Spinal phospholipase A2 in inflammatory hyperalgesia: role of group IVA cPLA2.
Current work has shown the importance of spinal cyclooxygenase (COX) products in facilitatory processes leading to tissue injury induced hyperalgesia. This cascade must originate with free arachidonic acid (AA) released by the activity of spinal phospholipase A2s (PLA2). In the present work, we studied the role of PLA2s in spinal sensitization. We first demonstrate the presence of constitutive mRNA in the spinal cord for PLA2 Groups IB, IIA, IIC, IVA, V and VI by reverse transcription-polymerase chain reaction (RT-PCR) and sequencing. Using quantitative-PCR, we found that Group IVA cPLA2 and Group VI iPLA2 are the predominant PLA2 messages in the spinal cord. Western blotting and activity assays specific for Group IVA cPLA2 and Group VI iPLA2 verified the presence of these enzymes. PLA2 activity in spinal cord homogenates was suppressed by methyl arachidonyl fluorophosphonate (MAFP) and arachidonyl trifluoromethylketone (AACOCF3), mixed inhibitors of Group IVA cPLA2 and Group VI iPLA2 as well as by bromoenol lactone (BEL), a Group VI iPLA2 inhibitor. The spinal expression of PLA2 mRNA or protein was not altered in the face of peripheral inflammation. Secondly, we showed that intrathecal (i.t.) administration of MAFP and AACOCF3, but not BEL, dose-dependently prevented thermal hyperalgesia induced by intraplantar carrageenan as well as formalin-induced flinching. Finally, i.t. injection of AACOCF3, at antihyperalgesic doses, decreased the release of prostaglandin E2 (PGE2) into spinal dialysate evoked by i.t. NMDA, while i.t. injection of BEL had no effect. Taken together, this work points to a role for constitutive Group IVA cPLA2 in spinal nociceptive processing.
Lucas
Karin Killermann
KK
Department of Chemistry, School of Medicine, University of California, 9500 Gilman Drive, San Diego, La Jolla, CA 92093-0601, USA.
Svensson
Camilla I
CI
Hua
Xiao-Ying
XY
Yaksh
Tony L
TL
Dennis
Edward A
EA
D000818:Animals; D004305:Dose-Response Relationship, Drug; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects / Q000502:physiology; D054513:Group IV Phospholipases A2; D006930:Hyperalgesia / Q000201:enzymology* / Q000235:genetics; D008297:Male; D016202:N-Methylaspartate / Q000494:pharmacology; D010147:Pain Measurement / Q000187:drug effects / Q000379:methods; D010741:Phospholipases A / Q000096:biosynthesis* / Q000235:genetics; D054467:Phospholipases A2; D051381:Rats; D017207:Rats, Sprague-Dawley; D013116:Spinal Cord / Q000201:enzymology*
D016202:N-Methylaspartate; D010741:Phospholipases A; D054513:Group IV Phospholipases A2; D054467:Phospholipases A2; C516391:Pla2g4a protein, rat