title: The role of morphine on rat neural stem cells viability, neuro-angiogenesis and neuro-steroidgenesis properties.
abstract: 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 (100μM), naloxone (100μM) and their combination on rat neural stem cells viability, clonogenicity and Ki-67 expression over a period of 72h. 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 72h. 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 48h, 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 48h. Also, neural stem cells-being exposed to morphine lost their potency to elicit angiogenesis.
authors.affiliation: Department of Biochemistry and Clinical Laboratories, Faculty of Medicine, Tabriz University of Medical sciences, Tabriz, Iran; Stem Cell Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.
authors.forename: Nima
authors.initials: N
authors.lastname: Abdyazdani
authors.affiliation: Department of Biochemistry and Clinical Laboratories, Faculty of Medicine, Tabriz University of Medical sciences, Tabriz, Iran; Stem Cell Research Center, Tabriz University of Medical Sciences, Tabriz, Iran. Electronic address: alinour65@gmail.com.
authors.forename: Alireza
authors.initials: A
authors.lastname: Nourazarian
authors.affiliation: Stem Cell Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.
authors.forename: Hojjatollah
authors.initials: H
authors.lastname: Nozad Charoudeh
authors.affiliation: Stem Cell Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.
authors.forename: Masoumeh
authors.initials: M
authors.lastname: Kazemi
authors.affiliation: Department of Biochemistry and Clinical Laboratories, Faculty of Medicine, Tabriz University of Medical sciences, Tabriz, Iran; Stem Cell Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.
authors.forename: Navid
authors.initials: N
authors.lastname: Feizy
authors.affiliation: Department of Biochemistry and Clinical Laboratories, Faculty of Medicine, Tabriz University of Medical sciences, Tabriz, Iran; Stem Cell Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.
authors.forename: Maryam
authors.initials: M
authors.lastname: Akbarzade
authors.affiliation: Liver and Gastrointestinal Diseases Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.
authors.forename: Amir
authors.initials: A
authors.lastname: Mehdizadeh
authors.affiliation: Department of Biology, Faculty of Science, Urmia University, Urmia, Iran.
authors.forename: Jafar
authors.initials: J
authors.lastname: Rezaie
authors.affiliation: Stem Cell Research Center, Tabriz University of Medical Sciences, Tabriz, Iran; Department of Applied Cell Sciences, Faculty of Advanced Medical Sciences, Tabriz University of Medical Sciences, Tabriz, Iran. Electronic address: Rezarahbardvm@gmail.com.
authors.forename: Reza
authors.initials: R
authors.lastname: Rahbarghazi
mesh_terms: D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects*; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D042783:Endothelial Cells / Q000187:drug effects / Q000378:metabolism; D009020:Morphine / Q000494:pharmacology*; D009270:Naloxone / Q000494:pharmacology*; D009389:Neovascularization, Pathologic / Q000473:pathology; D058953:Neural Stem Cells / Q000166:cytology* / Q000187:drug effects* / Q000378:metabolism; D051381:Rats
chemical_list: D009270:Naloxone; D009020:Morphine
keywords: Aromatase and 5-alpha reductase expression; Endothelial differentiation; Morphine; Rat neural stem cells; VE-cadherin