Project description:We use the illumina high-throughput sequencing technology to identify miRNAs between the exosomes of H9c2 cells with or without alcohol-induced.The H9c2 cells were cultured in serum-free medium and stimulated with ethanol (100 mmol/L) or PBS for 24 h, then collected the exosomes samples from serum-free medium. Exosomes were isolated and extracted by differential centrifugation and detected by electron microscopy, particle size and related marker proteins.In total, 123 differentially expressed miRNAs (12 upregulated and 111 downregulated) were screened by miRNA sequence.
Project description:To investigate the function of exosomes from cardiopulmonary progenitors in the regulation of gene expression in endothelial cells.We performed gene expression profiling analysis using data obtained from RNA-seq of H9C2 treated with or without CPPs exosomes.
Project description:Chronic alcohol exposure can cause myocardial degenerative diseases, manifested as cardiac insufficiency, arrhythmia, etc. These are defined as alcoholic cardiomyopathy (ACM). Alcohol-mediated myocardial injury has previously been studied through metabolomics, and it has been proved to be involved in the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway related to the biosynthesis of unsaturated fatty acids and oxidative phosphorylation, which tentatively explored the mechanism of ACM induced by chronic drinking. To further study the myocardial damage caused by alcohol, the mouse model of ACM successfully established previously was used to perform proteomics analysis on myocardial specimens. Fifty-six differentially expressed proteins (DEPs) were identified, and they are involved in the KEGG pathway related to fatty acid biosynthesis, lipid metabolism, oxidative stress, and the development of dilated cardiomyopathy (DCM). The present study further demonstrated the underlying causes of myocardial damage caused by chronic alcohol consumption and lays the foundation for further research to clarify the underlying mechanism of ACM.
Project description:Ischemic heart disease continues to rank highly among cardiovascular diseases in the world. Myocardial reperfusion (R) is provided with an effective and rapid treatment, however it can lead to fatal results as well as ischemia (I). The goal of this study was to use proteomic analysis to assess the proteins and pathways that changed in H9C2 cardiomyocyte cells exposed to (I) and (R) for durations that represented acute and chronic conditions.
Project description:Aims: Mesenchymal stem cells (MSCs) gradually become attractive candidates for cardiac inflammation modulation, yet understanding of the mechanism remains elusive. Strikingly, recent studies indicated that exosomes secreted by MSCs might be a novel mechanism for the beneficial effect of MSCs transplantation after myocardial infarction. We therefore explored the role of MSC-derived exosomes (MSC-Exo) in the immunomodulation of macrophages after myocardial ischemia-reperfusion and its implications in cardiac injury repair. Methods and Results: Exosomes were isolated from the supernatant of MSCs using a gradient centrifugation method. Administration of MSC-Exo through intramyocardial injection after myocardial ischemia reperfusion reduced infarct size and alleviated inflammation level in heart and serum. Systemic depletion of macrophages with clodronate liposomes abolished the curative effects of MSC-Exo. MSC-Exo modified the polarization of M1 macrophages to M2 macrophages both in vivo and in vitro. miRNA-sequencing of MSC-Exo and bioinformatics analysis implicated miR-182 as a potent candidate mediator of macrophage polarization and TLR4 as a downstream target. Diminishing miR-182 in MSC-Exo partially attenuated its modulation of macrophage polarization. Likewise, knock down of TLR4 also conferred cardioprotective efficacy and reduced inflammation level in a mouse model of myocardial ischemia/reperfusion. Conclusion: Our data indicates that MSC-Exo attenuates myocardial ischemia/reperfusion injury via shuttling miR-182 that modifies the polarization state of macrophages. This study sheds new light on the application of MSC-Exo a potential therapeutic tool for myocardial ischemia/reperfusion injury.
Project description:The human HEK293 / 293T and rat cardiomyoblast H9c2 cell lines are commonly employed for microRNA-mRNA interaction studies. Here, I provide microRNA sequencing data obtained from each of these lines to better document which microRNAs are endogenously expressed at high or low levels. Small RNA sequencing profiles were generated from cultured HEK293 and H9c2 cells on Illumina HiSeq 2000 instruments.
Project description:During culture, H9c2 cells acquire a myotubule phenotype where a critical component is the inclusion of retinoic acid (RA). The results from some authors on H9c2 suggested that thousands of genes respond to RA stimuli, while other authors report hundreds of genes responding to RA over different cell types. We investigated the response to RA in H9c2 cells controlling for culture time.
Project description:Breast milk is a complex liquid that enriched in immunological components and affect the development of the infant immune system. Exosomes, the membranous vesicles of endocytic origin, are ubiquitously in various body fluids which can mediate intercellular communication. MicroRNAs (miRNAs), a well-defined group of non-coding small RNAs, in human breast milk are packaged inside exosomes. Here, we present the identification of miRNAs in human breast milk exosomes using deep sequencing technology. We found that the immune-related miRNAs are enriched in breast milk exosomes, and are resistant to the general harsh conditions. Four small RNA libraries in human breast milk exosomes from four healthy women (30 +/- 0.9 years old, primiparity) when the infant were aged at 60 days were sequenced.
Project description:Analysis of H9C2 cells following PAGln treatment or not. PAGln regulates various mRNA expression in H9C2 cells.Results provide insight into the role of PAGln-involved mechanisms underlying PAGln-mediated effects on coronary artery disease (CAD).