hu-308 and Inflammation

First isolated from Cannabis in 1940 by Roger Adams, the structure of CBD was not completely elucidated until 1963. Subsequent studies resulted in the pronouncement that THC was the 'active' principle of Cannabis and research then focused primarily on it to the virtual exclusion of CBD. This was no doubt due to the belief that activity meant psychoactivity that was shown by THC and not by CBD. In retrospect this must be seen as unfortunate since a number of actions of CBD with potential therapeutic benefit were downplayed for many years. In this review, attention will be focused on the effects of CBD in the broad area of inflammation where such benefits seem likely to be developed. Topics covered in this review are; the medicinal chemistry of CBD, CBD receptor binding involved in controlling Inflammation, signaling events generated by CBD, downstream events affected by CBD (gene expression and transcription), functional effects reported for CBD and combined THC plus CBD treatment.

Topics: Animals; Cannabidiol; Humans; Inflammation; Inflammation Mediators; Protein Structure, Tertiary; Treatment Outcome

Topics: Acute Lung Injury; Animals; Cannabinoids; Cytokines; Dexamethasone; Endotoxins; Inflammation; Lipopolysaccharides; Lung; Mice; Mice, Inbred C57BL; Microcirculation; Pneumonia; Respiratory Distress Syndrome; Sepsis

Cannabinoid receptor 2 (CB2), whose activities are upregulated during sepsis, may be related to the regulation of inflammatory programmed cell death called pyroptosis. The aim of this study is to investigate the role of CB2 activation in attenuation of inflammation through inhibiting pyroptosis in cecal ligation puncture (CLP)-induced sepsis andlipopolysaccharide (LPS) + ATP-stimulated macrophages.. C57BL/6 mice were subjected to CLP procedure and treated with CB2 agonist HU308 and CB2 antagonist AM630. Lung tissues were collected for analyses of lung W/D ratio, inflammatory factors levels, and pyroptosis-related protein expression. Murine bone-marrow-derived macrophages (BMDM) were treated with LPS and ATP to construct a septic model in vitro in the presence of HU308 and AM630 for assessment of cell injury, cytokine levels and pyroptosis-related protein expression accordingly. To verify the relationship between CB2 receptors and pyroptosis in the process of inflammatory response, BMDM were transduced with CB2 receptors knockdown lentiviral vectors in the presence of HU308 and AM630 for assessment of pyroptosis-related protein expression.. CB2 activation ameliorated the release of inflammatory mediators. The results showed that CLP-induced pyroptosis was elevated, and CB2 agonist HU308 treatment inhibited the pyroptosis activity through a decrease of the protein levels of NLRP3 as well as caspase-1 and GSDMD activation. Similar results were obtained in BMDM after LPS and ATP treatment. Treatment with CB2 knockdown lentiviral particles prevented the HU308-induced decreases in cell pyroptosis, demonstrating that endogenous CB2 receptors are required for the cannabinoid-induced cell protection.. CB2 receptors activation plays a protective role in sepsis through inhibition of pyroptosis. The effect of CB2 receptors against pyroptosis depends on the existence of endogenous CB2 receptors.

Topics: Adenosine Triphosphate; Animals; Cannabinoids; Cecum; Disease Models, Animal; Indoles; Inflammation; Ligation; Lipopolysaccharides; Lung; Macrophages; Male; Mice, Inbred C57BL; Primary Cell Culture; Punctures; Pyroptosis; Receptor, Cannabinoid, CB2; Sepsis

Uveitis is inflammation of the uvea which consists of the iris, ciliary body and the choroid of the eye. Uveitis can lead to impaired vision and is responsible for 10% of all cases of blindness globally. Using an endotoxin-induced uveitis (EIU) rodent model, our previous data implicated the endogenous cannabinoid system (ECS) in the amelioration of many of the components of the inflammatory response. Here, we test the hypothesis that the reduction in inflammatory mediators in the EIU model by the CB

Topics: Animals; Cannabinoids; Endocannabinoids; Endotoxins; Eye; Humans; Inflammation; Leukocytes; Lipid Metabolism; Mice; Mice, Knockout; Neutrophil Infiltration; Neutrophils; Prostaglandins; Receptor, Cannabinoid, CB2; RNA, Messenger; Signal Transduction; Uvea; Uveitis

