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聂慧芳, 彭株丽, 游建凯, 杨彤, 吴海辉, 赖嘉豪, 葛金文, 梅志刚. 缺血性脑卒中肠道微生物移植致菌群失调大鼠模型建立[J]. 中国公共卫生, 2022, 38(3): 314-319. DOI: 10.11847/zgggws1133778
引用本文: 聂慧芳, 彭株丽, 游建凯, 杨彤, 吴海辉, 赖嘉豪, 葛金文, 梅志刚. 缺血性脑卒中肠道微生物移植致菌群失调大鼠模型建立[J]. 中国公共卫生, 2022, 38(3): 314-319. DOI: 10.11847/zgggws1133778
NIE Hui-fang, PENG Zhu-li, YOU Jian-kai, . Establishment of a microbial transplantation-induced dysbacteriosis model in rats with ischemic stroke[J]. Chinese Journal of Public Health, 2022, 38(3): 314-319. DOI: 10.11847/zgggws1133778
Citation: NIE Hui-fang, PENG Zhu-li, YOU Jian-kai, . Establishment of a microbial transplantation-induced dysbacteriosis model in rats with ischemic stroke[J]. Chinese Journal of Public Health, 2022, 38(3): 314-319. DOI: 10.11847/zgggws1133778

缺血性脑卒中肠道微生物移植致菌群失调大鼠模型建立

Establishment of a microbial transplantation-induced dysbacteriosis model in rats with ischemic stroke

  • 摘要:
      目的   建立一种肠道菌群失调大鼠模型,为脑缺血损伤肠道菌群失调研究提供方法学支持。
      方法   将SPF级SD大鼠分为正常组、供体组和受体组,对供体组大鼠予以大脑中动脉栓塞术(MCAO),术后72 h取盲肠菌群作为移植物;对受体组大鼠灌胃硫酸链霉素消耗肠道微生物后,将供体组的盲肠菌群灌胃移植给受体组,1次/d,共3 d。取3组大鼠盲肠内容物进行革兰染色镜检;取正常组、受体组大鼠回肠、盲肠、横结肠内容物及供体组大鼠盲肠内容物进行16S rDNA高通量测序,检测其微生物结构及特性,并进行生物信息学分析。
      结果   正常组大鼠盲肠、横结肠菌群的物种多样性相对回肠显著增加(P < 0.01),3个肠段中的菌群均主要由厚壁菌门组成。与正常组比较,供体组大鼠盲肠菌群的物种多样性显著减少,门水平上物种组成分析显示,厚壁菌门显著减少、变形菌门显著增加(P < 0.01)。与正常组比较,受体组大鼠盲肠及横结肠菌群的物种多样性显著减少(P < 0.01);受体组大鼠盲肠菌群与横结肠菌群结构相近,主要由拟杆菌门组成;受体组大鼠回肠、盲肠、横结肠菌群与供体组大鼠盲肠菌群在α多样性ACE指数、β多样性均无统计学差异(P > 0.05),两组大鼠的菌群结构相近。
      结论   MCAO 72 h大鼠盲肠菌群灌胃移植可建立一种与脑缺血微生物结构及特性类似的肠道菌群失调大鼠模型。

     

    Abstract:
      Objective  To establish a rat model of intestinal dysbacteriosis and provide methodological support for the study of intestinal dysbacteriosis after cerebral ischemia.
      Methods  Totally 19 male specific pathogen free (SPF) Sprague-Dawley (SD) rats were divided into a normal group (n = 5), a donor group (n = 9) and a recipient group (n = 5). The rats of donor group were subsequently assigned into 3 groups and middle cerebral artery occlusion (MCAO) was performed in the rats on the first, second and third day after a 3-day adaptive feeding and cecal flora were taken as grafts 72 hours after the MCAO operation. After gavage administration of streptomycin sulfate for exhausting intestinal microbes, the rats of the recipient group were administered with the grafts by gavage once a day continually for three days. The cecal specimens of all the rats of the three groups were examined with gram staining microscopy. The contents of ileum, cecum and transverse colon of normal and recipient rats and cecum contents of donor rats were sampled for 16S rDNA high-throughput sequencing and the microbial structure and characteristics of the contents were detected and analyzed with bioinformatics.
      Results  Compared with that of the ileum, the species diversity of cecum and transverse colon flora in normal rats increased significantly (P < 0.01) and the flora in the three intestinal segments were mainly composed of thick-walled bacteria. Compared with that of the normal rats, the species diversity of cecal flora in the donor rats decreased significantly; the species composition analysis at phylum level showed that the Firmicutes decreased significantly and the Proteobacteria increased significantly (both P < 0.01). Compared with that of the normal rats, the species diversity of cecum and transverse colon flora in the recipient rats decreased significantly (P < 0.01). The cecal flora of the recipient rats was similar to that of the transverse colon, mainly composed of Bacteroides. There were no significant differences in α diversity ACE index and β diversity among the ileum, cecum and transverse colon flora of the recipient rats and the cecum flora of the donor rats (P > 0.05 for all), but the flora structure of the rats of the two groups were similar.
      Conclusion   Gavage administration of cecal flora 72 hours after MCAO operation could be used to establish a rat model of intestinal flora imbalance similar to the structure and characteristics of microorganisms of the rats with ischemic stroke.

     

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