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在单次根除事件前后,幽门螺杆菌感染患者的胃肠道微生物种群结构无显著差异。

Lack of significant differences between gastrointestinal tract microbial population structure of Helicobacter pylori-infected subjects before and 2 years after a single eradication event.

机构信息

Latvian Biomedical Research and Study Centre, Riga, Latvia.

Institute of Clinical and Preventive Medicine, University of Latvia, Riga, Latvia.

出版信息

Helicobacter. 2020 Oct;25(5):e12748. doi: 10.1111/hel.12748. Epub 2020 Aug 9.

Abstract

BACKGROUND

According to recent estimates 80% of Latvian population is infected with Helicobacter pylori thus their susceptibility to numerous gastric tract diseases is increased. The 1 line H. pylori eradication therapy includes treatment with clarithromycin in combination with amoxicillin or metronidazole and a proton pump inhibitor. However, potential adverse events caused by such therapies to microbiome are insufficiently studied.

OBJECTIVE

This study aimed to evaluate the long-term effect of H. pylori eradication on human gastrointestinal tract (GIT) microbiome.

METHODS

The assessment of H pylori eradication impact on GIT microbiome was done by analyzing 120 samples acquired from 60 subjects. Each individual was prescribed the following 10-day eradication regimen: Esomeprazolum 40 mg, Clarithromycinum 500 mg, and Amoxicillinum 1000 mg, BID. Samples from each individual were collected before starting H pylori eradication therapy, and 2 years after the completion of the therapy in OC-Sensor (Eiken Chemical Co.) sample collection containers and stored at -86°C. Prior to DNA extraction, the samples were lyophilized, and total DNA was extracted using FastDNA Spin Kit for Soil. 16S V3 rRNA gene sequencing was done employing Ion Torrent PGM, and the obtained raw sequences were analyzed using vsearch and R (phyloseq, cluster packages).

RESULTS

Alpha diversity measurements-observed OTUs, Chao1 and Shannon index did not differ significantly between the pre- and post-eradication states (two-tailed paired t test: P = .95; P = .71, P = .24, respectively). Unweighted and weighted UniFrac distances of beta diversity analysis indicated a non-specific pattern of sample clustering. Enterotype shift was observed for the majority of individuals comparing pre- and post-eradication study groups. Association analysis revealed that certain bacterial genera significantly correlated with age (eg, Dialister, Paraprevotella, Bifidobacterium), individual (eg, Thermotunica, Streptomyces, Faecalibacterium), and history of respiratory and/or allergic diseases (eg, Colinsella, Faecalibacterium). Redundancy analysis confirmed that the individual was a significant determinant of the subject's microbial community composition (ANOVA, 999 perm., P = .001) with the further lower impact of subject-specific medical history (eg, medication used as prescribed: P = .005, history of cardiovascular diseases: P = .005, history of respiratory, and/or allergic diseases: P = .015) and physiological (eg, age: P = .005, gender: P = .02) parameters. In the post-eradication study group, number of influential genera (n = 260) was increased compared to the pre-eradication study group (n = 209).

CONCLUSION

Modest global differences at the community level exist between individuals before and after the eradication therapy; however, the microbiome structure is more related to the subject-specific parameters rather than by the eradication therapy itself.

摘要

背景

根据最近的估计,80%的拉脱维亚人感染了幽门螺杆菌,因此他们患许多胃肠道疾病的风险增加。一线 H. pylori 根除疗法包括使用克拉霉素联合阿莫西林或甲硝唑和质子泵抑制剂进行治疗。然而,这种疗法对微生物组的潜在不良反应尚未得到充分研究。

目的

本研究旨在评估 H. pylori 根除对人类胃肠道 (GIT) 微生物组的长期影响。

方法

通过分析 60 名受试者的 120 个样本,评估 H. pylori 根除对 GIT 微生物组的影响。每位个体均接受以下 10 天的根除治疗方案:埃索美拉唑 40mg、克拉霉素 500mg 和阿莫西林 1000mg,每日 2 次。在开始 H. pylori 根除治疗之前和治疗完成后 2 年,从每个个体中采集样本,并在 OC-Sensor(Eiken Chemical Co.)样本采集容器中储存于-86°C。在提取 DNA 之前,将样本冻干,使用 FastDNA Spin Kit for Soil 提取总 DNA。使用 Ion Torrent PGM 进行 16S V3 rRNA 基因测序,并用 vsearch 和 R(phyloseq、cluster 包)分析获得的原始序列。

结果

在根除治疗前后,α多样性测量——观察到的 OTUs、Chao1 和 Shannon 指数没有显著差异(双侧配对 t 检验:P=0.95;P=0.71,P=0.24)。β多样性分析的无权重和加权 UniFrac 距离表明样本聚类没有特异性模式。大多数个体在根除前后的研究组中观察到肠型转变。关联分析表明,某些细菌属与年龄(例如,Dialister、Paraprevotella、Bifidobacterium)、个体(例如,Thermotunica、Streptomyces、Faecalibacterium)和呼吸及/或过敏疾病史(例如,Collinsella、Faecalibacterium)显著相关。冗余分析证实个体是个体微生物群落组成的重要决定因素(ANOVA,999 次 perm.,P=0.001),个体特有的医学史(例如,按规定使用药物:P=0.005,心血管疾病史:P=0.005,呼吸及/或过敏疾病史:P=0.015)和生理参数(例如,年龄:P=0.005,性别:P=0.02)的影响较低。在根除治疗后的研究组中,有影响力的属的数量(n=260)比根除治疗前的研究组(n=209)有所增加。

结论

在根除治疗前后,个体之间在群落水平上存在适度的整体差异;然而,微生物组结构更多地与个体特有的参数相关,而不是与根除治疗本身相关。

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