Activation of cannabinoid receptor 2 (CB2R) has been reported to ameliorate the pathogenesis of experimental autoimmune encephalomyelitis (EAE). In this study, we examined whether autophagy is involved in the beneficial effect of CB2R on EAE and explored the mechanism with a focus on inflammasome activation.. EAE severity was analyzed with clinical score and histological score stained by hematoxylin and eosin or luxol fast blue in spinal cord. Immunoblot analysis was conducted to detect proteins of NOD-like receptor family, pyrin domain containing 3 (NLRP3) inflammasome-related caspase-1 (Casp-1) and the maturation of interleukin (IL)-1β as well as autophagy-related light chain 3 (LC3), and Beciln 1 both in vivo and in vitro. Reverse transcription and real-time PCR were used to detect mRNA of NLRP3, IL-1β and Casp-1. Autophagy-related gene 5 (ATG5)-specific siRNA was transiently transfected in BV2 microglia, and immunofluorescence staining was carried out to detect the expression of NLRP3, caspase recruitment domain (ASC), and pro-caspase-1.. The current data indicated that deleting CB2R decreased the expression of LC3-II/LC3-I ratio, Beclin 1 and increased caspase-1 activation and IL-1β production in the spinal cord of EAE mice, whereas activation of CB2R with a specific agonist HU-308 induced inverse effects. Further study indicated that HU-308 could promote autophagy and inhibit expression and activation of NLRP3 inflammasome in BV2 microglia. Blocking autophagy by ATG5-specific siRNA dismissed the effort of CB2R in mediating NLRP3 inflammasome in vitro.. Collectively, our results demonstrated for the first time that CB2R plays a protective role in EAE through promoting autophagy and inhibiting NLRP3 inflammasome activation.

Topics: Animals; Autophagy; Autophagy-Related Protein 5; Cannabinoid Receptor Agonists; Cannabinoids; Carrier Proteins; Caspase 1; Cells, Cultured; Disease Models, Animal; Encephalomyelitis, Autoimmune, Experimental; Gene Expression Regulation; Inflammation; Interleukin-1beta; Lipopolysaccharides; Mice; Mice, Inbred C57BL; Mice, Knockout; Microglia; Microtubule-Associated Proteins; NLR Family, Pyrin Domain-Containing 3 Protein; Receptor, Cannabinoid, CB2; Spinal Cord

Cisplatin is an important chemotherapeutic agent; however, its nephrotoxicity limits its clinical use. Enhanced inflammatory response and oxidative/nitrosative stress seem to play a key role in the development of cisplatin-induced nephropathy. Activation of cannabinoid-2 (CB(2)) receptors with selective agonists exerts anti-inflammatory and tissue-protective effects in various disease models. We have investigated the role of CB(2) receptors in cisplatin-induced nephrotoxicity using the selective CB(2) receptor agonist HU-308 and CB(2) knockout mice. Cisplatin significantly increased inflammation (leukocyte infiltration, CXCL1/2, MCP-1, TNFalpha, and IL-1beta levels) and expression of adhesion molecule ICAM-1 and superoxide-generating enzymes NOX2, NOX4, and NOX1 and enhanced ROS generation, iNOS expression, nitrotyrosine formation, and apoptotic and poly(ADP-ribose) polymerase-dependent cell death in the kidneys of mice, associated with marked histopathological damage and impaired renal function (elevated serum BUN and creatinine levels) 3 days after the administration of the drug. CB(2) agonist attenuated the cisplatin-induced inflammatory response, oxidative/nitrosative stress, and cell death in the kidney and improved renal function, whereas CB(2) knockouts developed enhanced inflammation and tissue injury. Thus, the endocannabinoid system, through CB(2) receptors, protects against cisplatin-induced kidney damage by attenuating inflammation and oxidative/nitrosative stress, and selective CB(2) agonists may represent a promising novel approach to preventing this devastating complication of chemotherapy.

