咽喉反流性疾病患者肠道菌群及代谢组学特征与治疗前后变化分析

Analysis of Gut Microbiota and Metabolomic Characteristics in Patients with Laryngopharyngeal Reflux Disease and Their Changes Before and After Treatment

摘要:

目的：分析咽喉反流性疾病（LPRD）患者以及健康人群的肠道菌群结构及其代谢物特征，明确其与LPRD的相关性，并探究治疗前后肠道菌群及其代谢物变化。 方法：纳入联勤保障部队第九〇〇医院18例LPRD患者（病例组）与14例健康志愿者（对照组），收集临床资料及新鲜粪便样本。采用16S rDNA测序和LC-MS代谢组学技术，分析肠道菌群多样性、差异菌群及差异代谢物，并进行多组学关联分析。病例组治疗前为LPRD-A组，治疗后为LPRD-B组，对照组为C组。 结果：1.微生物组学：α多样性分析显示各组间菌群多样性和丰富度无显著差异（P > 0.05）；β多样性分析显示LPRD-A组和C组、LPRD-B组与C组间菌群结构存在显著差异（P < 0.05），而LPRD-B组与LPRD-A组间无显著差异（P > 0.05）；LEfSe差异分析显示，C组富集厚壁菌门、梭菌纲、巨单胞菌属等；LPRD-A组富集拟杆菌目、伯克氏菌目、柯林斯菌属等；LPRD-B组富集克雷伯氏菌属、肠杆菌属等。2.代谢组学：与C组相比，LPRD-A组中α,α-海藻糖-6-磷酸、L-亮氨酰-L-脯氨酸等代谢物上调，甲硫腺苷等下调；与LPRD-A组相比，LPRD-B组中有机氮化合物等上调，N-乙酰神经氨酸等下调。3.关联分析：LPRD-A组和C 组、LPRD-B组和LPRD-A组在多组学关联中区分明显，分别发现29对和5对显著相关的菌群-代谢物。结论：肠道菌群及其代谢物与LPRD存在相关性。差异菌群及代谢物可能参与LPRD的发生与发展。

Abstract:

Purpose: To analyze the structural characteristics of gut microbiota and their metabolites in patients with laryngopharyngeal reflux disease (LPRD) and healthy individuals, clarify their correlation with LPRD, and investigate the changes in gut microbiota and metabolites before and after treatment.Methods: A total of 18 LPRD patients (case group) and 14 healthy volunteers (control group) were enrolled from the Department of Otorhinolaryngology Head and Neck Surgery, the 900th Hospital of the Joint Logistics Support Force. Clinical data and fresh stool samples were collected. Gut microbiota diversity, differential microbiota, and differential metabolites were analyzed using 16S rDNA sequencing and LC-MS metabolomics, followed by multi-omics association analysis. Samples from the case group before treatment were designated as the LPRD-A group, after treatment as the LPRD-B group, and samples from the control group as group C. Result: 1. Microbiomics: α diversity analysis showed no significant differences in microbial diversity or richness among the groups (P > 0.05). β diversity analysis revealed significant differences in gut microbiota structure between the LPRD-A and C groups, as well as between the LPRD-B and C groups (P < 0.05), but no significant difference was observed between the LPRD-B and LPRD-A groups (P > 0.05). LEfSe analysis indicated that group C was enriched in Firmicutes, Clostridia, and Megamonas; the LPRD-A group was enriched in Bacteroidales, Burkholderiales, and Collinsella; and the LPRD-B group was enriched in Klebsiella, Enterobacter, and Prevotellaceae_NK3B31_group.2. Metabolomics: Compared with group C, α,α-Trehalose 6-phosphate, L-leucyl-I-proline, were significantly up-regulated in group LPRD-A, while S-methyl-5''-thioadenosine significantly down-regulated; Compared with group LPRD-A, 7(z),10(z),13(z),16(z),9(z)-N-(2-hydroxyethyl)docosapentaenamide, Neohesperidose were significantly upregulated in group LPRD-B, while N-acety|neuraminic acid, N-acetyl-d-glucosamine were significantly down-regulated. 3. Multi-omics association analysis: There were significantly differentiated in Multi-omics association analysis between group LPRD-A and group C, group LPRD-B and group LPRD-A. 29 pairs and 5 pairs of significantly correlated microbiota - metabolites were found respectively. Conclusion: The intestinal flora and its metabolites are correlated with LPRD. Differential microbiota and metabolites may play a role in the onset and progression of the disease.