基于高通量测序研究奥狄氏括约肌松弛的胆结石患者胆汁细菌多样性

Bile bacterial diversity in patients with cholelithiasis and the sphincter of Oddi laxity based on high-throughput sequencing

摘要:

为临床治疗奥狄氏括约肌松弛（the Sphincter of Oddi laxity, SOL）患者胆结石复发寻找依据,本文研究了患有SOL的胆结石患者的胆汁细菌组成和多样性. 分析了SOL与非SOL胆结石患者术前肝功、肾功和血常规等临床指标差异，基于Illumina MiSeq平台对患者胆汁细菌进行16S rRNA基因高通量测序. 结果发现，SOL患者的总胆红素（TBIL）、直接胆红素（DBIL）、间接胆红素（IBIL）水平显著高于非SOL患者, 而腺苷脱氨酶（ADA）则显著低于后者（P < 0.05）.两组患者之间胆汁细菌α多样性指数除均匀度外无显著差异（P>0.05）；Wilcoxon秩和检验显示SOL患者胆汁中细菌丰度前四的菌门依次为变形菌门（Proteobacteria）、厚壁菌门（Firmicutes）、放线菌门（Actinobacteria）和拟杆菌门（Bacteroidete），其中变形菌门、肠球菌属（Enterococcus）和Lachnoclostridiums属细菌在SOL患者胆汁中含量显著高于非SOL患者（P< 0.05），而SOL患者中其他优势菌属如克雷伯菌（Klebsiella）、马赛菌属（Massilia）和假单胞杆菌（Pseudomonas）丰度则无显著增高（P>0.05）. 因此，SOL患者的部分临床指标如胆红素、ADA水平和胆汁细菌组成与非SOL患者存在显著差异，这为临床治疗SOL患者胆结石复发提供相关依据.

Abstract:

To study the composition and diversity of bile bacteria in patients with the sphincter of Oddi laxity (SOL), to provide basis for clinical treatment of cholelithiasis recurrence in patients with SOL, this study analyzed the differences of clinical indexes before operation, including liver and kidney function and blood routine index between control (non-SOL patients with cholelithiasis) and SOL group. The high-throughput 16S rRNA gene sequencing of biliary bacteria was performed based on the Illumina MiSeq platform. The results showed that the level of total bilirubin, direct bilirubin and indirect bilirubin in cholelithiasis patients with the SOL were significantly higher than those in the nonSOL group, while adenosine deaminase (ADA) was lower than that in control (P< 0.05). There was no significant difference in bacterial alpha diversity between SOL and non-SOL patients except for evenness (P> 0.05), and Wilcoxon rank-sum test showed that the top four abundant bacterial phyla in SOL patients bile were Proteobacteria, Firmicutes, Actinobacteria and Bacteroidete in turn, among which the abundance of Proteobacteria, Enterococcus and Lachnoclostridium was significantly higher in SOL patients than in control (P< 0.05), so did Klebsiella, Massilia and Pseudomonas (P>0.05). Therefore, there are significant differences in some clinical indexes such as bilirubin, ADA level and bile bacterial composition between patients with SOL and those without SOL, which provide relevant basis for the clinical treatment of cholelithiasis recurrence in patients with SOL.