Observational Study
Copyright ©The Author(s) 2018. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. Aug 14, 2018; 24(30): 3448-3461
Published online Aug 14, 2018. doi: 10.3748/wjg.v24.i30.3448
Altered oral microbiota in chronic hepatitis B patients with different tongue coatings
Yu Zhao, Yu-Feng Mao, Yi-Shuang Tang, Ming-Zhu Ni, Qiao-Hong Liu, Yan Wang, Qin Feng, Jing-Hua Peng, Yi-Yang Hu
Yu Zhao, Yu-Feng Mao, Yi-Shuang Tang, Ming-Zhu Ni, Qiao-Hong Liu, Yan Wang, Qin Feng, Jing-Hua Peng, Institute of Liver Diseases, Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China
Yi-Yang Hu, Institute of Liver Diseases, Key Laboratory of Liver and Kidney Diseases (Ministry of Education), Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China
Author contributions: Hu YY designed the study; Zhao Y and Tang YS performed the tongue coating microbial experiments; Tang YS, Liu QH and Wang Y performed the tongue coating metabolic experiments; Tang YS, Feng Q and Peng JH performed the clinical trial; Mao YF and Ni MZ performed the statistical analyses; Zhao Y wrote the paper.
Supported by the Shanghai Educational Development Foundation, No. 14CG41; the National Natural Science Foundation of China, No. 81403298 and No. 81373857; and the National Key New Drug Creation Project, No. 2017ZX09304002.
Institutional review board statement: The study was reviewed and approved by the IRB of Shuguang Hospital affiliated with the Shanghai University of TCM Institutional Review Board.
Informed consent statement: All study participants, or their legal guardian, provided informed written consent prior to study enrollment.
Conflict-of-interest statement: There are no conflicts of interest to report.
STROBE statement: The authors have read the STROBE Statement checklist of items, and the manuscript was prepared and revised according to the STROBE Statement checklist of items.
Open-Access: This article is an open-access article which was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See: http://creativecommons.org/licenses/by-nc/4.0/
Correspondence to: Yi-Yang Hu, MD, PhD, Doctor, Senior Researcher, Institute of Liver Diseases, Key Laboratory of Liver and Kidney Diseases (Ministry of Education), Shuguang Hospital affiliated to Shanghai University of Traditional Chinese Medicine, No.528 Zhangheng Road, Pudong New Area, Shanghai 201203, China. yyhuliver@163.com
Telephone: +86-21-20256526 Fax: +86-21-20256521
Received: May 4, 2018
Peer-review started: May 4, 2018
First decision: May 23, 2018
Revised: June 8, 2018
Accepted: June 25, 2018
Article in press: June 25, 2018
Published online: August 14, 2018
Research background

Chronic hepatitis B is a major infectious disease in China and Chinese medicine has a wide range of applications in the treatment of CHB. Tongue diagnosis has important guiding significance for clinical syndrome differentiation and drug use in TCM, but lacks scientific explanations. Some reports have found abnormalities in the microbiota or metabolites in pathological tongue coatings. However, integrated analyses of the pathological tongue coating microbiotas and metabolites have rarely been reported. Elucidating the tongue coatings micro-features differences will promote our understanding of the TCM tongue diagnosis and facilitate therapeutic strategies for individualized treatment.

Research motivation

The motivation of this study was to explore the microfeatures of different tongue coatings, which could promote our understanding of the TCM tongue diagnosis from a modern perspective.

Research objectives

The objective of this research was to elucidate tongue coating microbiota and metabolic differences in CHB patients with yellow or white tongue coatings.

Research methods

We collected tongue coating samples from 28 CHB yellow tongue coating patients and 25 CHB white tongue coating patients, and an additional 22 samples were collected from healthy controls. The tongue coating bacterial 16S ribosomal RNA gene V3 region was amplified and sequenced with the Ion Torrent PGM™ sequencing platform. The metabolites in the tongue coatings were examined using a LC-MS platform. The microbiota results were analyzed using Metastats analysis, the Kruskal-Wallis test and LEfSe analysis. The functionality of the microbiota was assessed using PiCRUST and compared among groups. The metabolomics enrichment analysis was performed with MetaboAnalyst 3.0 (http://www.metaboanalyst.ca). We correlated the microbiota with the clinical indices and metabolites and visualized the results with Spearman’s correlation coefficients > 0.3 and P < 0.05 in Cytoscape.

Research results

This study found taxonomic and predicted function differences between the CHB yellow and white tongue coating patients. Distinct from those of the white coating patients, the CHB yellow tongue coating patients had specific clinical and microbiota characteristics. The microbiota of the CHB patients with yellow tongue coatings had similarities with a gut disorder in patients with cirrhosis at the phylum level, which manifested as a reduced abundance of Bacteroidetes and increased Proteobacteria content. Neisseriaceae, which is a dominant bacterial family enriched in yellow tongue coating patients, was positively correlated with the serum HBV-DNA level. The inferred metagenomic pathways enriched in the CHB yellow tongue coating patients were mainly those involved in amino acid metabolism, whereas the detected metabolites were mainly essential amino acids and generally were present in larger amounts than in the white tongue coatings.

Research conclusions

We found that the yellow and white tongue coatings of CHB patients had microbiota compositional and functional differences in this study.

Research perspectives

The sample size in this study was small. In the future, we will expand the sample size to further verify the results and focus on the relationship between changes in the oral and intestinal microbiotas to explore the effects of oral microbiota variance on immunity and metabolic alterations in the body. These studies will further explain the modern theoretical mechanism of TCM tongue diagnosis.