Homo sapiens
Ontology highlight
ABSTRACT:
PROVIDER: PRJNA75939 | ENA |
REPOSITORIES: ENA
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Project description:In recent years, a critical clinical problem that cannot be ignored is the incidence of venous thrombosis gradually increasing. Although many animal models of thrombosis have been established, the complex mechanism of thrombosis has not been elucidated. We successfully established a new animal model of venous thrombosis by infrared-pulse laser selectively targeting damage to the venous endothelium of zebrafish tail, resulting in the aggregation of red blood cells and platelets at the site of injury, forming a thrombus, like the formation of thrombi caused by damaged human venous endothelium. o-Dianisidine staining showed increased hemoglobin density at the injury site and decreased hemoglobin density at the heart site. Utilizing laser microdissection technology, we targeted the acquisition of localized thrombus cell clusters for high-throughput transcriptome sequencing. The data were further analyzed through gene set variation analysis (GSVA) and gene set enrichment analysis (GSEA), with the transcriptome data being examined against backgrounds of GO, KEGG, Reactome, and WP databases. Combining these analyses with molecular biology techniques such as RT-qPCR, WISH, and Western Blot, we observed that, compared to the normal group, macrophages were activated, and erythrocyte differentiation was more vigorous post-thrombus formation. Signaling pathways related to cell adhesion and leukocyte migration were activated, and the expression of inflammatory cytokines increased. Notably, IL-6 and TNF-α significantly increased at the thrombus site, while other cytokines such as IL-1β, IL-10, P-selectin, STAT3, phosphorylated STAT3, p65, and phosphorylated p65 were major players in the inflammatory response during venous thrombosis. The venous thrombosis model established in this study allows a high degree of visualization of thrombosis and provides a feasible and powerful protocol for studying the mechanism of thrombosis. This study may serve as a new venous thrombosis model for exploring the detailed kinetics and underlying mechanisms of thrombosis formation.
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Project description:RNA extracted at 240min. Samples are rRNA depleted. Expression measurement of Homo sapiens genes.
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Project description:Leukocyte flux contributes to thrombus formation in deep veins under pathologic conditions, but mechanisms which inhibit venous thrombosis are incompletely understood. Ectonucleotide di(tri)phosphohydrolase 1 (ENTPD1 or Cd39), an ectoenzyme which catabolizes extracellular adenine nucleotides, is embedded on the surface of endothelial cells and leukocytes. We hypothesized that under venous stasis conditions CD39 regulates inflammation at the vein:blood interface in a murine model of deep vein thrombosis. Gene expression profiling of WT and Cd39-null mice revealed 76 differentially-expressed inflammatory genes that were significantly upregulated in Cd39-deleted mice after venous thrombosis; and validation experiments confirmed high expression of several key inflammatory mediators.
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