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Detection of microRNA Expression in Peritoneal Membrane of Rats Using Quantitative Real-time PCR
Published on: June 27, 2017
MicroRNA and mRNA expression profiling in rat acute respiratory distress syndrome
Chaoqun Huang, Xiao Xiao, Narendranath Reddy Chintagari
1Department of Physiological Sciences, Lundberg-Kienlen Lung Biology and Toxicology Laboratory, Stillwater, OK, USA. lin.liu@okstate.edu.
Background:
Acute respiratory distress syndrome (ARDS) is characterized by pulmonary epithelial injury and extensive inflammation of the pulmonary parenchyma. Systematic analyses of microRNA (miRNA) and mRNA expression profiling in ARDS provide insights into understanding of molecular mechanisms of the pathogenesis of ARDS. The objective of this study was to identify miRNA and mRNA interactions in a rat model of ARDS by combining miRNA and mRNA microarray analyses.
Methods:
Rat model of ARDS was induced by saline lavage and mechanical ventilation. The expression profiles of both mRNAs and miRNAs in rat ARDS model were performed by microarray analyses. Microarray data were further verified by quantitative RT-PCR. Functional annotation on dys-regulated mRNAs and miRNAs was carried out by bioinformatics analysis.
Results:
The expression of 27 miRNAs and 37 mRNAs were found to be significantly changed. The selected miRNAs and genes were further verified by quantitative real-time PCR. The down-regulated miRNAs included miR-24, miR-26a, miR-126, and Let-7a, b, c, f. The up-regulated miRNAs were composed of miR-344, miR-346, miR-99a, miR-127, miR-128b, miR-135b, and miR-30a/b. Gene ontology and functional annotation analyses indicated that up-regulated mRNAs, such as Apc, Timp1, and Sod2, were involved in the regulation of apoptosis. Bioinformatics analysis showed the inverse correlation of altered miRNAs with the expression of their predicted target mRNAs. While Sod2 was inversely correlated with Let-7a, b, c, f., Ebf1 and Apc were inversely correlated with miR-24 and miR-26a, respectively. miR-26a, miR-346, miR-135b, miR-30a/b, miR-344, and miR-18a targeted multiple altered mRNAs. Gabrb1, Sod2, Eif2ak1, Fbln5, and Tspan8 were targeted by multiple altered miRNAs.
Conclusion:
The expressions of miRNAs and mRNAs were altered in a rat model of ARDS. The identified miRNA-mRNA pairs may play critical roles in the pathogenesis of ARDS.
Insights
This study identified altered microRNA (miRNA) and messenger RNA (mRNA) expressions in a rat model of acute respiratory distress syndrome (ARDS). These identified miRNA-mRNA interactions may be crucial in ARDS development.
Area of Science:
- Pulmonary Medicine
- Molecular Biology
- Genomics
Background:
- Acute respiratory distress syndrome (ARDS) involves lung injury and inflammation.
- Understanding the molecular mechanisms of ARDS pathogenesis is crucial.
- MicroRNA (miRNA) and mRNA expression profiling offers insights into ARDS pathology.
Purpose of the Study:
- To identify miRNA and mRNA interactions in a rat model of ARDS.
- To combine miRNA and mRNA microarray analyses for comprehensive profiling.
- To elucidate molecular mechanisms underlying ARDS pathogenesis.
Main Methods:
- Induction of ARDS in a rat model using saline lavage and mechanical ventilation.
- Microarray analysis to determine miRNA and mRNA expression profiles.
- Quantitative RT-PCR for verification and bioinformatics for functional annotation.
Main Results:
- Significant changes observed in 27 miRNAs and 37 mRNAs.
- Down-regulated miRNAs include miR-24, miR-26a, miR-126, and Let-7 family; up-regulated miRNAs include miR-344, miR-346, miR-99a, and others.
- Bioinformatics analysis revealed inverse correlations between specific miRNAs and their target mRNAs, suggesting roles in apoptosis regulation.
Conclusions:
- miRNA and mRNA expressions are significantly altered in the ARDS rat model.
- Identified miRNA-mRNA pairs are potential key players in ARDS pathogenesis.
- Further research into these interactions could lead to novel therapeutic targets for ARDS.
