PMID- 28086912 OWN - NLM STAT- MEDLINE DCOM- 20171013 LR - 20181202 IS - 1742-2094 (Electronic) IS - 1742-2094 (Linking) VI - 14 IP - 1 DP - 2017 Jan 10 TI - WWL70 attenuates PGE(2) production derived from 2-arachidonoylglycerol in microglia by ABHD6-independent mechanism. PG - 7 LID - 10.1186/s12974-016-0783-4 [doi] LID - 7 AB - BACKGROUND: alpha/beta-Hydrolase domain 6 (ABHD6) is one of the major enzymes for endocannabinoid 2-arachidonoylglycerol (2-AG) hydrolysis in microglia cells. Our recent studies have shown that a selective ABHD6 inhibitor WWL70 has anti-inflammatory and neuroprotective effects in animal models of traumatic brain injury and multiple sclerosis. However, the role of ABHD6 in the neuroinflammatory response and the mechanisms by which WWL70 suppresses inflammation has not yet been elucidated in reactive microglia. METHODS: The hydrolytic activity and the levels of 2-AG in BV2 cells were measured by radioactivity assay and liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS). The expression of cyclooxygenase-2 (COX-2) and prostaglandin E(2) (PGE(2)) synthases in microglia treated with lipopolysaccharide (LPS) with/without WWL70 was determined by western blot and quantitative RT-PCR. The conversion of 2-AG to PGE(2) or PGE(2)-glyceryl ester (PGE(2)-G) was assessed by enzyme-linked immunoassay (EIA) or LC-MS/MS. The involvement of ABHD6 in PGE(2) production was assessed using pharmacological inhibitors and small interfering RNA (siRNA). The effect of WWL70 on PGE(2) biosynthesis activity in the microsome fraction from BV2 cells and experimental autoimmune encephalopathy (EAE) mouse brain was also examined. RESULTS: We found that WWL70 suppressed PGE(2) production in LPS-activated microglia via cannabinoid receptor-independent mechanisms, although intracellular levels of 2-AG were elevated by WWL70 treatment. This reduction was not attributable to WWL70 inhibition of ABHD6, given the fact that downregulation of ABHD6 by siRNA or use of KT182, an alternative ABHD6 inhibitor failed to suppress PGE(2) production. WWL70 attenuated the expression of COX-2 and PGES-1/2 leading to the downregulation of the biosynthetic pathways of PGE(2) and PGE(2)-G. Moreover, PGE(2) production from arachidonic acid was reduced in the microsome fraction, indicating that WWL70 also targets PGE(2) biosynthetic enzymes, which are likely to contribute to the therapeutic mechanisms of WWL70 in the EAE mouse model. CONCLUSIONS: WWL70 is an anti-inflammatory therapeutic agent capable of inhibiting PGE(2) and PGE(2)-G production, primarily due to its reduction of COX-2 and microsomal PGES-1/2 expression and their PGE(2) biosynthesis activity in microglia cells, as well as in the EAE mouse brain. FAU - Tanaka, Mikiei AU - Tanaka M AD - Department of Anatomy, Physiology and Genetics, Uniformed Services University of the Health Sciences, 4301 Jones Bridge Road, Bethesda, MD, 20814, USA. FAU - Moran, Sean AU - Moran S AD - Biomedical Instrumentation Center, Uniformed Services University of the Health Sciences, 4301 Jones Bridge Road, Bethesda, MD, 20814, USA. FAU - Wen, Jie AU - Wen J AD - Department of Anatomy, Physiology and Genetics, Uniformed Services University of the Health Sciences, 4301 Jones Bridge Road, Bethesda, MD, 20814, USA. FAU - Affram, Kwame AU - Affram K AD - Department of Pharmacology and Molecular