OBJECTIVE Endocrine disruptor chemicals (EDCs) such as Tributyltin (TBT) are toxic compounds widely present in the environment with harmful effects including perturbation of transmembrane ion gradients, oxidative stress and apoptosis. TBT may constitute a risk factor for cardiovascular disease inducing morphological vascular alteration and impairing coronary vascular reactivity. Recently we described the isolation of porcine Aortic Vascular Precursors Cells (pAVPCs), these cells have the phenotypic characteristics of mesenchymal cells, and functional properties of pericytes. Few studies have investigated the TBT effect on stem cells and none has been done on vascular precursor cells. Aim of the present study was to evaluate the influence of TBT on pAVPCs gene expression. MATERIALS AND METHODS pAVPCs were seeded in a 24 wells (3x105/well) plate and exposed to increasing doses of TBT (10, 100, 500 nM) for 48h, cytotoxicity was evaluated and gene expression of mesenchymal (CD90, CD73, CD105), perycite (NG2, Nestin, PDGFβR, alpha-SMA) angiogenetic (VEGF, Flt-1) and stem cell (NANOG) markers were investigated by qPCR analysis. RESULTS: TBT reduced cell viability reaching a mortality rate of approximately 30% at the highest dose, furthermore TBT influenced the expression of most of the studied genes. Mesenchymal markers increased at the lowest dose (10 nM) and decrease at the highest dose (500nM), otherwise the expression of NG2, Nestin and PDGFβR decreased at all three doses studied. The most sensitive gene was alpha-SMA, with an increase of approximately 7 times at the lowest dose and a strong reduction at the higher dose. Angiogenetic and stem cell markers did not seem affected. CONCLUSIONS Overall TBT perturbed the expression of pAVPCs’ genes pattern, further investigation will be necessary to clarify the structural and functional effects of these alterations and the impact of these changes in relation to differentiation potential.
Alteration of gene expression induced by Tributyltin in porcine Aortic Vascular Precursor Cells
BERNARDINI, CHIARA;ZANNONI, AUGUSTA;BERTOCCHI, MARTINA;FORNI, MONICA
2014
Abstract
OBJECTIVE Endocrine disruptor chemicals (EDCs) such as Tributyltin (TBT) are toxic compounds widely present in the environment with harmful effects including perturbation of transmembrane ion gradients, oxidative stress and apoptosis. TBT may constitute a risk factor for cardiovascular disease inducing morphological vascular alteration and impairing coronary vascular reactivity. Recently we described the isolation of porcine Aortic Vascular Precursors Cells (pAVPCs), these cells have the phenotypic characteristics of mesenchymal cells, and functional properties of pericytes. Few studies have investigated the TBT effect on stem cells and none has been done on vascular precursor cells. Aim of the present study was to evaluate the influence of TBT on pAVPCs gene expression. MATERIALS AND METHODS pAVPCs were seeded in a 24 wells (3x105/well) plate and exposed to increasing doses of TBT (10, 100, 500 nM) for 48h, cytotoxicity was evaluated and gene expression of mesenchymal (CD90, CD73, CD105), perycite (NG2, Nestin, PDGFβR, alpha-SMA) angiogenetic (VEGF, Flt-1) and stem cell (NANOG) markers were investigated by qPCR analysis. RESULTS: TBT reduced cell viability reaching a mortality rate of approximately 30% at the highest dose, furthermore TBT influenced the expression of most of the studied genes. Mesenchymal markers increased at the lowest dose (10 nM) and decrease at the highest dose (500nM), otherwise the expression of NG2, Nestin and PDGFβR decreased at all three doses studied. The most sensitive gene was alpha-SMA, with an increase of approximately 7 times at the lowest dose and a strong reduction at the higher dose. Angiogenetic and stem cell markers did not seem affected. CONCLUSIONS Overall TBT perturbed the expression of pAVPCs’ genes pattern, further investigation will be necessary to clarify the structural and functional effects of these alterations and the impact of these changes in relation to differentiation potential.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
