李映荷;孟祥辰;阎鹏光;杨红;李骥;钱家鸣;李景南;
LI Ying-he;MENG Xiang-chen;YAN Peng-guang;YANG Hong;LI Ji;QIAN Jia-ming;LI Jing-nan;Department of Gastroenterology,Peking Union Medical College Hospital,Chinese Academy of Medical Sciences & Peking Union Medical College;Laboratory of Intestinal Microecology and Clinical Translational Research,Chinese Academy of Medical Sciences;

• 1:中国医学科学院北京协和医学院北京协和医院消化内科
• 2:中国医学科学院肠道微生态与临床转化研究重点实验室

摘要(Abstract)：

目的分析接受及不接受糖皮质激素治疗的中重度溃疡性结肠炎(ulcerative colitis,UC)患者肠道菌群的差异,并探讨肠道菌群对静脉糖皮质激素疗效的预测价值。方法回顾性分析2016年11月1日至2018年6月30日期间,在北京协和医院消化内科门诊就诊或住院治疗的中重度UC患者临床资料。根据采集粪便样本时患者是否正在使用糖皮质激素,将患者分为激素组和无激素组。无激素组患者入院后接受静脉激素、口服激素或免疫抑制剂治疗,根据3d后的疗效,将入院后接受足量静脉激素治疗的患者分为治疗有效组和无效组。采用16S rRNA扩增子测序方法对粪便菌群测序分析,采用香农指数评估肠道菌群α多样性,采用Metastats分析比较不同组患者肠道菌群的物种构成差异。结果共35例符合入选和排除标准的中重度UC患者入选本研究,无激素组20例,激素组15例。无激素组中13例患者在采集粪便样本后接受静脉足量糖皮质激素治疗,其中有效组8例,无效组5例。激素组和无激素组患者的肠道菌群α多样性(香农指数:无激素组3. 57±0. 73,激素组3. 03±1. 15,P=0. 123)及物种组成无统计学差异。激素治疗有效组和无效组患者的肠道菌群α多样性亦无统计学差异(香农指数:有效组3. 69±0. 61,无效组3. 15±1. 01,P=0. 248),但无效组的乳杆菌属(无效组0. 0015±0. 0000,有效组0. 0141±0. 0002,P=0. 010)和双歧杆菌属(无效组0. 0178±0. 0005,有效组0. 1716±0. 0382,P=0. 011)相对丰度显著低于有效组,而志贺菌属(无效组0. 4161±0. 0750,有效组0. 1093±0. 0173,P=0. 008)和普雷沃氏菌属类群9 (无效组0. 0176±0. 0004,有效组0. 0018±0. 0000,P=0. 044)的相对丰度则显著高于有效组。结论中重度UC患者肠道菌群的α多样性及物种组成可能与是否激素暴露无关,但治疗前肠道菌群的物种组成可能是静脉激素疗效的潜在预测指标。
Objective The aim of this study was to analyze the difference of intestinal microbiota in patients with moderately severe ulcerative colitis( UC) treated with or without glucocorticoid( GC),and explore the predictive value for the response to intravenous GC medication. Methods The clinical data of outpatients and inpatients with moderately severe UC treated between November 1,2016 and June 30,2018 in the Department of Gastroenterology,Peking Union Medical College Hospital were retrospectively collected and analyzed. Patients were divided into GC-exposed group and non GC-exposed group based on whether they were exposed to GC when the fecal sample was collected. Patients from non GC-exposed group who received a full-dose of intravenous GC treatment were divided into GC-effective group and GC-refractory group according to the response after the 3-day treatment. The intestinal microbiota from fecal samples of the UC patient was detected by 16 S rRNA gene amplicon sequencing method. α diversity was estimated using the Shannon index. Metastats analysis was employed in multiple comparisons of the microbiota composition. Results Totally 35 moderately severe UC patients were enrolled in this study,among which 20 were non GC-exposed and the other 15 were GC-exposed. Thirteen patients from the non GC-exposed group received a full-dose of intravenous GC treatment after the collection of fecal samples,and 8 of them were GC-effective,while the other 5 were GC-refractory. Patients exposed to GC or not showed little difference in intestinal microbiota α divesity( Shannon index: non GC-exposed group 3. 57±0. 73,GC-exposed group 3. 03 ± 1. 15,P = 0. 123) or microbiota composition. Compared with the GC-effective group( Shannon index 3. 69±0. 61),the microbiota α diversity of the GC-refractory group( Shannon index 3. 15±1. 01)was not significantly different( P = 0. 248). Based on the microbiota composition,the relative abundance of Genus lactobacillus( GC-refractory 0. 0015 ± 0. 0000, GC-effective 0. 0141 ± 0. 0002, P = 0. 010) and Genus bifidobacterium( GC-refractory 0. 0178 ± 0. 0005,GC-effective 0. 1716 ± 0. 0382,P = 0. 011) was significantly lower in the GC-refractory group while that of Genus escherichia-shigella( GC-refractory 0. 4161±0. 0750,GC-effective 0. 1093±0. 0173,P = 0. 008) and Genus prevotella 9( GC-refractory 0. 0176±0. 0004,GC-effective 0. 0018±0. 0000,P = 0. 044) was significantly higher. Conclusions The α diversity or composition of intestinal microbiota of patients with moderately severe UC may not be correlated with GC treatment. It is possible that pre-treated microbiota composition is related to the response to the intravenous GC treatment.

