title: Activation of cannabinoid receptor 2 attenuates leukocyte-endothelial cell interactions and blood-brain barrier dysfunction under inflammatory conditions.
abstract: Previous studies have shown that modulation of the receptor-mediated cannabinoid system during neuroinflammation can produce potent neuroprotective and anti-inflammatory effects. However, in this context, little is known about how selective activation of the cannabinoid type-2 receptor (CB2R) affects the activated state of the brain endothelium and blood-brain barrier (BBB) function. Using human brain tissues and primary human brain microvascular endothelial cells (BMVECs), we demonstrate that the CB2R is highly upregulated during inflammatory insult. We then examined whether the CB2R agonists could attenuate inflammatory responses at the BBB using a mouse model of LPS-induced encephalitis and highly selective CB2R agonists. Visualization by intravital microscopy revealed that administration of JWH133 [(6aR,10aR)-3-(1,1-dimethylbutyl)-6a,7,10,10a-tetrahydro-6,6,9-trimethyl-6H-dibenzo[b,d]pyran] or a novel resorcinol-based compound, O-1966 (1-[4-(1,1-dimethyl-heptyl)-2,6-dimethoxy-phenyl]-3-methyl-cyclohexanol), greatly attenuated leukocyte adhesion in surface pial vessels and in deep ascending cortical postcapillary venules. BBB permeability assessments with small and large fluorescent tracers showed that CB2R agonists were effective at preventing barrier leakiness after LPS administration. To determine whether the effects by CB2R agonists on barrier protection are not only due to the CB2R modulation of immune cell function, we tested the agonists in vitro with barrier-forming primary BMVECs. Remarkably, the addition of CB2R agonist increased transendothelial electrical resistance and increased the amount of tight junction protein present in membrane fractions. Furthermore, CB2R agonists decreased the induction of intercellular adhesion molecule-1 and vascular cell adhesion molecule-1 surface expression in BMVECs exposed to various proinflammatory mediators. Together, these results suggest that pharmacological CB2R ligands offer a new strategy for BBB protection during neuroinflammation.
authors.affiliation: Department of Pathology and Laboratory Medicine, Temple University School of Medicine, Philadelphia, PA 19140, USA.
authors.forename: Servio H
authors.initials: SH
authors.lastname: Ramirez
authors.forename: János
authors.initials: J
authors.lastname: Haskó
authors.forename: Andrew
authors.initials: A
authors.lastname: Skuba
authors.forename: Shongshan
authors.initials: S
authors.lastname: Fan
authors.forename: Holly
authors.initials: H
authors.lastname: Dykstra
authors.forename: Ryan
authors.initials: R
authors.lastname: McCormick
authors.forename: Nancy
authors.initials: N
authors.lastname: Reichenbach
authors.forename: Istvan
authors.initials: I
authors.lastname: Krizbai
authors.forename: Anu
authors.initials: A
authors.lastname: Mahadevan
authors.forename: Ming
authors.initials: M
authors.lastname: Zhang
authors.forename: Ronald
authors.initials: R
authors.lastname: Tuma
authors.forename: Young-Jin
authors.initials: YJ
authors.lastname: Son
authors.forename: Yuri
authors.initials: Y
authors.lastname: Persidsky
mesh_terms: D000818:Animals; D000840:Anisoles / Q000494:pharmacology / Q000627:therapeutic use; D001812:Blood-Brain Barrier / Q000187:drug effects / Q000503:physiopathology*; D001892:Camphanes / Q000494:pharmacology; D002186:Cannabinoids / Q000494:pharmacology; D002199:Capillary Permeability / Q000187:drug effects / Q000502:physiology; D002448:Cell Adhesion / Q000187:drug effects / Q000235:genetics; D002478:Cells, Cultured; D003511:Cyclohexanols; D003911:Dextrans / Q000378:metabolism; D004195:Disease Models, Animal; D017097:Electric Impedance; D004660:Encephalitis / Q000139:chemically induced / Q000473:pathology*; D042783:Endothelial Cells / Q000502:physiology*; D004727:Endothelium / Q000378:metabolism; D005434:Flow Cytometry; D016650:Fluorescein-5-isothiocyanate / Q000031:analogs & derivatives / Q000378:metabolism; D005786:Gene Expression Regulation / Q000187:drug effects / Q000235:genetics; D006801:Humans; D018799:Intercellular Adhesion Molecule-1 / Q000378:metabolism; D007962:Leukocytes / Q000502:physiology*; D008070:Lipopolysaccharides / Q000009:adverse effects; D008565:Membrane Proteins / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D010750:Phosphoproteins / Q000378:metabolism; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000378:metabolism; D011720:Pyrazoles / Q000494:pharmacology; D043885:Receptor, Cannabinoid, CB2 / Q000819:agonists / Q000037:antagonists & inhibitors / Q000172:deficiency / Q000378:metabolism*; D018709:Statistics, Nonparametric; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology; D019010:Vascular Cell Adhesion Molecule-1 / Q000378:metabolism; D062826:Zonula Occludens-1 Protein
chemical_list: C531961:1-(4-(1,1-dimethylheptyl)-2,6-dimethoxyphenyl)-3-methylcyclohexanol; D000840:Anisoles; D001892:Camphanes; D002186:Cannabinoids; D003511:Cyclohexanols; D003911:Dextrans; D008070:Lipopolysaccharides; D008565:Membrane Proteins; D010750:Phosphoproteins; D019408:Platelet Endothelial Cell Adhesion Molecule-1; D011720:Pyrazoles; D043885:Receptor, Cannabinoid, CB2; C110630:SR 144528; C569042:TJP1 protein, human; C569043:Tjp1 protein, mouse; D014409:Tumor Necrosis Factor-alpha; D019010:Vascular Cell Adhesion Molecule-1; D062826:Zonula Occludens-1 Protein; C015219:fluorescein isothiocyanate dextran; D018799:Intercellular Adhesion Molecule-1; D016650:Fluorescein-5-isothiocyanate; C432747:1,1-dimethylbutyl-1-deoxy-Delta(9)-THC