黄连素对艰难梭菌相关性腹泻小鼠肠道菌群的调节作用研究

摘要

目的 评估黄连素对艰难梭菌相关性腹泻模型(CDAD)小鼠肠道菌群的调节作用.方法50只C57BL/6小鼠按照数字表法随机分为5组,每组10只.空白对照组不给予任何干预.余下4组小鼠经抗生素混合液预处理和克林霉素腹腔注射后给予艰难梭菌108CFU/L灌胃,建立抗生素诱导菌群失调致艰难梭菌相关性腹泻动物模型.造模组造模后不予药物治疗,黄连素组、万古霉素组、联合用药组分别给予黄连素100mg/(kg·d)、万古霉素50mg/(kg·d)及二者联合灌胃治疗.治疗结束后处死小鼠,取回盲部内容物进行Illumina Miseq PE300高通量测序检测菌群变化情况.结果CDAD小鼠模型肠道三大菌门的构成比(厚壁菌门48.52％,拟杆菌门0.19％,变形菌门51.29％,γ-变形菌纲成为优势细菌)与正常对照组(厚壁菌门55.55％、拟杆菌门35.13％、变形菌门8.81％,其中以δ-变形菌纲和ε-变形菌纲为主)相比发生显著改变.万古霉素组厚壁菌门9.72％,拟杆菌门64.00％,变形菌门26.10％,仍以γ-变形菌纲为主.黄连素组厚壁菌门21.95％,拟杆菌门70.72％,变形菌门3.36％,以δ-变形菌纲为优势菌.联合用药组厚壁菌门54.17％,拟杆菌门44.35％,变形菌门1.42％,δ-变形菌纲再次成为优势细菌.结论在艰难梭菌相关性腹泻小鼠模型中,黄连素可显著抑制多形菌门肠杆菌科细菌的繁殖,促进了小鼠肠道菌群的恢复,与万古霉素联合应用可减少万古霉素单独治疗的复发问题.%Objective To evaluated the effect of berberine on regulation gut microflora in the CDAD C57BL/6 mouse model.Method The 40 mice Clostridium dififcile associated diarrhea Model was established by being given a gavage of C. dififcile strain 108 CFU/L after pretreatment with an antibiotic cocktail and Clindamycin Intraperitoneal. The control group 10 mice were treated without any drugs. The other three groups (10 mice/group) were treated seperately with Berberine at 100 mg/(kg?d) through gavage or Vancomycin at 50 mg/(kg?d) or combination of both Vancomycin and Berberine through gavage. After treatment, mice were killed and ileocecus content were obtained. Then Illumina Miseq PE300 sequencing was used to determine the overall structural change of microflora.Results The bacteria structure of CDAD mouse model was obviously different with the Control group (Firmicutes 55.55％, Bacteroidetes 35.13％, Proteobacteria 8.81％,δ-Proteobacteria andε-Proteobacteria are the major), Firmicutes 48.52％, Bacteroidetes 0.19％, Proteobacteria 51.29％ (γ-Proteobacteria is the major) in CDAD model group. Vancomycin group: Firmicutes9.72％, Bacteroidetes64.00％, Proteobacteria 26.10％ (γ-Proteobacteria is major). Berberine group: Firmicutes 21.95％, Bacteroidetes70.72％, Proteobacteria 3.36％ (δ-Proteobactera is major). Conmbination group: Firmicutes 54.17％, Bacteroidetes 44.35％, Proteobacteria 1.42％ (δ-Proteobacteria becmame the major again).Conclusion In the study of CDAD mouse model, Berberine administration significantly promoted the restoration of intestinal microflora by inhibiting the expansion of Proteobacteria/Enterobacteriaceae. Our data indicates that a combination of vancomycin and berberine is more effective than vancomycin alone for treating CDAD.