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Updated: Mar 8, 2026
Perturbations of Circulating miRNAs in Irritable Bowel Syndrome Detected Using a Multiplexed High-throughput Gene Expression Platform
Published on: November 30, 2016
Role of mast cell-miR-490-5p in irritable bowel syndrome
Hai-Xia Ren1, Fa-Can Zhang1, He-Sheng Luo1
1Hai-Xia Ren, He-Sheng Luo, Department of Gastroenterology, Renmin Hospital of Wuhan University, Wuhan 430060, Hubei Province, China.
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.
Insights
Inhibition of microRNA-490-5p (miR-490-5p) in mast cells promotes apoptosis and reduces proliferation. Down-regulation of miR-490-5p increases mRNA levels of mast cell tryptase and PAR-2.
Area of Science:
- Molecular Biology
- Cell Biology
- Immunology
Background:
- Mast cells play a crucial role in allergic inflammation and immune responses.
- MicroRNAs (miRNAs) are small non-coding RNAs that regulate gene expression and are implicated in various cellular processes.
- The specific role of miR-490-5p in mast cell function and its involvement in signaling pathways like tryptase/PAR-2 remains to be fully elucidated.
Purpose of the Study:
- To investigate the functional role of miR-490-5p in regulating mast cell proliferation and apoptosis.
- To determine the impact of miR-490-5p on the mast cell tryptase/PAR-2 signaling pathway.
Main Methods:
- Construction of a lentivirus expression vector for miR-490-5p inhibition in the p815 mast cell line.
- Quantitative real-time PCR (qRT-PCR) to measure miR-490-5p expression.
- CCK-8 assay and flow cytometry to assess mast cell proliferation and apoptosis.
- qRT-PCR and Western blot to analyze the mRNA and protein levels of tryptase and PAR-2.
Main Results:
- Inhibition of miR-490-5p significantly promoted apoptosis and inhibited proliferation of p815 mast cells.
- Down-regulation of miR-490-5p led to a significant increase in the mRNA levels of both tryptase and PAR-2.
- While protein levels of tryptase and PAR-2 showed a slight increase, the difference was not statistically significant.
Conclusions:
- miR-490-5p plays a significant role in mast cell proliferation and apoptosis.
- Down-regulation of miR-490-5p influences the expression of mast cell tryptase and PAR-2, suggesting its involvement in the tryptase/PAR-2 signaling pathway.
- These findings suggest miR-490-5p may be a potential therapeutic target in conditions involving mast cell dysregulation, such as irritable bowel syndrome.
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