Basic Study
Copyright ©The Author(s) 2017. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. Jan 7, 2017; 23(1): 93-102
Published online Jan 7, 2017. doi: 10.3748/wjg.v23.i1.93
Role of mast cell-miR-490-5p in irritable bowel syndrome
Hai-Xia Ren, Fa-Can Zhang, He-Sheng Luo, Guo Zhang, Lie-Xin Liang
Hai-Xia Ren, He-Sheng Luo, Department of Gastroenterology, Renmin Hospital of Wuhan University, Wuhan 430060, Hubei Province, China
Fa-Can Zhang, Guo Zhang, Lie-Xin Liang, Department of Gastroenterology, Renmin Hospital of Guangxi Zhuang Autonomous Region, Nanning 530021, Guangxi Zhuang Autonomous Region, China
Author contributions: Zhang FC, Luo HS, Zhang G and Liang LX designed this research; Ren HX performed the research; Ren HX, Zhang FC and Luo HS analyzed the data; Ren HX wrote the paper.
Supported by the National Natural Science Foundation of China, No. 81160053 (to Zhang FC).
Institutional review board statement: The study was reviewed and approved by the People’s Hospital of Wuhan University and People’s Hospital of Guangxi Zhuang Autonomous Region Institutional Review Board.
Conflict-of-interest statement: We declare that we have no conflict of interest.
Data sharing statement: No additional data are available.
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: He-Sheng Luo, MD, Professor, Department of Gastroenterology, Renmin Hospital of Wuhan University, No. 238, Jiefang Road, Wuchang District, Wuhan 430060, Hubei Province, China. xhnk@163.com
Telephone: +86-27-97718831 Fax: +86-27-97718831
Received: July 13, 2016
Peer-review started: July 17, 2016
First decision: August 19, 2016
Revised: September 5, 2016
Accepted: September 14, 2016
Article in press: September 14, 2016
Published online: January 7, 2017
Processing time: 175 Days and 19.5 Hours
Abstract
AIM

To determine the functional role of miR-490-5p in mast cell proliferation and apoptosis, and in the mast cell tryptase/PAR-2 signal pathway.

METHODS

The 3rd generation of lentivirus vector systems containing enhanced green fluorescent protein (EGFP) (Ruisai Inc., Shanghai, China), which acts as a reporter gene was used to construct the mmu-miR-490-5p lentivirus expression vector pEGFP-antagomiR-490-5p, and the lentivirus vector pEGFP-negative was used as a negative control. The stably transfected mast cell line p815 was then constructed. GFP positive cells were successfully transfected cells. We determined the expression of miR-490-5p in p815 mast cells before and after transfection using quantitative real-time PCR (qRT-PCR). In addition, after transduction with the lentivirus vectors, the role of miR-490-5p in mast cell proliferation and apoptosis was investigated using the CCK-8 assay and flow cytometry, respectively. The mRNA levels of tryptase and PAR-2 were detected by qRT-PCR and the protein levels were detected by Western blot.

RESULTS

The inhibition of miR-490-5p expression promoted apoptosis and inhibited proliferation of p815 mast cells. The mRNA levels of tryptase and PAR-2 were significantly increased after transfection compared with the control group, tryptase (P = 0.721, normal vs null; P = 0.001, siRNA vs normal; P = 0.002, siRNA vs null) and PAR-2 (P = 0.027, siRNA vs null; P = 0.353, normal vs null; P = 0.105, siRNA vs normal). The protein levels of tryptase and PAR2 were slightly higher in the siRNA group than those in the control group, but the difference was not statistically significant (P > 0.05).

CONCLUSION

miR-490-5p plays a vital role in the pathogenesis of irritable bowel syndrome by affecting mast cell proliferation and apoptosis; with down-regulation of miR-490-5p, the mRNA level of mast cell tryptase and PAR-2 increased, and the protein level increased, but the difference was not statistically significant.

Keywords: miR-490-5p; Mast cell tryptase; PAR-2; Irritable bowel syndrome

Core tip: Mast cells and mast cell degranulation play important roles in the pathophysiology of irritable bowel syndrome. miRNA is a class of important endogenous single-stranded non-coding RNA, and plays an essential regulatory role in complex biological systems without protein translation. In the present study, we aimed to determine the role of miR-490-5p in regulating mast cell proliferation and apoptosis, the expression of mast cell tryptase and PAR-2, and thus predict the role of miR-490-5p in the pathogenesis of irritable bowel syndrome.