关键词(KeyWords)： 溃疡性结肠炎;肠道菌群;糖皮质激素
ulcerative colitis;intestinal microbiota;glucocorticoid

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金(81770559);; 中国医学科学院医学与健康科技创新工程(2016-12M-3-001)

作者(Author): 李映荷;孟祥辰;阎鹏光;杨红;李骥;钱家鸣;李景南;
LI Ying-he;MENG Xiang-chen;YAN Peng-guang;YANG Hong;LI Ji;QIAN Jia-ming;LI Jing-nan;Department of Gastroenterology,Peking Union Medical College Hospital,Chinese Academy of Medical Sciences & Peking Union Medical College;Laboratory of Intestinal Microecology and Clinical Translational Research,Chinese Academy of Medical Sciences;

Email:

DOI:

参考文献(References)：

• [1]Xavier RJ,Podolsky DK.Unravelling the pathogenesis of inflammatory bowel disease[J].Nature,2007,448:427-434.
• [2]Yang H,Li Y,Wu W,et al.The Incidence of Inflammatory Bowel Disease in Northern China:A Prospective PopulationBased Study[J].PloS ONE,2014,9:e101296.
• [3]Hormannsperger G,Schaubeck M,Haller D.Intestinal microbiota in animal models of inflammatory diseases[J].ILAR J,2015,56:179-191.
• [4]Magnusson MK,Strid H,Isaksson S,et al.The Mucosal antibacterial response profile and fecal microbiota composition are linked to the disease course in patients with newly diagnosed ulcerative colitis[J].Inflamm Bowel Dis,2017,23:956-966.
• [5]Mar JS,LaMere BJ,Lin DL,et al.Disease severity and immune activity relate to distinct interkingdom gut microbiome states in ethnically distinct ulcerative colitis patients[J].MBio,2016,7:e01072-16.
• [6]Walujkar SA,Kumbhare SV,Marathe NP,et al.Molecular profiling of mucosal tissue associated microbiota in patients manifesting acute exacerbations and remission stage of ulcerative colitis[J].World J Microb Biot,2018,34:76.
• [7]Morgan XC,Tickle TL,Sokol H,et al.Dysfunction of the intestinal microbiome in inflammatory bowel disease and treatment[J].Genome Biol,2012,13:R79.
• [8]Kabeerdoss J,Jayakanthan P,Pugazhendhi S,et al.Alterations of mucosal microbiota in the colon of patients with inflammatory bowel disease revealed by real time polymerase chain reaction amplification of 16S ribosomal ribonucleic acid[J].Indian J Med Res,2015,142:23-32.
• [9]Michail S,Durbin M,Turner D,et al.Alterations in the gut microbiome of children with severe ulcerative colitis[J].Inflamm Bowel Dis,2012,18:1799-1808.
• [10]Magnusson MK,Strid H,Sapnara M,et al.Anti-TNFtherapy response in patients with ulcerative colitis is associated with colonic antimicrobial peptide expression and microbiota composition[J].J Crohns Colitis,2016,10:943-952.
• [11]Shaw SA,Bertha M,Hofmekler T,et al.Dysbiosis,inflammation,and response to treatment:a longitudinal study of pediatric subjects with newly diagnosed inflammatory bowel disease[J].Genome Med,2016,8:75.
• [12]Shah R,Cope JL,Nagy-Szakal D,et al.Composition and function of the pediatric colonic mucosal microbiome in untreated patients with ulcerative colitis[J].Gut Microb,2016,7:384-396.
