中华皮肤科杂志 ›› 2020, Vol. 53 ›› Issue (9): 692-697.doi: 10.35541/cjd.20200208

• 论著 • 上一篇    下一篇

基于16S rRNA基因测序的寻常型银屑病患者肠道菌群差异研究

王丽玮    徐浩翔    段志敏    崔盘根    龚春燕    李岷   

  1. 中国医学科学院  北京协和医学院  皮肤病医院皮肤科,南京  210042
  • 收稿日期:2020-03-05 修回日期:2020-07-04 发布日期:2020-08-31
  • 通讯作者: 李岷 E-mail:drlimin@sina.cn
  • 基金资助:
    中国医学科学院医学与健康科技创新工程项目(2017?12M?1?017)

Analysis of differences in intestinal microbiome in patients with psoriasis vulgaris by using 16S rRNA gene sequencing

Wang Liwei, Xu Haoxiang, Duan Zhimin, Cui Pangen, Gong Chunyan, Li Min   

  1. Department of Dermatology, Hospital for Skin Diseases, Chinese Academy of Medical Sciences and Peking Union Medical College, Nanjing 210042, China
  • Received:2020-03-05 Revised:2020-07-04 Published:2020-08-31
  • Contact: Li Min E-mail:drlimin@sina.cn
  • Supported by:
    CAMS Innovation Fund for Medical Sciences (2017?12M?1?017)

摘要: 【摘要】 目的 研究成年寻常型银屑病患者和健康人群之间肠道微生物的差异。方法 采集2017年9月至2018年2月于中国医学科学院皮肤病医院确诊的22例寻常型银屑病患者和23例健康对照者粪便样本,提取肠道菌群总DNA扩增,并采用二代16S rRNA基因靶向测序技术测序,分析菌群多样性及菌群分布。对照Silva数据库进行物种注释及分类,采用秩和检验分析各层级样本物种差异;采用QIIME软件(Version 1.9.1)计算OTU数目及α多样性主要指数(Chao1指数、Shannon指数和Simpson指数),t检验分析指数差异;PCoA分析反映样本β多样性差异,置换多元方差分析两组群落组成结构差异;秩和检验及LEfSe分析评估物种差异。结果 银屑病组OTU数目(147.55 ± 57.07)与健康对照组(148.96 ± 50.45)比较,差异无统计学意义(t = 0.088,P = 0.930)。银屑病组Shannon指数(4.08 ± 0.80)、Chao1指数(169.52 ± 63.17)、Simpson指数(0.87 ± 0.07)与健康对照组(分别为4.11 ± 0.94、175.36 ± 53.59、0.86 ± 0.90)比较,差异均无统计学意义(t = 0.12、0.34、0.27,均P > 0.05)。PCoA分析银屑病组与健康对照组的第一主成分差异为49.8%,第二主成分差异为15.62%,置换多元方差分析显示两组间β多样性差异有统计学意义(P = 0.011)。银屑病组与健康对照组有差异的菌群包括在银屑病组中显著升高的厚壁菌门、梭菌纲、梭菌目,丹毒丝菌目、丹毒丝菌科,健康对照组中显著升高的艾普西隆变形杆菌纲、弯曲杆菌目、弯曲杆菌科、弯曲杆菌属,拟杆菌目、拟杆菌科。结论 寻常型银屑病患者与健康人的肠道菌群存在一定差异,肠道菌群有可能成为寻常型银屑病的潜在生物标志物。

关键词: 银屑病, 肠道, 生物群, RNA, 核糖体, 16S, 基因测序

Abstract: 【Abstract】 Objective To investigate differences in intestinal microbiome between adult patients with psoriasis vulgaris and healthy individuals. Methods Fecal samples were collected from 22 patients with confirmed psoriasis vulgaris and 23 healthy controls in Hospital for Skin Diseases, Chinese Academy of Medical Sciences and Peking Union Medical College from September 2017 to February 2018. The total DNA of intestinal flora was extracted and amplified, and the next-generation 16S rRNA gene-targeted sequencing was performed to analyze the diversity and distribution of intestinal flora. Species annotation and classification were performed according to Silva database, and rank sum test was used to analyze species differences among samples at different taxonomic ranks; QIIME software (Version 1.9.1) was used to calculate the number of operational taxonomic units (OTUs) and main indices of α diversity (Chao1 index, Shannon index and Simpson index), and t test to analyze differences in indices; PCoA analysis was performed to analyze the difference in β diversity, and differences in microbial community composition structure were analyzed between the two groups by using permutational multivariate analysis of variance; rank sum test and linear discriminant analysis effect size (LEfSe) analysis were used to evaluate the species difference. Results No significant difference in the number of OTUs was observed between the psoriasis group (147.55 ± 57.07) and healthy control group(148.96 ± 50.45, t = 0.088, P = 0.930). In addition, there were no significant differences in the Shannon index, Chao1 index or Simpson index between the psoriasis group (4.08 ± 0.80, 169.52 ± 63.17, 0.87 ± 0.07, respectively) and healthy control group (4.11 ± 0.94, 175.36 ± 53.59, 0.86 ± 0.90, respectively; t = 0.12, 0.34, 0.27, all P > 0.05). PCoA analysis showed that the first and second principal components explained 49.8% and 15.62%, respectively, of the total variance between the psoriasis group and healthy control group, and permutational multivariate analysis of variance revealed that the β diversity significantly differed between the two groups (P = 0.011). Different microbes between the psoriasis group and healthy control group included Firmicutes, Clostridia, Clostridiales, Erysipelotrichales and Erysipelotrichaceae, whose abundance significantly increased in the psoriasis group, as well as Epsilonproteobacteria, Campylobacterales, Campylobacteraceae, Campylobacter, Bacteroidales and Bacteroidaceae, whose abundance significantly increased in the healthy control group. Conclusion The intestinal microbiome differs between patients with psoriasis vulgaris and healthy individuals, which may serve as potential biomarkers for psoriasis vulgaris.

Key words: Psoriasis, Intestines, Biota, RNA, ribosomal, 16S, Gene sequencing