| Resumo: | <p>Hypoxia induced factor (HIF) transcriptional complex is a potent inducer of cytokines that enhances therapeutic potential of stem cell. Here, ethyl 2-(2,3-dihyroxybenzamido) acetate (EDBA) was used to stabilize HIF-α in cardiosphere-derived cells (CDCs) with the aim of enhancing their therapeutic potential for myocardial infarction. In this study, neonatal Sprague Dawley (SD) rat hearts (n=12) were excised, minced and cultured to generate explant-derived cells (EDCs) which were subsequently expanded into monolayer cardiosphere-derived cells (CDCs). CDCs were treated with 0.5mM EDBA for 24h before RNA (n=4), protein (n=4) and culture medium (n=4) were collected for genomic, proteomic and metabolic analysis. All data were presented as mean ± standard deviation, SD. EDBA-treated CDCs showed significant increased HIF-1α mRNA expression (2.4 ± 0.6-fold, p < 0.01) after 24 hours, compared with non-treated control cells. EDBA significantly increased cardiac stem cell marker, c-Kit (1.5 ± 0.2-fold, p < 0.05) and reduced the cardiac mesenchymal cell markers, CD90 and CD105 (both 0.7 ± 0.2-fold, p < 0.05), generating a culture containing higher c-Kit+ cardiac stem cell population. EDBA treatment also significantly increased the expression of cytokines (p < 0.05) involved in stem cell trafficking (CXCR4, 5.1 ± 2.0-fold), erythropoiesis (EPO, 2.2 ± 0.7-fold) and angiogenesis (VEGF, 2.0 ± 0.6-fold), enhancing the therapeutic potential of CDCs for myocardial infarction. Of note, EDBA significantly increased glucose uptake (2.4 ± 0.2-fold, p < 0.05) and lactate accumulation (2.0 ± 0.1-fold, p < 0.01) in the CDC culture medium, suggesting increased glycolytic metabolism. Further, EDBA significantly reduced oxygen consumption in CDC by 80% (p < 0.05), suggesting that treated cells could survive better after transplantation into the hypoxic infarcted myocardium. In conclusion, EDBA stabilized HIF-1α, which induced metabolic changes in the CDCs. This work could impact on cardiac stem cell therapy for myocardial infarction.</p>
|
|---|