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初治肺结核患者肠道菌群结构变化与免疫指标的相关性研究

[Correlation study between changes in intestinal microflora structure and immune indexes in newly treated patients with pulmonary tuberculosis].

作者信息

Xie J H, Yu R, Shi G M, Ma X H, Xiao S F, Yi Y H, Zhou T, Xiang Y G

机构信息

Department of Laboratory Medicine,Affiliated Changsha Central Hospital,University of South China, Changsha 410004,China.

出版信息

Zhonghua Yu Fang Yi Xue Za Zhi. 2021 Dec 6;55(12):1486-1490. doi: 10.3760/cma.j.cn112150-20210728-00721.

Abstract

To explore the correlation between the changes of the intestinal flora of newly treated pulmonary tuberculosis patients and the immune indicators of the body, and to provide a reference for the prevention and treatment of pulmonary tuberculosis. A single-center and case-control study was adopted. From October 2020 to April 2021, 43 patients with newly diagnosed tuberculosis in the Department of Tuberculosis, Affiliated Changsha Central Hospital,University of South China were selected as the control group. 43 cases of newly treated pulmonary tuberculosis (PTB), 43 healthy control (HC) during the same period, collected fresh feces and whole blood of subjects, and used Illumina Hiseq high-throughput sequencing technology to analyze 16S of all microorganisms in feces The V4 region of rRNA was amplified and sequenced, and the structure of the intestinal flora was analyzed by QIIME software. Use flow cytometry to determine the subject's immune indicators (CD3, CD4, CD8, CD4CD25CD127Treg, CD14CD16, CD14CD16), and analyze the changes in intestinal flora and immune function in newly treated pulmonary tuberculosis patients Inherent connection. The χ² test, t test, and Wilcox rank sum test were used to analyze the differences in age, gender, α diversity, and relative abundance of the two groups of people. Compared with the HC group, the alpha diversity of the intestinal flora in the PTB group decreased (shannon index: =3.906, =0.000 2; simpson index: =553, =0.004 7; chao1 index: =5.395, =0.000 0). β diversity analysis showed that there were significant differences in the structure of the intestinal flora between the two groups (=0.000). Species difference analysis showed that at the phylum level, the relative abundance of Firmicutes in the PTB group was significantly lower than that in the HC group (=486.0, =0.000 5). At the genus level, there are 15 different bacterial genera between the two groups. In the PTB group, bifidobacterium, enterococcus, lactobacillus, anaerostipes, the relative abundance of the above 5 genera of veillonella is higher than that of the HC group (<0.05); Butyricimonas, clostridium, and broutella (blautia), coprococcus, dorea, lachnospira, roseburia, faecalibacterium, ruminococcus, the relative abundance of 10 bacterial genera including dialister was lower than that of the HC group (<0.05). Comparison of immune indexes between groups showed that CD14CD16monocytes (%) in the PTB group were higher than those in the HC group (=2.456, =0.001 6<0.05), while CD14CD16monocytes (%) were lower than HC (=-4.368, =0.000<0.05), while the differences in CD3, CD4, CD8, CD4/CD8and Treg (CD4CD25CD127) were not statistically significant (>0.05). Spearman correlation analysis showed that Firmicutes in the PTB group was negatively correlated with CD4/CD8, CD14CD16(=-0.218, =0.048; =-0.245, =0.025), and positively correlated with CD14CD16Correlation (=0.250, =0.022); At the genus level, Faecalis is positively correlated with CD4/CD8and CD4(=0.250, =0.023; =0.258, =0.019); Rosella and CD3, CD8and CD14CD16are positively correlated (=0.27, =0.024; =0.219, =0.046; =0.027, =0.039), and negatively correlated with CD14CD16(=-0.280, = 0.01). Changes in the structure of the intestinal flora of newly treated pulmonary tuberculosis patients may be one of the influencing factors of the immune function of the body. Targeted optimization of the structure of the intestinal flora and improvement of the body's immunity may be used as an effective auxiliary treatment for pulmonary tuberculosis.

摘要

探讨初治肺结核患者肠道菌群变化与机体免疫指标之间的相关性,为肺结核的防治提供参考。采用单中心病例对照研究。选取2020年10月至2021年4月在南华大学附属长沙中心医院结核科新诊断的43例肺结核患者作为病例组,同期43例健康对照者及43例初治肺结核患者,采集受试者新鲜粪便和全血,采用Illumina Hiseq高通量测序技术对粪便中所有微生物的16S rRNA的V4区进行扩增测序,用QIIME软件分析肠道菌群结构。采用流式细胞术检测受试者免疫指标(CD3、CD4、CD8、CD4CD25CD127Treg、CD14CD16、CD14CD16),分析初治肺结核患者肠道菌群与免疫功能变化的内在联系。采用χ²检验、t检验及Wilcox秩和检验分析两组人群年龄、性别、α多样性及相对丰度的差异。与健康对照组相比,病例组肠道菌群的α多样性降低(香农指数:=3.906,=0.000 2;辛普森指数:=553,=0.004 7;chao1指数:=5.395,=0.000 0)。β多样性分析显示两组肠道菌群结构存在显著差异(=0.000)。物种差异分析显示,在门水平上,病例组厚壁菌门的相对丰度显著低于健康对照组(=486.0,=0.000 5)。在属水平上,两组间有15种不同的细菌属。病例组中双歧杆菌、肠球菌、乳酸杆菌、厌氧棒状菌、韦荣球菌属这5种菌属的相对丰度高于健康对照组(<0.05);丁酸单胞菌、梭菌属、布劳特氏菌属(blautia)、粪球菌属、多雷阿菌属、毛螺菌属、罗斯氏菌属、粪杆菌属、瘤胃球菌属、戴氏菌属这10种菌属的相对丰度低于健康对照组(<0.05)。组间免疫指标比较显示,病例组CD14CD16单核细胞(%)高于健康对照组(=2.456,=0.001 6<0.05),而CD14CD16单核细胞(%)低于健康对照组(=-4.368,=0.000<0.05),而CD3、CD4、CD8、CD4/CD8及Treg(CD4CD25CD127)差异无统计学意义(>0.05)。Spearman相关性分析显示,病例组厚壁菌门与CD4/CD8、CD14CD16呈负相关(=-0.218,=0.048;=-0.245,=0.025),与CD14CD16呈正相关(=0.250,=0.022);在属水平上,粪肠球菌属与CD4/CD8及CD4呈正相关(=0.250,=0.023;=0.258,=0.019);罗斯氏菌属与CD3、CD8及CD14CD16呈正相关(=0.27,=0.024;=0.219,=0.046;=0.027,=0.039),与CD14CD16呈负相关(=-0.280,=0.01)。初治肺结核患者肠道菌群结构变化可能是机体免疫功能的影响因素之一。针对性优化肠道菌群结构、提高机体免疫力,可作为肺结核的有效辅助治疗手段。

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