Basic Study
Copyright ©The Author(s) 2019. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. Sep 14, 2019; 25(34): 5134-5151
Published online Sep 14, 2019. doi: 10.3748/wjg.v25.i34.5134
Effect of Tong Xie Yao Fang on endogenous metabolites in urine of irritable bowel syndrome model rats
Xue-Ying Zhao, Jian-Wei Wang, Yue Yin, Kai Li, Miao Zhang, Fu-Ping Yan
Xue-Ying Zhao, Jian-Wei Wang, Yue Yin, Kai Li, Miao Zhang, Fu-Ping Yan, School of Basic Medical Sciences, Heilongjiang University of Chinese Medicine, Harbin 150040, Heilongjiang Province, China
Author contributions: Zhao XY and Wang JW performed most of the experiments, analyzed the experimental results, interpreted them and wrote the manuscript; Yin Y, Li K, Zhang M and Yan FP performed some experiments and interpretation of the results; Wang JW participated in the testing factor design and revised the manuscript. She is also the corresponding author; All co-authors participated in writing and checking the manuscript and approved the submitted manuscript.
Supported by the National Natural Science Foundation of China, No. 81573870; the Eighth Special Subsidy Project of China Post-Doctoral Science Foundation, No. 2015T80376; Postdoctoral Science-Research Developmental Foundation of China, No. 2013M531079; National Science Foundation of Heilongjiang Province, No. H2015020; Heilongjiang University of Traditional Chinese Medicine Outstanding Innovative Talents Support Project (Outstanding Young Academic Leaders), Postdoctoral Science-Research Developmental Foundation of Heilongjiang Province, No. LBH-Q12009; Youth Academic Backbone Fund of Heilongjiang Province Education Department, No. 1253G053; Youth Science and Technology Project of Traditional Chinese Medicine Department of Heilongjiang Province, No. ZQG-034.
Institutional review board statement: This study was approved by the Ethics Committee of the Heilongjiang University of Chinese Medicine.
Institutional animal care and use committee statement: This study was approved by the animal care and use committee of the Heilongjiang University of Chinese Medicine.
Conflict-of-interest statement: Authors declare no conflicts of interest.
Data sharing statement: The detailed experimental methods are available from the corresponding author at xueyingzhao2010@126.com and wangjianwei140918@126.com.
ARRIVE guidelines statement: The ARRIVE Guidelines have been adopted.
Open-Access: This is an open-access article that 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/
Corresponding author: Jian-Wei Wang, PhD, Professor, School of Basic Medical Sciences, Heilongjiang University of Chinese Medicine, Heping Road 24, Harbin 150040, Heilongjiang Province, China. wangjianwei140918@126.com
Telephone: +86-451-82193640
Received: April 18, 2019
Peer-review started: April 18, 2019
First decision: July 22, 2019
Revised: July 30, 2019
Accepted: August 19, 2019
Article in press: August 19, 2019
Published online: September 14, 2019
ARTICLE HIGHLIGHTS
Research background

The establishment of disease and syndrome combined with animal model in traditional Chinese medicine (TCM) is an important link to realize the objectiveness, standardization and scientificity of TCM. At present, the model construction method of the disease and syndrome combination for irritable bowel syndrome (IBS) is mainly based on the modern medical disease animal model combined with syndrome related etiology and disease mechanism. There are obvious differences in metabolic products among TCM syndromes. Metabonomics finds biomarkers of diseases by comparing the metabolic profiles of animal body fluids, analyzing the differences of metabolic pathways and understanding the pathological process and metabolic pathways of substances in the body. Thus, the evaluation of animal models tends to be comprehensive and objective. Therefore, through the combination of metabonomics and other systems biology with TCM prescription syndrome, we can clearly define the attributes of animal model syndrome. Through identical formulas, different models and different formulas, we can determine the model of syndrome from the perspective of metabolic products in vivo, clarify the mechanism and essence of prescriptions and drugs and clarify the correlation between prescriptions and syndromes. This research idea not only conforms to the research requirements of TCM prescriptions and syndromes but also follows the regularity between phenomena and essence in modern science.