Therapeutics, Uniformed Services University of the Health Sciences, 4301 Jones Bridge Road, Bethesda, MD, 20814, USA. AD - Neuroscience Program, Uniformed Services University of the Health Sciences, 4301 Jones Bridge Road, Bethesda, MD, 20814, USA. FAU - Chen, Tinghua AU - Chen T AD - Department of Anatomy, Physiology and Genetics, Uniformed Services University of the Health Sciences, 4301 Jones Bridge Road, Bethesda, MD, 20814, USA. FAU - Symes, Aviva J AU - Symes AJ AD - Department of Pharmacology and Molecular Therapeutics, Uniformed Services University of the Health Sciences, 4301 Jones Bridge Road, Bethesda, MD, 20814, USA. AD - Neuroscience Program, Uniformed Services University of the Health Sciences, 4301 Jones Bridge Road, Bethesda, MD, 20814, USA. FAU - Zhang, Yumin AU - Zhang Y AD - Department of Anatomy, Physiology and Genetics, Uniformed Services University of the Health Sciences, 4301 Jones Bridge Road, Bethesda, MD, 20814, USA. yumin.zhang@usuhs.edu. AD - Neuroscience Program, Uniformed Services University of the Health Sciences, 4301 Jones Bridge Road, Bethesda, MD, 20814, USA. yumin.zhang@usuhs.edu. LA - eng PT - Journal Article DEP - 20170110 PL - England TA - J Neuroinflammation JT - Journal of neuroinflammation JID - 101222974 RN - 0 (Arachidonic Acids) RN - 0 (Biphenyl Compounds) RN - 0 (Cannabinoid Receptor Agonists) RN - 0 (Cannabinoid Receptor Antagonists) RN - 0 (Carbamates) RN - 0 (Endocannabinoids) RN - 0 (Enzyme Inhibitors) RN - 0 (Glycerides) RN - 0 (Lipopolysaccharides) RN - 0 (N-methyl-N-((3-(4-pyridinyl)phenyl)methyl)-4'-(aminocarbonyl)(1,1'-biphenyl)-4-yl ester, carbamic acid) RN - 0 (Piperidines) RN - 0 (Pyrazoles) RN - 0 (RNA, Small Interfering) RN - 8D239QDW64 (glyceryl 2-arachidonate) RN - EC 1.14.99.1 (Cyclooxygenase 2) RN - EC 3.1.1.23 (ABHD6 protein, mouse) RN - EC 3.1.1.23 (Monoacylglycerol Lipases) RN - K7Q1JQR04M (Dinoprostone) RN - RML78EN3XE (Rimonabant) SB - IM MH - Animals MH - Arachidonic Acids/*metabolism MH - Biphenyl Compounds/*pharmacology MH - Cannabinoid Receptor Agonists/pharmacology MH - Cannabinoid Receptor Antagonists/pharmacology MH - Carbamates/*pharmacology MH - Cells, Cultured MH - Cerebral Cortex/cytology MH - Cyclooxygenase 2/metabolism MH - Dinoprostone/genetics/*metabolism MH - Endocannabinoids/*metabolism MH - Enzyme Inhibitors/*pharmacology MH - Female MH - Glycerides/*metabolism MH - Hydrolysis/drug effects MH - Lipopolysaccharides/pharmacology MH - Male MH - Mice MH - Microglia/*drug effects/metabolism MH - Monoacylglycerol Lipases/*metabolism MH - Piperidines/pharmacology MH - Pyrazoles/pharmacology MH - RNA, Small Interfering/genetics/metabolism MH - Rats MH - Rats, Sprague-Dawley MH - Rimonabant PMC - PMC5234251 OTO - NOTNLM OT - 2-AG OT - ABHD6 OT - Arachidonic acid OT - BV2 cells OT - COX-2 OT - Endocannabinoid OT - Microglia OT - PGE synthase OT - PGE2 OT - PGE2-glyceryl ester EDAT- 2017/01/15 06:00 MHDA- 2017/10/14 06:00 PMCR- 2017/01/10 CRDT- 2017/01/15 06:00 PHST- 2016/07/08 00:00 [received] PHST- 2016/12/22 00:00 [accepted] PHST- 2017/01/15 06:00 [entrez] PHST- 2017/01/15 06:00 [pubmed] PHST- 2017/10/14 06:00 [medline] PHST- 2017/01/10 00:00 [pmc-release] AID - 10.1186/s12974-016-0783-4 [pii] AID - 783 [pii] AID - 10.1186/s12974-016-0783-4 [doi] PST - epublish SO - J Neuroinflammation. 2017 Jan 10;14(1):7. doi: 10.1186/s12974-016-0783-4.