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Mature and immature microRNA ratios in cultured rat cardiomyocytes during anoxia-reoxygenation.

ABSTRACT: BACKGROUND:The critical role of microRNAs (miRNAs) in the global control of gene expression in the heart has recently been postulated; however, the mechanisms of miRNA regulation in cardiac pathology are not clear. OBJECTIVE:To evaluate the levels of miR-1, miR-208a and miR-29a expressed in neonatal rat cardiomyocytes during anoxia-reoxygenation (AR). METHODS:Reverse transcription coupled with real-time polymerase chain reaction was used to evaluate the level of mature and immature miRNAs in cardiomyocyte culture during AR. RESULTS:THE INITIAL LEVELS OF THE MATURE AND IMMATURE MIRNAS WERE DIFFERENT: mature - miR-1 7.46±4.440, miR-208a 0.02±0.015 and miR-29a 5.60±2.060; immature - miR-1 0.02±0.007, miR-208a 0.05±0.029 and miR-29a 0.01±0.008. The most prominent changes were observed for immature miRNAs during AR, with immature miR-1 and miR-29a expressed at significantly higher levels during remote reoxygenation (AR [0.5 h/24 h]) compared with control, while the level of expressed immature miR-208a was significantly decreased during acute reoxygenation (AR [0.5 h /1 h]) and returned to control levels during remote reoxygenation (AR [0.5h /24 h]). Also, the ratios of mature to immature miRNAs were significantly increased during acute reoxygenation for miR-1 and miR-208a, returning to control levels during remote reoxygenation, while for miR-29a, this ratio had the progressive tendency to decrease under AR. CONCLUSION:The discordance between the estimated levels of mature and immature miRNA during AR supports the hypothesis that transcriptional and post-transcriptional regulatory mechanisms at the miRNA level play a role in the response of cardiomyocytes to AR, and could be a contributing factor in the differential resistance of cardiomyocytes to AR.

SUBMITTER: Dosenko VE

PROVIDER: S-EPMC3395463 | biostudies-literature | 2012

REPOSITORIES: biostudies-literature

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Mature and immature microRNA ratios in cultured rat cardiomyocytes during anoxia-reoxygenation.

Dosenko Victor E VE Gurianova Veronika L VL Surova Olga V OV Stroy Dmitriy A DA Moibenko Alex A AA

Experimental and clinical cardiology 20120101 2

<h4>Background</h4>The critical role of microRNAs (miRNAs) in the global control of gene expression in the heart has recently been postulated; however, the mechanisms of miRNA regulation in cardiac pathology are not clear.<h4>Objective</h4>To evaluate the levels of miR-1, miR-208a and miR-29a expressed in neonatal rat cardiomyocytes during anoxia-reoxygenation (AR).<h4>Methods</h4>Reverse transcription coupled with real-time polymerase chain reaction was used to evaluate the level of mature and ...[more]

PMID: 22826655

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Project description:INTRODUCTION:The precise mechanisms of the inflammatory responses after cerebral ischemia in vivo are difficult to elucidate because of the complex nature of multiple series of interactions between cells and molecules. This study explored temporal patterns of secretion of 30 cytokines and chemokines from Sprague Dawley rat astrocytes in primary culture in order to elucidate signaling pathways that are triggered by astrocytes during anoxia. METHODS:Primary cultures of rat brain astrocytes were incubated for periods of 2-24 hr in the absence of oxygen (anoxia) or under normal partial pressure of oxygen (controls). Simultaneous detection of 29 cytokines and chemokines in the samples was performed using a rat cytokine array panel, while the temporal pattern of angiopoietin-1 (Ang-1) secretion was determined separately using ELISA. Wilcoxon-Mann-Whitney test was used to compare normoxic and anoxic samples and the Hodge-Lehman estimator with exact 95% confidence intervals was computed to assess the size of differences in cytokine secretion. The obtained data were imported into the Core Analysis tool of Ingenuity Pathways Analysis software in order to relate changes in secretion of cytokines and chemokines from astrocytes during anoxia to potential molecular signal networks. RESULTS:With the exception of Ang-1, concentrations of all cytokines/chemokines in samples collected after anoxia exposure were either the same, or higher, than in control groups. No clear pattern of changes could be established for groups of cytokines with similar effects (i.e., pro- or anti-inflammatory cytokines). The pattern of changes in cytokine secretion during anoxia was associated with the HIF-1?-mediated response, as well as cytokines IL-1? and cathepsin S pathways, which are related to initiation of inflammation and antigen presentation, respectively, and to ciliary neurotrophic factor. CONCLUSIONS:These in vitro findings suggest that astrocytes may play a role in triggering inflammation during anoxia/ischemia of the brain.

Dataset Information

Mature and immature microRNA ratios in cultured rat cardiomyocytes during anoxia-reoxygenation.

Publications

Mature and immature microRNA ratios in cultured rat cardiomyocytes during anoxia-reoxygenation.

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