Basic Study
Copyright ©The Author(s) 2023. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastrointest Surg. Apr 27, 2023; 15(4): 600-620
Published online Apr 27, 2023. doi: 10.4240/wjgs.v15.i4.600
Acinous cell AR42J-derived exosome miR125b-5p promotes acute pancreatitis exacerbation by inhibiting M2 macrophage polarization via PI3K/AKT signaling pathway
Zhi Zheng, Feng Cao, Yi-Xuan Ding, Jiong-Di Lu, Yuan-Qiao Fu, Lin Liu, Yu-Lin Guo, Shuang Liu, Hai-Chen Sun, Ye-Qing Cui, Fei Li
Zhi Zheng, Yuan-Qiao Fu, Department of General Surgery, Beijing Friendship Hospital, Capital Medical University, Beijing 100050, China
Feng Cao, Yi-Xuan Ding, Jiong-Di Lu, Yu-Lin Guo, Shuang Liu, Hai-Chen Sun, Ye-Qing Cui, Fei Li, Department of General Surgery, Xuanwu Hospital, Capital Medical University, Beijing 100053, China
Lin Liu, Department of Gastroenterology, The Second Affiliated Hospital of Baotou Medical College, Baotou 014040, Inner Mongolia Autonomous Region, China
Author contributions: Zheng Z, Cao F, Ding YX, Lu JD, Fu YQ and Liu L are equally contributed to this work; Zheng Z, Ding YX and Lu JD designed the study; Liu L, Guo YL, Liu S, Sun HC and Cui YQ performed the experiments; Zheng Z and Lu JD wrote the manuscript; Sun HC, Fu YQ and Cui YQ performed statistical analysis; Li F and Cao F revised the manuscript; All authors read and approved the final manuscript.
Supported by The Beijing Municipal Science and Technology Commission, Capital Research and Demonstration Application of Clinical Diagnosis and Treatment Technology, No. Z191100006619038 and No. Z171100001017077; and The Capital Health Research and Development of Special, No. 2020-1-2012.
Institutional review board statement: The research was given the go-ahead by the hospital’s Ethics Committee and Animal Ethics Committee, which is in Xuanwu Hospital, Capital Medical University, No. 2020-158.
Institutional animal care and use committee statement: All experimental procedures and feeding management methods involving animals have been reviewed and approved by the Animal Experiment Ethics Committee of Xuanwu Hospital of Capital Medical University, No. 2020-158.
Informed consent statement: All study participants or their legal guardian provided informed written consent about personal and medical data collection prior to study enrolment.
Conflict-of-interest statement: All the authors report no relevant conflicts of interest for this article.
Data sharing statement: Data are available from the corresponding authors upon request.
ARRIVE guidelines statement: The authors have read the ARRIVE guidelines, and the manuscript was 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: Fei Li, MD, PhD, Deputy Director, Professor, Surgeon, Department of General Surgery, Xuanwu Hospital, Capital Medical University, No. 45 Chang-chun Street, Xi-Cheng District, Beijing 100053, China. feili36@ccmu.edu.cn
Received: October 30, 2022
Peer-review started: October 30, 2022
First decision: January 3, 2023
Revised: January 5, 2023
Accepted: March 8, 2023
Article in press: March 8, 2023
Published online: April 27, 2023
Processing time: 175 Days and 4.6 Hours
Abstract
BACKGROUND

The incidence rate of acute pancreatitis (AP), which is a pathophysiological process with complex etiology, is increasing globally. miR-125b-5p, a bidirectional regulatory miRNA, is speculated to exhibit anti-tumor activity. However, exosome-derived miR-125b-5p in AP has not been reported.

AIM

To elucidate the molecular mechanism of exosome-derived miR-125b-5p promoting AP exacerbation from the perspective of the interaction between immune cells and acinar cells.

METHODS

Exosomes derived from AR42J cells were isolated and extracted in active and inactive states by an exosome extraction kit, and were verified via transmission electron microscopy, nanoparticle tracking analysis, and western blotting. RNA sequencing assay technology was used to screen differentially expressed miRNAs in active and inactive AR42J cell lines, and bioinformatics analysis was used to predict downstream target genes of miR-125b-5p. The expression level of miR-125b-5p and insulin-like growth factor 2 (IGF2) in the activated AR42J cell line and AP pancreatic tissue were detected by quantitative real-time polymerase chain reaction and western blots. The changes in the pancreatic inflammatory response in a rat AP model were detected by histopathological methods. Western Blot was used to detect the expression of IGF2, PI3K/AKT signaling pathway proteins, and apoptosis and necrosis related proteins.

RESULTS

miR-125b-5p expression was upregulated in the activated AR42J cell line and AP pancreatic tissue, while that of IGF2 was downregulated. In vitro experiments confirmed that miR-125b-5p could promote the death of activated AR42J cells by inducing cell cycle arrest and apoptosis. In addition, miR-125b-5p was found to act on macrophages to promote M1 type polarization and inhibit M2 type polarization, resulting in a massive release of inflammatory factors and reactive oxygen species accumulation. Further research found that miR-125b-5p could inhibit the expression of IGF2 in the PI3K/AKT signaling pathway. Additionally, in vivo experiments revealed that miR-125b-5p can promote the progression of AP in a rat model.

CONCLUSION

miR-125b-5p acts on IGF2 in the PI3K/AKT signaling pathway and promotes M1 type polarization and inhibits M2 type polarization of macrophage by inhibiting IGF2 expression, resulting in a large release of pro-inflammatory factors and an inflammatory cascade amplification effect, thus aggravating AP.

Keywords: Acute pancreatitis; Exosome; miR-125b-5p; Macrophage; Mechanism

Core Tip: Our study confirmed that miR-125b-5p can promote the inflammatory response of acute pancreatitis (AP), and its potential targets were found via RNA sequencing assay. However, the likely mechanism remains unclear. Consequently, our study intends to elucidate the molecular mechanism of exosome-derived miR-125b-5p in promoting AP exacerbation from the perspective of the interaction between immune cells and acinar cells.