类风湿关节炎患者肠道菌群分析与外周血细胞因子关系的研究

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目的:通过检测RA患者粪便中菌群种类及分布情况,揭示肠道菌群变化与RA发病的联系,为临床治疗提供新思路。方法:基于16S rDNA高通量测序平台对111例RA患者粪便标本中的16S rDNA V3区进行分析,与100名健康成人粪便标本中菌群的多样性与丰度统计数据进行比较,使用mothur软件分析Alpha多样性,采用Mann-Whitney检验筛选差异菌群,同时分析肠道菌群和外周血细胞因子的关系。结果:2组研究对象性别( n χ2=0.005,n P=0.947)、年龄(n t=0.728,n P=0.467)比较差异均无统计学意义。RA组样品显示物种丰富度的Chao1指数(n Z=-2.188,n P=0.029)和ACE指数(n Z=-2.078,n P=0.038)以及多样性指数Shannon指数(n Z=-2.064,n P=0.039)和Simpsion指数(n Z=-2.064,n P=0.039)均低于健康对照组。在属的水平上,RA组中n Bifidobacterium(n Z=-2.388,n P=0.017)、n Lactobacillus(n Z=-2.543,n P=0.011)、n Clostridium sensu stricto(n Z=-3.842,n P<0.01)、n Blautia(n Z=-2.064,n P=0.039)、n Clostridium Ⅺ(n Z=-2.682,n P<0.01)、n Turicibacter(n Z=-2.437,n P=0.015)、n Phascolarctobacterium (n Z=-3.524,n P<0.01)、n Megasphaera (n Z=-2.870,n P<0.01)、n Veillonella (n Z=-2.472,n P=0.013)、n Citrobacter (n Z=-3.263,n P<0.01)和n Escherichia/n Shigella (n Z=-4.265,n P<0.01)相对丰度较健康对照组明显增加,n Butyricimonas (n Z=-3.071,n P<0.01)、n Odorbacter (n Z=-2.257,n P=0.024)、n Blautia(n Z=-2.064,n P=0.039)、n Clostridium ⅩⅣb (n Z=-2.901,n P<0.01)、n Lachnospiracea incertae sedis(n Z=-2.159,n P=0.031)、n Acetivibrio(n Z=-2.995,n P<0.01)、n Butyricicoccus (n Z=-2.162,n P=0.031)和n Gemmiger(n Z=-2.949,n P±4)。同时,高水平IL-17和高水平TNF-α患者比低水平患者的Prevotella相对丰度增加。n 结论:RA患者肠道微生态环境中细菌群落的多样性和丰富度与健康人群差异有统计学意义,高水平炎症细胞因子的患者Prevotella丰度增加,提示肠道菌群异常与RA中的炎症反应有关,肠道菌群失调可能参与RA的发生发展。“,”Objective:To detect the characteristics of bacteria in the feces of patients with rheumatoid arthritis (RA) and to further discover the relationship between intestinal flora and the status of peripheral cytokine, which might be able to provide new ideas for clinical treatment.Methods:The bacterial diversity and abundance of 111 RA patients and 100 age-and gender-matched healthy controls (HC) were detected by 16S high-throughput sequencing platform and compared. Based on the 16S rDNA high-throughput sequencing platform, the 16S rDNA V3 region in the participants' fecal specimens were analyzed and compared to screen for different bacterial groups. Alpha diversity was analyzed by the mothur software and the screening for different flora was tested by using Mann-Whitney, and the relationship between intestinal flora and peripheral cytokines were analyzed, too.Results:There was no significant difference in gender ( n χ2=0.005, n P=0.947) and age (n t=0.728, n P=0.467) between the two groups. Patients with RA had a lower chao1 index (n Z=-2.188, n P=0.029) and ACE index (n Z=-2.078, n P=0.038) of species richness, and the Shannon index (n Z=-2.064,n P=0.039) and Simpion index (n Z=-2.064, n P=0.039) of diversity index in the feces compared with those of HC. At the genus level, the relative abundance of n Bifidobacterium (n Z=-2.388, n P=0.017), n Lactobacillus (n Z=-2.543, n P=0.011), n Clostridium sensu stricto (n Z=-3.842, n P<0.01),n Blautia (n Z=-2.064, n P=0.039)n , Clostridium Ⅺ (n Z=-2.682, n P<0.01),n Turicibacter (n Z=-2.437, n P=0.015), n Phascolarctobacterium (n Z=-3.524, n P<0.01),n Megasphaera (n Z=-2.87, n P<0.01),n Veillonella (n Z=-2.472, n P=0.013), n Citrobacter (n Z=-3.263, n P<0.01) andn Escherichia/Shigella (n Z=-4.265, n P<0.01) in RA were significantly higher than those of HC (n P<0.05),n Butyricimonas (n Z=-3.071, n P=0.002), n Odorbacter (n Z=-2.257, n P=0.024), n Blautia (n Z=-2.064, n P=0.039), n Clostridium_ⅩⅣb (n Z=-2.901, n P<0.01),n Lachnospiracea_incertae sedis (n Z=-2.159, n P=0.031), n Acetivibrio (n Z=-2.995, n P<0.01),n Butyricicoccus (n Z=-2.162, n P=0.031) and n Gemmiger (n Z=-2.949, n P<0.01) relative abundance were significantly decreased in RA patients (n P<0.05). LEfSe analysis showedn γ-proteobacteria and n Lachnospiraceaehad the most significant difference between the two groups. Further, patients with high inflammatory cytokines such as IL-17 and TNF-α hada higher relative abundance of Prevotella.n Conclusion:The diversity and abundance of intestinal flora in RA patients are significantly different from those of healthy population, which is closely related to the levels of inflammatory cytokines, suggesting imbalance of intestinal flora might be involved in the occurrence and development of RA.
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