摘要
目的观察过敏性紫癜(HSP)患儿肠黏膜屏障功能变化,探讨肠道菌群、L/M与Treg/Th17作用关系。方法采用细菌16S r DNA、液相色谱技术(HPLC)、流式细胞术(FCM)、酶联免疫法(ELISA)检测41例HSP急性期患儿、35例HSP临床缓解期患儿,及30例同期健康体检儿粪便中双歧杆菌、乳酸杆菌、大肠埃希菌和肠球菌的基因拷贝数,尿液中乳果糖(Lactulose)和甘露醇(Mannitol)浓度比值(L/M值),以及CD4+CD25+T细胞(Treg)和CD3+CD8-IL-17+(Th17)细胞百分比,及相应转化生长因子β1(TGF-β1)、白细胞介素-17(IL-17)水平。结果 HSP患儿急性期存在肠道菌群紊乱,表现在肠道中乳酸杆菌、双歧杆菌基因拷贝数降低,以乳酸杆菌减少为著,大肠埃希菌和肠球菌减少不明显;外周血单个核细胞(PBMC)中Th17百分数及血浆中IL-17水平增高,Treg细胞及TGF-β1水平降低,Treg/Th17失衡;尿液中L/M值增高。临床缓解期时患儿肠道双歧杆菌、乳酸杆菌基因拷贝数增加,以双歧杆菌上升为著,但肠道菌群结构仍未扶正;PBMC中Th17细胞百分数及血浆中IL-17水平降低,Treg细胞及TGF-β1水平逐渐升高,仍存在Treg/Th17失衡。肠道菌群中双歧杆菌/大肠埃希菌(B/E)比值与Treg/Th17呈正相关关系,与尿液中L/M值呈负相关。结论 HSP患儿急性期肠黏膜屏障功能受损,表现在机械、免疫屏障降低及肠道微生态失衡,肠道菌群所诱导的免疫耐受被打破,并贯穿HSP整个病理过程。
Objective To observe the changes of gut mucosal barrier function and the relationship among intestinal flora,L / M and Treg / Th17 in children with Henoch-Schonlein purpura. Methods Forty-one children with HenochSchonlein purpura（ HSP） were included,with thirty healthy children as the controls. The percentages of CD4^＋CD25^＋T cell（ Treg） and CD3^＋CD8^-IL-17＋（ Th17） cell were detected with flow cytometrie（ FCM）; The levels of TGF-β1 and IL-17 in plasma were detected with enzyme-linked immunosorbent assay. 16 S r DNA fluorescent quantitative PCR was used to quantify Lactobacillus,Bifidobacterium,E. coli and Enterococcus in feces. High performance liquid chromatography was used to analyze the levels of lactulose（ L） and mannitol（ M）,and the intestinal permeability was evaluated based on L / M. Results（ 1） The gene copy numbers of Lactobacillus and Bifidobacterium decreased,especially Lactobacillus,while those of E. coli and Enterococcus did not change significantly,suggesting that alteration of intestinal flora occurred in children with HSP during the acute stage.（ 2） The percentage of Th17 in peripheral blood mononuclear cells and the level of IL-17 in plasma increased while Treg cells and cytokine TGF-β1 decreased during the acute stage,indicating that Treg / Th17 balance was destroyed in the acute stage of HSP. The ratio of lactulose / mannitol（ L / M） increased during the acute stage of HSP.（ 3） In the remission stage,the gene copy number of Bifidobacteria increased more quickly than others,however the overall gut flora structure did not recovered completely in recovery stage. The percentage of Th17 and its cytokine increased,while the percentage of Treg cells and its cytokine in plasma decreased during the recovery stage. Imbanlance of Treg /Th17 still existed. B / E ratio showed positive correlation with Treg / Th17 ratio,and negative correlation with L / M ratio. Conclusion During the acute stage of HSP,there is gut mucosal barrier dysfunction,including dysfun
出处
《中国微生态学杂志》
CAS
CSCD
2015年第9期1022-1026,共5页
Chinese Journal of Microecology
基金
河北省科技厅科技计划项目(13277763D)
