摘要
目的运用大鼠粪便16s rRNA测序和非靶向代谢组学分析脓毒症大鼠肠道微生物组成及代谢产物,初步探究肠道菌群及其代谢物对脓毒症炎症反应和多器官损伤的影响及潜在机制.方法将10只健康雄性Wistar大鼠按随机数字表法分为假手术组(Sham组)和脓毒症模型组(CLP组),每组5只.通过盲肠结扎穿孔术(CLP)建立脓毒症大鼠模型.制模后24h处死动物,取外周血采用酶联免疫吸附试验(ELISA)检测血清炎症因子肿瘤坏死因子-α(TNF-α)和白细胞介素-6(IL-6)的含量;采用苏木素-伊红(HE)染色观察肺、肾组织病理改变并进行病理评分;取粪便样本,采用16s rRNA高通量测序和非靶代谢组学筛选可能在疾病结局中发挥重要作用的微生物群、代谢物和信号通路;Spearman相关分析对肠道菌群与非靶向代谢进行联合分析.结果与Sham组相比,CLP组肺组织和肾脏组织病理损伤程度明显加重[肺组织评分(分):3.60±0.80比0.00±0.00,肾组织评分(分):2.40±0.80比0.00±0.00,均P<0.01];外周血中TNF-α和IL-6含量明显升高[TNF-α(ng/L):248.12±55.98比143.28±36.57,IL-6(ng/L):260.26±39.47比116.01±26.43,均P<0.05];肠道菌群物种多样性显著降低;摩根菌属、拟杆菌属和大肠埃希菌-志贺菌属的相对丰度显著增加,毛螺菌NK4A136属、瘤胃球菌属、罗姆布茨菌属和罗氏菌属的相对丰度显著降低;肠道微生物群苯丙氨酸、酪氨酸和色氨酸生物合成及胆汁分泌显著增加,而初级胆汁酸生物合成显著降低;差异代谢物与差异菌群之间存在显著相关性.结论脓毒症可以造成大鼠肠道菌群和粪便代谢物特征发生明显变化,该结果为寻求治疗脓毒症的新靶点提供了转化研究基础.
Objective To analyze the composition and metabolites of gut microbiota in septic rats by fecal 16s rRNA sequencing and untargeted metabolomics,and to preliminarily explore the effect and potential mechanism of gut microbiota and its metabolites on inflammatory response and multiple organ damage in sepsis.Methods Ten males healthy male Wistar rats were randomly divided into a sham operated group(Sham group)and sepsis model group(CLP group)using a random number table method,with 5 rats in each group.A rat sepsis model was established by cecal ligation and perforation(CLP)method.The animals were sacrificed 24 hours after modeling,the levels of tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6)in peripheral blood were detected by enzyme-linked immunosorbent assay(ELISA).Hematoxylin-eosin(HE)staining was used to observe the pathological changes of lung and kidney tissues,and the pathological scores were evaluated.Fecal samples were collected,and 16s rRNA high-throughput sequencing and non-targeted metabolomics were used to screen microbiota,metabolites and potential signal pathways that may play an important role in disease outcomes.Spearman correlation analysis was conducted to jointly analyze the gut microbiota and non-targeted metabolism.Results Compared with the Sham group,the degree of pathological damage to lung and kidney tissues in the CLP group was significantly increased(lung tissue score:3.60±0.80 vs.0.00±0.00,kidney tissue score:2.40±0.80 vs.0.00±0.00,both P<0.01),the level of IL-6 and TNF-αin peripheral blood significantly increased[TNF-α(ng/L):248.12±55.98 vs.143.28±36.57,IL-6(ng/L):260.26±39.47 vs.116.01±26.43,both P<0.05],the species diversity of intestinal flora of rats in the CLP group was significantly reduced,the relative abundance of Morganella,Bacteroides and Escherichia-Shigella were significantly increased,and the relative abundance of Lachnospiraceae NK4A136,Ruminococcus,Romboutsia and Roseburia were significantly reduced.In addition,the biosynthesis and bile secretion of phenyl
作者
张萍
杨磊
卓玉珍
崔凌志
高巧营
周旻
戈立秀
李家瑞
Zhang Ping;Yang Lei;Zhuo Yuzhen;Cui Lingzhi;Gao Qiaoying;Zhou Min;Ge Lixiu;Li Jiarui(Department of Nephrology,Tianjin Nankai Hospital,Tianjin 300100,China;Integrated Traditional Chinese and Western Medicine Acute Abdominal Disease Research Institute,Tianjin Nankai Hospital,Tianjin 300100,China;Graduate School of Tianjin Medical University,Tianjin 300070,China;Department of Clinical Laboratory,Tianjin Nankai Hospital,Tianjin 300100,China)
出处
《中华危重病急救医学》
CAS
CSCD
北大核心
2023年第9期927-932,共6页
Chinese Critical Care Medicine
基金
天津市自然科学基金(18JCQNJC13400)
天津市卫建委中西医结合课题(2021093,2021044)。