• [13]中华医学会消化病学分会炎症性肠病学组.炎症性肠病诊断与治疗的共识意见(2018年,北京)[J].中华消化杂志,2018,38:292-311.
• [14]Schroeder KW,Tremaine WJ,Ilstrup DM.Coated oral5-aminosalicylic acid therapy for mildly to moderately active ulcerative colitis.A randomized study[J].N Engl J Med,1987,317:1625-1629.
• [15]Satsangi J,Silverberg MS,Vermeire S,et al.The Montreal classification of inflammatory bowel disease:controversies,consensus,and implications[J].Gut,2006,55:749-753.
• [16]Seah D,De Cruz P.Review article:the practical management of acute severe ulcerative colitis[J].Aliment Pharm Ther,2016,43:482-513.
• [17]Mumbi MP,Eissa N,Noah BC.Antepartum antibiotic treatment increases offspring susceptibility to experimental colitis:a role of the gut microbiota[J].PloS ONE,2015,10:e0142536.
• [18]Edgar RC.UPARSE:highly accurate OTU sequences from microbial amplicon reads[J].Nat Methods,2013,10:996-998.
• [19]Cole JR,Wang Q,Cardenas E,et al.The Ribosomal Database Project:improved alignments and new tools for rRNAanalysis[J].Nucleic Acids Res,2009,37:D141-D145.
• [20]Wang M,AhrnéS,Antonsson M,et al.T-RFLP combined with principal component analysis and 16S rRNA gene sequencing:an effective strategy for comparison of fecal microbiota in infants of different ages[J].J Microbiol Methods,2004,59:53-69.
• [21]White JR,Nagarajan N,Pop M.Statistical methods for detecting differentially abundant features in clinical metagenomic samples[J].PLoS Comput Biol,2009,5:e1000352.
• [22]Clarke KR.Non-parametric multivariate analysis of changes in community structure[J].Aust J Ecol,1993,18:117-143.
• [23]Durchschein F,Petritsch W,Hammer HF.Diet therapy for inflammatory bowel diseases:The established and the new[J].World J Gastroenterol,2016,22:2179-2194.
• [24]Cattaneo A,Cattane N,Galluzzi S,et al.Association of brain amyloidosis with pro-inflammatory gut bacterial taxa and peripheral inflammation markers in cognitively impaired elderly[J].Neurobiol Aging,2017,49:60e68.
• [25]Vega-Maga1a N,Delgado-Rizo V,García-Benavides L,et al.Bacterial translocation is linked to increased intestinal IFN-γ,IL-4,IL-17,and mucin-2 in cholestatic rats[J].Ann hepatol,2018,17:318-329.
• [26]Larsen JM.The immune response to prevotella bacteria in chronic inflammatory disease[J].Immunology,2017,151:363-374.
• [27]De Iudicibus S,Franca R,Martelossi S,et al.Molecular mechanism of glucocorticoid resistance in inflammatory bowel disease[J].World J Gastroenterol,2011,17:1095-1108.
• [28]Wine E,Mack DR,Hyams J,et al.Interleukin-6 is associated with steroid resistance and reflects disease activity in severe pediatric ulcerative colitis[J].J Crohns Colitis,2013,7:916-922.
• [29]Santaolalla R,Ma1éJ,Pedrosa E,et al.Apoptosis resistance of mucosal lymphocytes and IL-10 deficiency in patients with steroid-refractory Crohn's disease[J].Inflamm Bowel Dis,2011,17:1490-1500.