The role of morphine on rat neural stem cells viability, neuro-angiogenesis and neuro-steroidgenesis properties

A lack of comprehensive data exists on the effect of morphine on neural stem cell neuro-steroidogenesis and neuro-angiogenesis properties. We, herein, investigated the effects of morphine (100 mu M), naloxone (100 mu M) and their combination on rat neural stem cells viability, clonogenicity and Ki-67 expression over a period of 72 h. Any alterations in the total fatty acids profile under treatment protocols were elucidated by direct transesterification method. We also monitored the expression of p53, aromatase and 5-alpha reductase by real-time PCR assay. To examine angiogenic capacity, in vitro tubulogenesis and the level of VE-cadherin transcript were investigated during neural to endothelial differentiation under the experimental procedure. Cells supplemented with morphine displayed reduced survival (p < 0.01) and clonogenicity (p < 0.001). Flow cytometric analysis showed a decrease in Ki-67 during 72 h. Naloxone potentially blunted morphine induced all effects. The normal levels of fatty acids, including saturated and unsaturated were altered by naloxone and morphine supplements. Following 48 h, the up-regulation of p53, aromatase and 5-alpha reductase genes occurred in morphine-primed cells. Using three-dimensional culture models of angiogenesis and real time PCR assay, we showed morphine impaired the tubulogenesis properties of neural stem cells (p < 0.001) by the inhibition of trans-differentiation into vascular cells and led to decrease of in VE-cadherin expression. Collectively, morphine strongly impaired the healthy status of neural stem cells by inducing p53 and concurrent elevation of aromatase and 5-alpha reductase activities especially during early 48 h. Also, neural stem cells-being exposed to morphine lost their potency to elicit angiogenesis. (C) 2016 Elsevier Ireland Ltd. All rights reserved.