Basic Study
Copyright ©The Author(s) 2022. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. Jul 28, 2022; 28(28): 3644-3665
Published online Jul 28, 2022. doi: 10.3748/wjg.v28.i28.3644
Mechanism of electroacupuncture and herb-partitioned moxibustion on ulcerative colitis animal model: A study based on proteomics
Qin Qi, Rui Zhong, Ya-Nan Liu, Chen Zhao, Yan Huang, Yuan Lu, Zhe Ma, Han-Dan Zheng, Lu-Yi Wu
Qin Qi, Ya-Nan Liu, Yan Huang, Yuan Lu, Zhe Ma, Han-Dan Zheng, Lu-Yi Wu, Yueyang Hospital of Integrated Traditional Chinese and Western Medicine, Shanghai University of Traditional Chinese Medicine, Shanghai 200437, China
Rui Zhong, Shanghai QiGong Research Institute, Shanghai University of Traditional Chinese Medicine, Shanghai 200030, China
Chen Zhao, School of Acupuncture, Moxibustion and Tuina, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China
Author contributions: Wu LY, Huang Y and Qi Q conceived and designed this study; Zhong R, Liu YN, Ma Z and Zheng HD performed the animal experiments, acquired and analyzed the data; Qi Q wrote the main manuscript; Liu YN and Ma Z prepared the figures and tables; Lu Y gave guidance to the manuscript writing; Zhao C, Huang Y and Wu LY revised and improved the manuscript; and all authors reviewed and approved the final version of this manuscript.
Supported by the National Natural Science Foundation of China No. 81973955, 82004475 and 82174501; Shanghai Clinical Research Center for Acupuncture and Moxibustion No. 20MC1920500; Clinical Key Specialty Construction Foundation of Shanghai No. shslczdzk04701; Natural Science Foundation of Shanghai No. 21ZR1460200.
Institutional animal care and use committee statement: All animal experiments were performed according to the protocols approved by the Animal Ethics Committee of the Experimental Animal Center of Shanghai University of Traditional Chinese Medicine.
Conflict-of-interest statement: All the authors report no relevant conflicts of interest for this article.
Data sharing statement: No additional data are available.
ARRIVE guidelines statement: The authors have read the ARRIVE guidelines, and the manuscript has been prepared and revised according to the ARRIVE guidelines.
Open-Access: This article 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 NonCommercial (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: https://creativecommons.org/Licenses/by-nc/4.0/
Corresponding author: Lu-Yi Wu, PhD, Associate Professor, Yueyang Hospital of Integrated Traditional Chinese and Western Medicine, Shanghai University of Traditional Chinese Medicine, No. 110 Ganhe Road, Hongkou District, Shanghai 200437, China.luyitcm@163.com
Received: September 3, 2021
Peer-review started: September 3, 2021
First decision: November 7, 2021
Revised: November 19, 2021
Accepted: June 24, 2022
Article in press: June 24, 2022
Published online: July 28, 2022
Processing time: 326 Days and 23.4 Hours
Abstract
BACKGROUND

Ulcerative colitis (UC) is a chronic, nonspecific intestinal inflammatory disease. Acupuncture and moxibustion is proved effective in treating UC, but the mechanism has not been clarified. Proteomic technology has revealed a variety of biological markers related to immunity and inflammation in UC, which provide new insights and directions for the study of mechanism of acupuncture and moxibustion treatment of UC.

AIM

To investigate the mechanism of electroacupuncture (EA) and herb-partitioned moxibustion (HM) on UC rats by using proteomics technology.

METHODS

Male Sprague-Dawley rats were randomly divided into the normal (N) group, the dextran sulfate sodium (DSS)-induced UC model (M) group, the HM group, and the EA group. UC rat model was prepared with 3% DSS, and HM and EA interventions at the bilateral Tianshu and Qihai acupoints were performed in HM or EA group. Haematoxylin and eosin staining was used for morphological evaluation of colon tissues. Isotope-labeled relative and absolute quantification (iTRAQ) and liquid chromatography-tandem mass spectrometry were performed for proteome analysis of the colon tissues, followed by bioinformatics analysis and protein-protein interaction networks establishment of differentially expressed proteins (DEPs) between groups. Then western blot was used for verification of selected DEPs.

RESULTS

The macroscopic colon injury scores and histopathology scores in the HM and EA groups were significantly decreased compared to the rats in the M group (P < 0.01). Compared with the N group, a total of 202 DEPs were identified in the M group, including 111 up-regulated proteins and 91 down-regulated proteins, of which 25 and 15 proteins were reversed after HM and EA interventions, respectively. The DEPs were involved in various biological processes such as biological regulation, immune system progression and in multiple pathways including natural killer cell mediated cytotoxicity, intestinal immune network for immunoglobulin A (IgA) production, and FcγR-mediated phagocytosis. The Kyoto Encyclopedia of Genes and Genomes pathways of DEPs between HM and M groups, EA and M groups both included immune-associated and oxidative phosphorylation. Network analysis revealed that multiple pathways for the DEPs of each group were involved in protein-protein interactions, and the expression of oxidative phosphorylation pathway-related proteins, including ATP synthase subunit g (ATP5L), ATP synthase beta subunit precursor (Atp5f), cytochrome c oxidase subunit 4 isoform 1 (Cox4i1) were down-regulated after HM and EA interventions. Subsequent verification of selected DEPs (Synaptic vesicle glycoprotein 2A; nuclear cap binding protein subunit 1; carbamoyl phosphate synthetase 1; Cox4i1; ATP synthase subunit b, Atp5f1; doublecortin like kinase 3) by western blot confirmed the reliability of the iTRAQ data, HM and EA interventions can significantly down-regulate the expression of oxidative phosphorylation-associated proteins (Cox4i1, Atp5f1) (P < 0.01).

CONCLUSION

EA and HM could regulate the expression of ATP5L, Atp5f1, Cox4i1 that associated with oxidative phosphorylation, then might regulate immune-related pathways of intestinal immune network for IgA production, FcγR-mediated phagocytosis, thereby alleviating colonic inflammation of DSS-induced UC rats.

Keywords: Proteomics, Ulcerative colitis, Moxibustion, Electroacupuncture, Differential proteins

Core Tip: Ulcerative colitis (UC) is a nonspecific inflammatory bowel disease with unclear etiology. Acupuncture and moxibustion are benefit to UC by improving colonic mucosa damage, regulating inflammatory cytokines. In recent years, proteomic technology has been widely used in the study of UC, revealing a variety of biological markers related to immunity and inflammation in UC. We applied isotope-labeled relative and absolute quantification proteomics technology to identify UC-relevant protein targets and further explore the mechanism of acupuncture and moxibustion. It was found that electroacupuncture and herb-partitioned moxibustion could regulate the expression of multiple proteins, such as ATP synthase subunit g, ATP synthase beta subunit precursor 1, cytochrome c oxidase subunit 4 isoform 1 that associated with oxidative phosphorylation, that might regulate immune-related pathways, thereby alleviating colonic inflammation of dextran sulfate sodium-induced UC rats.