title: The non-psychoactive plant cannabinoid, cannabidiol affects cholesterol metabolism-related genes in microglial cells.
abstract: Cannabidiol (CBD) is a non-psychoactive plant cannabinoid that is clinically used in a 1:1 mixture with the psychoactive cannabinoid Δ(9)-tetrahydrocannabinol (THC) for the treatment of neuropathic pain and spasticity in multiple sclerosis. Our group previously reported that CBD exerts anti-inflammatory effects on microglial cells. In addition, we found that CBD treatment increases the accumulation of the endocannabinoid N-arachidonoyl ethanolamine (AEA), thus enhancing endocannabinoid signaling. Here we proceeded to investigate the effects of CBD on the modulation of lipid-related genes in microglial cells. Cell viability was tested using FACS analysis, AEA levels were measured using LC/MS/MS, gene array analysis was validated with real-time qPCR, and cytokine release was measured using ELISA. We report that CBD significantly upregulated the mRNAs of the enzymes sterol-O-acyl transferase (Soat2), which synthesizes cholesteryl esters, and of sterol 27-hydroxylase (Cyp27a1). In addition, CBD increased the mRNA of the lipid droplet-associated protein, perilipin2 (Plin2). Moreover, we found that pretreatment of the cells with the cholesterol chelating agent, methyl-β-cyclodextrin (MBCD), reversed the CBD-induced increase in Soat2 mRNA but not in Plin2 mRNA. Incubation with AEA increased the level of Plin2, but not of Soat2 mRNA. Furthermore, MBCD treatment did not affect the reduction by CBD of the LPS-induced release of the proinflammatory cytokine IL-1β. CBD treatment modulates cholesterol homeostasis in microglial cells, and pretreatment with MBCD reverses this effect without interfering with CBD's anti-inflammatory effects. The effects of the CBD-induced increase in AEA accumulation on lipid-gene expression are discussed.
authors.affiliation: The Department of Neurobiology, Weizmann Institute of Science, Rehovot, Israel. neta.rimmerman@gmail.com
authors.forename: Neta
authors.initials: N
authors.lastname: Rimmerman
authors.forename: Ana
authors.initials: A
authors.lastname: Juknat
authors.forename: Ewa
authors.initials: E
authors.lastname: Kozela
authors.forename: Rivka
authors.initials: R
authors.lastname: Levy
authors.forename: Heather B
authors.initials: HB
authors.lastname: Bradshaw
authors.forename: Zvi
authors.initials: Z
authors.lastname: Vogel
mesh_terms: D000818:Animals; D001095:Arachidonic Acids / Q000378:metabolism; D002185:Cannabidiol / Q000494:pharmacology*; D016923:Cell Death / Q000187:drug effects; D002460:Cell Line; D002784:Cholesterol / Q000378:metabolism*; D016895:Culture Media, Serum-Free; D013759:Dronabinol / Q000494:pharmacology; D063388:Endocannabinoids; D005786:Gene Expression Regulation / Q000187:drug effects; D053583:Interleukin-1beta / Q000378:metabolism; D050356:Lipid Metabolism / Q000187:drug effects* / Q000235:genetics*; D008070:Lipopolysaccharides / Q000494:pharmacology; D051379:Mice; D017628:Microglia / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism*; D020411:Oligonucleotide Array Sequence Analysis; D010944:Plants / Q000737:chemistry*; D053284:Polyunsaturated Alkamides; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D013997:Time Factors; D047392:beta-Cyclodextrins / Q000494:pharmacology
chemical_list: D001095:Arachidonic Acids; D016895:Culture Media, Serum-Free; D063388:Endocannabinoids; D053583:Interleukin-1beta; D008070:Lipopolysaccharides; D053284:Polyunsaturated Alkamides; D012333:RNA, Messenger; D047392:beta-Cyclodextrins; C108732:methyl-beta-cyclodextrin; D002185:Cannabidiol; D013759:Dronabinol; D002784:Cholesterol; C078814:anandamide