title: Morphine treatment enhances extracellular ATP enzymolysis and adenosine generation in rat astrocytes.
abstract: Recent studies have shown that astrocytes play important roles in ATP degradation and adenosine (a well known analgesic molecule) generation, which are closely related to pain signaling pathway. The aim of this study was to investigate whether morphine, a well known analgesic drug, could affect the speeds of ATP enzymolysis and adenosine generation in rat astrocytes. Intracellular calcium concentration ([Ca(2+)](i)) of astrocyte was measured by flow cytometry, and the time points that morphine exerted notable effects were determined for subsequent experiments. Cultured astrocytes were pre-incubated with morphine (1 μmol/L) and then were incubated with substrates, ATP and AMP, for 30 min. The speeds of ATP enzymolysis and adenosine generation were measured by high performance liquid chromatography (HPLC). The results showed that both 1.5 and 48 h of morphine pre-incubation induced maximal ATP enzymolysis speed in astrocytes among all the time points, and there was no statistical difference of ATP enzymolysis speed between morphine treatments for 1.5 and 48 h. As to adenosine, morphine pre-incubation for 1.5 h statistically increased adenosine generation, which was degraded from AMP, in cultured astrocytes compared with control group. However, no difference of adenosine generation was observed after 48 h of morphine pre-incubation. These results indicate that treatment of morphine in vitro dynamically changes the concentrations of ATP and adenosine in extracellular milieu of astrocytic cells. In addition, astrocyte can be regarded as at least one of the target cells of morphine to induce changes of ATP and adenosine levels in central nervous system.
authors.affiliation: Department of Physiology, College of Fundamental Medical Science, Guangzhou University of Chinese Medicine, Guangzhou 510006, China. weiliu1980@yahoo.com
authors.forename: Wei
authors.initials: W
authors.lastname: Liu
authors.forename: Zhan-li
authors.initials: ZL
authors.lastname: Yang
authors.forename: Le-quan
authors.initials: LQ
authors.lastname: Zhou
authors.forename: Xiao-ying
authors.initials: XY
authors.lastname: Li
authors.forename: Fu-man
authors.initials: FM
authors.lastname: Yan
authors.forename: Li
authors.initials: L
authors.lastname: Guan
authors.forename: Hai-mei
authors.initials: HM
authors.lastname: Liu
authors.forename: Jian-qiang
authors.initials: JQ
authors.lastname: Feng
mesh_terms: D000241:Adenosine / Q000096:biosynthesis*; D000255:Adenosine Triphosphate / Q000378:metabolism*; D000701:Analgesics, Opioid / Q000494:pharmacology; D000818:Animals; D000831:Animals, Newborn; D001253:Astrocytes / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism*; D002118:Calcium / Q000032:analysis / Q000378:metabolism; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000166:cytology; D009020:Morphine / Q000494:pharmacology*; D051381:Rats; D017207:Rats, Sprague-Dawley
chemical_list: D000701:Analgesics, Opioid; D009020:Morphine; D000255:Adenosine Triphosphate; D000241:Adenosine; D002118:Calcium