Research motivation

Tong Xie Yao Fang is effective in the treatment of IBS. The overall concept of TCM theory and the treatment of evidence differentiation require that the clinical differentiation of IBS must be accurate. Therefore, the establishment of a syndrome-binding IBS model is the key to research of IBS in TCM. In recent years, the study of disease syndrome combined with animal model has developed rapidly and has become a new direction of animal model development in TCM research. At present, many scholars have provided a suitable combination model of disease and syndrome for the study of TCM in the treatment of IBS, which reflects the advantages of TCM on IBS syndrome differentiation. Therefore, in the aspect of disease and syndrome combination model replication, it is closely related to the advanced research methods of modern medicine. Through the combination of metabonomics and other system biology with TCM prescription syndrome, the attributes of IBS disease syndrome combined with animal model syndrome can be clearly defined. Through the improvement of the construction method and train of thought of the disease syndrome model, a more mature disease syndrome combined with animal model would be obtained, and the scientific connotation of TCM will be expounded.

Research objectives

The research objective is to discover the biomarkers of Tong Xie Yao Fang and establish its metabolic model characteristics through its influence on metabolites of IBS. The action essence of Tong Xie Yao Fang and the syndrome essence of its disease model are explored based on the changing law of substance quality in vivo.

Research methods

In this study, Wistar rats were used to establish the IBS models, and then randomly divided into four groups: Model control group, Tong Xie Yao Fang treatment groups (high, medium, low doses). A normal, non-IBS group was established. The rats were treated for 2 wk. On days 0 and 14 of the experimental model, urine was collected for 12 h and was analyzed by ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry. Nine potential biomarkers were identified, and six major metabolic pathways were found to be related to IBS.

Research results

In the study of metabonomics, nine potential biomarkers including L-serine, 4-methylgallic acid, L-threonine, succinylacetone, prolyl-hydroxyproline, valyl-serine, acetyl citrate, marmesin rutinoside and 5-hydroxy-L-tryptophan were identified in urine, which were assigned to amino acids, organic acids, succinyl and glycosides. Furthermore, the metabolic pathway of L-serine, L-threonine and 5-hydroxy-L-tryptophan was found in the Kyoto Encyclopedia of Genes and Genomes, which mainly involved the metabolism of cysteine and methionine, vitamin B6 metabolism, serotonin synapse, tryptophan metabolism, sphingolipid metabolism, digestion and absorption of protein and amino acid metabolism. These pathways are related to intestinal dysfunction, inflammatory syndrome, nervous system dysfunction and other diseases. However, due to the complexity of regulation, we do not fully understand the biological significance of abnormal expression of endogenous metabolites.

Research conclusions

The endogenous metabolites changed significantly for the IBS model after the intervention of Tong Xie Yao Fang. Tong Xie Yao Fang plays a complex regulatory role in endogenous metabolites associated with liver spleen deficiency syndrome. It is suggested that the improvement of spleen deficiency and liver hyperactivity of IBS by Tong Xie Yao Fang is related to the regulation of the serotonin synaptic pathway and tryptophan metabolism. It was confirmed that the rectal dilatation stimulation combined with maternal-infant separation of IBS animal model was the best model for the adaptation of Tong Xie Yao Fang. In this study, through metabonomics related research on Tong Xie Yao Fang, endogenous metabolites in animal models were evident, suggesting the importance of syndrome differentiation and treatment of clinical syndrome in TCM.

Research perspectives

Metabonomics, genomics, transcriptome, and proteomics have the characteristics of systematic methodology such as overall dynamics, synthesis and analysis and are similar to the overall concept of TCM in the treatment of diseases. By means of systematic biology research method, prescriptions and syndromes can be integrated into an organic system for research. Therefore, the subject not only meets the research requirements of prescriptions and syndromes in TCM but also conforms to the regularity of the relationship between phenomena and essence in modern science.