),小肠推进率显著升高(P<0.01),粪便中瘤胃球菌属、扭链瘤胃球菌属、多尔氏菌属含量显著上升,结肠和血浆中5-HT含量明显提高,但是结肠中TPH1表达明显下调。表明产芽孢菌可通过提高肠道和血浆中5-HT的含量水平以促进胃肠运动。The regulation of gastrointestinal motility by spore-forming microbes and the potential mechanism were investigated.Spore-forming microbes were extracted in vitro from the feces of healthy volunteers using the chloroform.The extraction efficiency was confirmed by polymerase chain reaction(PCR).The pseudo germ-free mouse model created using the antibiotic cocktail approach was evaluated by a fecal bacteria suspension coating experiment and Enterobacterial Repetitive Intergenic Consensus Sequences-PCR(ERIC-PCR).The fecal suspension coating plate collected and processed on day 11 of PGF mice modeling did not produce visible colonies.The ERIC-PCR bands were greatly diminished,demonstrating the successful establishment of the model.The mice were randomly divided into model and spore-forming microbe groups.Transplantation in the spore-forming microbes group was performed by gavage.The colonization of spore-forming microbes was confirmed by quantitative PCR.Intestinal thrust and residual gastric rates were used to assess intestinal motility.The contents of 5-hydroxytryptamine(5-HT,serotonin)and tryptophan hydroxylase 1(TPH1)in the colon and plasma were determined.Following colonization,the residual gastric rate of mice was significantly decreased(P<0.05),while the intestinal thrust rate was significantly increased(P<0.01).The contents of Ruminococcus,Ruminococcus_torques_group,and Dorea in feces were significantly increased.The levels of 5-HT in the colon and plasma were significantly increased,but the expression of TPH1 in the colon was significantly downregulated.The results indicate that spore-forming microbes promote gastrointestinal motility by increasing intestinal and plasma serotonin levels.

关 键 词：产芽孢菌 胃肠运动功能 胃残留率 小肠推进率 五羟色胺 色氨酸羟化酶1 

分 类 号：TS201.3[轻工技术与工程—食品科学]