Basic Study
Copyright ©The Author(s) 2019. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. May 7, 2019; 25(17): 2071-2085
Published online May 7, 2019. doi: 10.3748/wjg.v25.i17.2071
Herbs-partitioned moxibustion alleviates aberrant intestinal epithelial cell apoptosis by upregulating A20 expression in a mouse model of Crohn’s disease
Jing Zhou, Lu-Yi Wu, Liu Chen, Ya-Jing Guo, Yi Sun, Tao Li, Ji-Meng Zhao, Chun-Hui Bao, Huan-Gan Wu, Yin Shi
Jing Zhou, Liu Chen, Ya-Jing Guo, Yi Sun, Tao Li, Graduate School, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China
Lu-Yi Wu, Qigong Institute, Shanghai University of Traditional Chinese Medicine, Shanghai 200030, China
Ji-Meng Zhao, Chun-Hui Bao, Huan-Gan Wu, Yin Shi, Key Laboratory of Acupuncture and Immunological Effects, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China
Ji-Meng Zhao, Chun-Hui Bao, Huan-Gan Wu, Key Laboratory of Acupuncture and Immunological Effects, Shanghai Institute of Acupuncture-Moxibustion and Meridian, Shanghai 200030, China
Yin Shi, Outpatient Department, Shanghai Institute of Acupuncture-Moxibustion and Meridian, Shanghai 200030, China
Author contributions: Zhou J, Shi Y, and Wu HG designed the research; Zhou J, Chen L, Zhao JM, Guo YJ, Sun Y, and Li T performed the experiments; Wu LY, Zhou J, and Bao CH collected and analyzed the data; Zhou J wrote the manuscript; all authors reviewed the manuscript prior to its submission, and read and approved the final manuscript.
Supported by National Natural Science Foundation of China, No. 81273844 and No. 81473757; National Key Basic Research Program of China (973 Program), No. 2015CB554500; and Shanghai Rising-Star Program, No. 16QA1403400.
Institutional animal care and use committee statement: All animal experiments in this study were performed under guidelines approved by the Animal Ethics Committee of the Shanghai University of Traditional Chinese Medicine (No. 2013025).
Conflict-of-interest statement: The authors declare no conflicts of interest.
Data sharing statement: No additional data are available.
ARRIVE guidelines statement: The manuscript was prepared according to the ARRIVE guidelines.
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/
Corresponding author: Yin Shi, PhD, Doctor, Professor, Outpatient Department, Shanghai Institute of Acupuncture-Moxibustion and Meridian, 650 South Wanping Road, Shanghai 200030, China. flysy0636@163.com
Telephone: +86-21-64383910 Fax: +86-21-64390339
Received: January 13, 2019
Peer-review started: January 14, 2019
First decision: February 13, 2019
Revised: March 13, 2019
Accepted: March 15, 2019
Article in press: March 16, 2019
Published online: May 7, 2019
Abstract
BACKGROUND

A20 inhibits intestinal epithelial cell apoptosis in Crohn’s disease, and herbs-partitioned moxibustion (HPM) has been demonstrated to be an effective treatment for Crohn’s disease. However, the mechanism by which HPM reduces intestinal epithelial cell apoptosis in Crohn’s disease has not been thoroughly elucidated to date.

AIM

To elucidate whether HPM exerts its effects by upregulating A20 to affect intestinal epithelial cell apoptosis in a Crohn’s disease mouse model.

METHODS

In this study, mice with A20 deletion in intestinal epithelial cells (A20IEC-KO) were utilized to establish a Crohn’s disease mouse model with 2,4,6-trinitrobenzene sulfonic acid (TNBS) administration, as well as wild-type mice. Mice were randomly divided into normal control (NC), model control (MC), mesalazine (MESA), and HPM groups. The morphology of the colonic mucosa was observed by hematoxylin-eosin staining, and serum endotoxin and apoptosis of epithelial cells were evaluated by enzyme-linked immunosorbent assay and terminal dUTP nick-end labeling assay accordingly. The protein expression levels of A20 and tumor necrosis factor receptor 1 (TNFR1)-related signaling molecules were evaluated by Western blot, and co-expression of A20 and TNFR1-associated death domain (TRADD) and co-expression of A20 and receptor-interacting protein 1 (RIP1) were observed by double immunofluorescence staining.

RESULTS

The intestinal epithelial barrier was noted to have an improvement in the HPM group of wild-type (WT) mice compared with that in A20IEC-KO mice. Compared with A20 IEC-KO HPM mice, serum endotoxin levels and apoptosis percentages were decreased (P < 0.01), A20 expression levels were increased (P < 0.01), and expression of TNFR1, TRADDD, and RIP1 was decreased in the HPM group of WT mice (PTNFR1 < 0.05, PTRADD < 0.01, PRIP1 < 0.01). Both of the co-expression of A20/TRADD and A20/RIP1 showed a predominantly yellow fluorescence in the HPM group of WT mice, while a predominantly red fluorescence was noted in the HPM group of A20IEC-KO mice.

CONCLUSION

Our findings suggest that HPM in treating Crohn’s disease functions possibly via upregulation of the A20 expression level, resulting in downregulation of TNFR1, TRADD, and RIP1 to alleviate increased cell apoptosis in the intestinal epithelial barrier in Crohn's disease.

Keywords: Herbs-partitioned moxibustion, Crohn’s disease, Apoptotic pathway, Inflammation, A20

Core tip: We report our results derived from mice with A20 deletion in intestinal epithelial cells by inducing Crohn’s disease. The Crohn’s disease model was induced with 2,4,6-trinitrobenzene sulfonic acid. This study demonstrates for the first time that herbs-partitioned moxibustion can upregulate the expression of A20, resulting in downregulation of tumor necrosis factor receptor (TNFR) 1, TNFR1-associated death domain, and receptor-interacting protein 1 to alleviate increased cell apoptosis in the intestinal epithelial barrier in Crohn's disease.