银屑病患者咽颊部菌群结构组成分析

Analysis of the composition of pharyngeal buccal flora in psoriasis patients

目的探索银屑病患者咽颊部口腔微生物的组成变化与银屑病的发病以及疾病进展的关系,为银屑病的个性化诊疗提供新的思路。方法采集临床上经过严格筛选的15例银屑病患者(试验组)和15例健康人(对照组)的咽颊部黏膜样本,提取细菌基因组DNA,进行PCR扩增,将扩增后的DNA样本进行16SrRNA高通量测序法分析(Illumina测序)。测序结果与Greengenes Database进行比对,通过生物信息学、医学统计学分析银屑病患者咽颊部口腔微生物结构组成与正常人的差异。结果在菌群门的水平上,试验组中厚壁菌门在数量上占有绝对优势,其余拟杆菌门、变形菌门、放线菌门、梭杆菌门也依次占有一定比例;对照组中厚壁菌门在数量上同样占有绝对优势,拟杆菌门、变形菌门等也占有不小的比例。虽然在相对丰度上银屑病患者与健康对照组有区别,但差异无统计学意义。在菌群属的水平上,试验组患者中颗粒链球菌与梭杆菌占有明显的相对丰度,对照组则以颗粒链球菌、巨型球菌、Bulleidia、Parvimonas、梭杆菌为主。两组菌群在构成上已经表现出明显的区别。另外,在银屑病患者的咽颊部菌群中颗粒链球菌、梭杆菌和Bulleidia的相对丰度有所升高,而Oribacterium与产线菌(Filifactor)则明显降低,与健康人群相比差异有统计学意义(t=2.5010,P<0.05;t=2.0875,P<0.05)。结论银屑病患者与健康正常人咽颊部微生物组成结构在属水平上存在明显的差异。咽颊部微生物在银屑病的发病以及疾病进展中的作用值得深入研究。

Objective To explore the correlation between the composition of pharyngeal buccal flora and pathogenesis of psoriasis, and provide new method for individualized therapy of the disease. Methods Specimens were collected from the pharyngeal buccal mucous membranes of psoriasis patients （n=15） and healthy individuals （n=15） to extract the genome DNA for PCR amplification and Illumina sequencing with high-throughout sequencing technology. The results were compared with the data from Greengenes database to analyse the composition of pharyngeal buccal flora of psoriasis patients vs healthy controls with bioinformatical and medical statistical methods. Results At the level of bacterial phyla, Firmicutes were predominant in psoriasis, followed by Bacteroidetes, Proteobacteria, Actinobacteria and Fusobacteria. In controls Firmicutes were also the most abundant, with Bacteroidetes and Proteobacteria accounting for quite a few percentage. There were differences in the abundance of bacteria between the two groups but with no statistical significance. At the level of bacterial genera, Graulicatella and Fusobacteria were relatively abundant in psoriasis group, while Graulicatella, Megaspheara, Bulleidia, Parvimonasand Fusobacteria were predominant in control group. Moreover, the relative abundances of Ingraulicatella, Fusobacteria and Bulleidia increased while those of Oribacterium and Filifactor decreased in psoriasis patients, significantly different from those in heathy controls （t=2.5010, P〈0.05; t=2.0875, P〈0.05）. Conclusion There are obvious differences in the composition of pharyngeal buccal flora between psoriasis patients and healthy individuals at the level of genera. The roles of pharyngeal buccal microbes in the pathogenesis of psoriasis should be further studied.