文章摘要
邱新运,赵小静,毛夏琼,张红杰.长双歧杆菌对克罗恩病患者外周血单核细胞IL⁃10、IL⁃12、TGF⁃β分泌以及CD25+Foxp3+Treg细胞分化的影响[J].南京医科大学学报,2020,(8):1156~1162
长双歧杆菌对克罗恩病患者外周血单核细胞IL⁃10、IL⁃12、TGF⁃β分泌以及CD25+Foxp3+Treg细胞分化的影响
Effects of Bifidobacterium longum on secretion of IL⁃10,IL⁃12 and TGF⁃β and differentiation of CD25+Foxp3+Treg cells in peripheral blood mononuclear cells in patients with Crohn’s disease
投稿时间:2019-06-07  
DOI:10.7655/NYDXBNS20200813
中文关键词: 克罗恩病  长双歧杆菌  外周血单个核细胞  细胞因子  调节T细胞
英文关键词: Crohn’s disease  Bifidobacterium longum  peripheral blood mononuclear cells  cytokines  regulatory T cell
基金项目:国家自然科学基金(81600416,81470827);江苏省自然科学基金(BK20161065)
作者单位
邱新运 南京医科大学第一附属医院消化内科江苏 南京 210029 
赵小静 南京医科大学第一附属医院消化内科江苏 南京 210029 
毛夏琼 南京医科大学第一附属医院消化内科江苏 南京 210029 
张红杰 南京医科大学第一附属医院消化内科江苏 南京 210029 
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中文摘要:
      目的:比较克罗恩病(Crohn’s disease,CD)患者与健康人群肠道菌群的丰度,研究长双歧杆菌对CD患者外周血单核细胞(peripheral blood mononuclear cell,PBMC)中CD25+Foxp3+Treg细胞分化以及白细胞介素(interleukin,IL)?10、IL?12和转化生长因子(transforming growth factor,TGF)?β分泌的影响。方法:收集19例活动期CD患者和20例健康对照人群的粪便,对16S rRNA片段进行高通量测序。将长双歧杆菌、长双歧杆菌上清、长双歧杆菌培养基或PBS分别与CD患者PBMC体外共培养后,采用流式细胞术检测外周血中CD25+Foxp3+Treg细胞比例;用ELISA法检测细胞培养上清液中IL?10、IL?12和TGF?β 浓度。结果:CD患者肠道菌群与健康人群有显著差别。门水平上,CD患者组变形菌门含量显著增加,而厚壁菌门和放线菌门含量显著减少;属水平上,CD患者组大肠杆菌?志贺氏菌、链球菌及韦荣氏球菌属含量显著增加,而柔嫩梭菌属、Gemmiger、双歧杆菌、瘤胃球菌、罗斯氏菌和Fusicatenibacter等菌属的含量显著降低。与培养基和PBS组相比,长双歧杆菌及其上清液均可刺激CD患者PBMC分泌大量抗炎因子IL?10和TGF?β,而对促炎因子IL?12并没有显著的刺激作用。长双歧杆菌及其上清液组IL?10/IL?12比值及CD25+Foxp3+Treg细胞分化比例较培养基组和PBS组明显增高,其中长双歧杆菌上清液组增高最为明显。结论:长双歧杆菌及其上清液均显示出抗炎作用,并且能够诱导外周血中CD25+Foxp3+Treg的分化。长双歧杆菌的代谢产物可能在未来炎症性肠病治疗中起着重要作用。
英文摘要:
      Objective:To investigate the change of bacterial composition in the gut of Crohn’s disease(CD)patients in comparison with the healthy subjects and the effects of Bifidobacterium longum(B. longum) on differentiation of peripheral blood mononuclear cells(PBMCs) to CD25+Foxp3+Treg cells and secretion of interleukin(IL)?10,IL?12,and transforming growth factor(TGF)?β of PBMCs in patients with CD. Methods:Fecal samples were obtained from active CD patients(n=19)and healthy subjects(n = 20). Bacterial microbiome was analyzed by high?throughput sequencing of 16S rRNA. PBMCs obtained from CD patients were co?cultured in vitro with B. longum,B. longum supernatant,B. longum medium and PBS. The proportion of CD25+Foxp3+Treg cells in the peripheral blood was determined by flow cytometry. Concentrations of IL?10,IL?12 and TGF?β in culture supernatant were measured by ELISA. Results:We observed that bacterial microbiota was skewed in CD,with an increased proportion of Proteobacteria and a decreased proportion of Firmicutes and Actinobacteria at the phylum level,and an increased proportion of Escherichia Shigella,Streptococcus,and Veillonella,while a decreased proportion of Faecalibacterium,Gemmiger,Bifidobacterium,Ruminococcus,Roseburia and Fusicatenibacter at the genus level. Compared with B. longum medium and PBS,B. longum and B. longum supernatant showed a significant higher induction capacity of the anti?inflammatory cytokine IL?10 and TGF?β,while did not remarkably elicit the production of pro?inflammatory cytokine IL?12. The B. longum supernatant induced the highest IL?10/IL?12 ratio and the frequency of peripheral blood CD25+Foxp3+Treg cells in PBMCs,followed by the B. longum. Conclusion:Both B. longum and B. longum supernatant display anti?inflammatory capacities and properties to induce the peripheral blood CD25+Foxp3+Treg cells differentiation. The metabolite of B. longum shows a promising role in treating the inflammatory bowel disease.
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