粪菌移植对大鼠脓毒症相关性脑病的影响及机制  

Impact and mechanisms of fecal microbiota transplantation on sepsis associated encephalopathy via intestinal microbiota

作　　者：李素彦[1] 张立涛[2] 徐鑫[2] 李旭蕊 李灿灿 李宁[1] 郭慧 张亚玉 马志超 贾阳娟 许铁岭 李明泉[2] 李建国[2] Li Suyan;Zhang Litao;Xu Xin;Li Xurui;Li Cancan;Li Ning;Guo Hui;Zhang Yayu;Ma Zhichao;Jia Yangjuan;Xu Tieling;Li Mingquan;Li Jianguo(Department of General Medicine,Hebei General Hospital,Hebei 050057,Shijiazhuang,China;Department of Emergency,Hebei General Hospital,Hebei 050057,Shijiazhuang,China)

机构地区：[1]河北省人民医院全科医疗科,河北石家庄050057 [2]河北省人民医院急诊科,河北石家庄050057

出　　处：《中国中西医结合急救杂志》2019年第3期278-283,共6页Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care

基　　金：河北省科技计划项目(17277720D);河北省医学科学研究重点课题计划(20170262);河北省中医药类科研计划(2017060).

摘　　要：目的观察粪菌移植对脓毒症相关性脑病(SAE)大鼠肠道菌群及脑功能的影响.方法选择60只成年雄性SD大鼠,按随机数字表法分为假手术组、模型组、粪菌移植组,每组20只.采用尾静脉注射脂多糖(LPS)10mg/kg的方法复制脓毒症大鼠模型,粪菌移植组通过灌胃移植正常大鼠粪菌.于制模后6d采集大鼠粪便检测肠道菌群水平;并监测脑功能变化,计算脑电图各波形比例;采用蛋白质免疫印迹试验(WesternBlot)和免疫组化法检测各组大鼠脑皮质Iba-1的蛋白和阳性细胞表达.结果①肠道菌群:模型组大鼠肠道菌群α多样性指数物种指数、Chao1指数较假手术组明显降低(物种指数:282±40比473±37,Chao1指数:730±21比837±27,均P<0.05);粪菌移植组较模型组升高(物种指数:461±20比282±40,Chao1指数:840±16比730±21,P<0.05).门、科和属水平分析显示,模型组大鼠肠道菌群中厚壁菌门和梭杆菌属比例较假手术组显著降低〔厚壁菌门:(22.12±1.34)%比(78.01±1.23)%,梭杆菌属:(2.03±0.17)%比(5.03±0.19)%,均P<0.05〕,变形杆菌门、拟杆菌门和氨基酸球菌科、梭杆菌科、肠杆菌科及另枝菌属比例则明显升高〔变形杆菌门:(70.21±2.35)%比(19.45±2.17)%,拟杆菌门:(4.12±0.19)%比(2.50±0.64)%;氨基酸球菌科:(12.51±0.87)%比(1.01±0.12)%,梭杆菌科:(13.62±1.27)%比(2.31±0.19)%,肠杆菌科:(18.24±2.13)%比(4.15±1.51)%;另枝菌属:(4.53±0.27)%比(1.47±0.33)%,均P<0.05〕;而粪菌移植组厚壁菌门和梭杆菌属水平比例则显著高于模型组〔厚壁菌门:(72.14±2.31)%比(22.12±1.34)%,梭杆菌属:(5.01±0.27)%比(2.03±0.17)%,均P<0.05〕,变形杆菌门、拟杆菌门和氨基酸球菌科、梭杆菌科、肠杆菌科比例则明显低于模型组〔变形杆菌门:(14.23±1.98)%比(70.21±2.35)%,拟杆菌门:(3.15±0.18)%比(4.12±0.19)%;氨基酸球菌科:(0.91±0.11)%比(12.51±0.87)%,梭杆菌科:(1.25±0.15)%比(13.62±1.27)%,肠杆菌科:(3.50±0.21)%比(18.24±2.13)%,均P<Objective To observe the effects of fecal microbiota transplantation on intestinal microbiota and brain function in sepsis rats. Methods Sixty male Sprague Dawley (SD) rats were divided into sham operation group, model group and fecal microbiota transplantation (FMT) group by random number table, each group 20 rats. The rat model of sepsis was established by injection of lipopolysaccharide (LPS) 10 mg/kg in tail vein. FMT group received nasogastric infusion of feces from healthy donor. Fecal samples were collected on the 6th day after the modeling to detect the levels of intestinal microbiota composition;the brain function was also evaluated by electroencephalogram (EEG), and the proportion of each waveform in EEG was calculated. After sacrifice of rats in different groups, the brain tissues were taken, the levels of protein expression and positive cells of Iba-1 in brain tissue were detected by Western Blot and immunohistochemistry method. Results ① Intestinal flora analysis showed that: the diversity index and Chaol index of the intestinal microbiota in model group were significantly lower than that in sham operation group (observed species:282±40 vs. 473±37, Chao1 index: 730±21 vs. 837±27, both P < 0.05);compared with the model group, the diversity index and Chaol index in FMT group were obviously higher (observed species: 461±20 vs. 282±40, Chao1 index:840±16 vs. 730±21, both P < 0.05). At phylum, family, genus level analysis showed that the proportion of Firmicutes phylum and Fusobacterium were obviously lower than those of sham operation group [Firmicutes phylum (22.12±1.34)% vs.(78.01±1.23)%, Fusobacterium:(2.03±0.17)% vs.(5.03±0.19)%, both P < 0.05], and the proportions of Proteobacteria, Bacteroidetes phyla and Acidaminococcaceae, Fusobacteriaceae, Enterbacteriacecae, Alistipes were markedly higher in model group [Proteobacteria:(70.21±2.35)% vs.(19.45±2.17)%, Bacteroidetes phyla:(4.12±0.19)% vs.(2.50±0.64)%, Acidaminococcaceae:(12.51±0.87)% vs.(1.01±0.12)%, Fusobacteriaceae:(13.62±1

关 键 词：粪菌移植 肠道菌群 脓毒症 脓毒症相关性脑病 

分 类 号：R285.5[医药卫生—中药学]