Topics: Animals; Antineoplastic Agents; Apoptosis; Cannabinoids; Caspases; Cisplatin; In Situ Nick-End Labeling; Inflammation; Kidney Diseases; Kidney Function Tests; Lipid Peroxidation; Male; Mice; Mice, Inbred C57BL; Mice, Knockout; Oxidation-Reduction; Oxidative Stress; Peroxidase; Poly(ADP-ribose) Polymerases; Reactive Oxygen Species; Receptor, Cannabinoid, CB2; Tyrosine

In this study, we have investigated the role of the cannabinoid CB(2) (CB(2)) receptor in an in vivo mouse model of hepatic ischemia/reperfusion (I/R) injury. In addition, we have assessed the role of the CB(2) receptor in TNF-alpha-induced ICAM-1 and VCAM-1 expression in human liver sinusoidal endothelial cells (HLSECs) and in the adhesion of human neutrophils to HLSECs in vitro. The potent CB(2) receptor agonist HU-308, given prior to the induction of I/R, significantly attenuated the extent of liver damage (measured by serum alanine aminotransferase and lactate dehydrogenase) and decreased serum and tissue TNF-alpha, MIP-1alpha, and MIP-2 levels, tissue lipid peroxidation, neutrophil infiltration, DNA fragmentation, and caspase 3 activity. The protective effect of HU-308 against liver damage was also preserved when given right after the ischemic episode. HU-308 also attenuated the TNF-alpha-induced ICAM-1 and VCAM-1 expression in HLSECs, which expressed CB(2) receptors, and the adhesion of human neutrophils to HLSECs in vitro. These findings suggest that selective CB(2) receptor agonists may represent a novel, protective strategy against I/R injury by attenuating oxidative stress, inflammatory response, and apoptosis.

Topics: Alanine Transaminase; Animals; Apoptosis; Cannabinoid Receptor Agonists; Cannabinoids; Cell Adhesion; Chemokines; Endothelial Cells; Humans; Inflammation; Inflammation Mediators; Intercellular Adhesion Molecule-1; Ischemia; L-Lactate Dehydrogenase; Leukocytes; Lipid Peroxidation; Liver; Mice; Oxidative Stress; Peroxidase; Reperfusion Injury; Tumor Necrosis Factor-alpha; Vascular Cell Adhesion Molecule-1

Targeting cannabinoid-2 (CB(2)) receptors with selective agonists may represent a novel therapeutic avenue in various inflammatory diseases, but the mechanisms by which CB(2) activation exerts its anti-inflammatory effects and the cellular targets are elusive. Here, we investigated the effects of CB(2)-receptor activation on TNF-alpha-induced signal transduction in human coronary artery endothelial cells in vitro and on endotoxin-induced vascular inflammatory response in vivo. TNF-alpha induced NF-kappaB and RhoA activation and upregulation of adhesion molecules ICAM-1 and VCAM-1, increased expression of monocyte chemoattractant protein, enhanced transendothelial migration of monocytes, and augmented monocyte-endothelial adhesion. Remarkably, all of the above-mentioned effects of TNF-alpha were attenuated by CB(2) agonists. CB(2) agonists also decreased the TNF-alpha- and/or endotoxin-induced ICAM-1 and VCAM-1 expression in isolated aortas and the adhesion of monocytes to aortic vascular endothelium. CB(1) and CB(2) receptors were detectable in human coronary artery endothelial cells by Western blotting, RT-PCR, real-time PCR, and immunofluorescence staining. Because the above-mentioned TNF-alpha-induced phenotypic changes are critical in the initiation and progression of atherosclerosis and restenosis, our findings suggest that targeting CB(2) receptors on endothelial cells may offer a novel approach in the treatment of these pathologies.

Topics: Animals; Anti-Inflammatory Agents; Aorta; Cannabinoids; Cells, Cultured; Chemokine CCL2; Coronary Vessels; Disease Models, Animal; Dose-Response Relationship, Drug; Endothelial Cells; Humans; Inflammation; Intercellular Adhesion Molecule-1; Leukocyte Rolling; Lipopolysaccharides; Male; Mice; Mice, Inbred C57BL; Monocytes; NF-kappa B; Receptor, Cannabinoid, CB1; Receptor, Cannabinoid, CB2; rhoA GTP-Binding Protein; RNA, Messenger; Signal Transduction; Tumor Necrosis Factor-alpha; Vascular Cell Adhesion Molecule-1

Topics: Animals; Cannabinoid Receptor Agonists; Cannabinoid Receptor Modulators; Cannabinoids; Endocannabinoids; Inflammation; Ischemia; Ligands; Mice; Receptors, Cannabinoid; Reperfusion Injury

The effect of WIN55212-2, a cannabinoid receptor agonist, on acute inflammation of mouse ear was investigated. We found that topical application of WIN55212-2 suppressed ear swelling induced by 12-O-tetradecanoylphorbol 13-acetate or 2-arachidonoylglycerol. Similar inhibition was observed with CP55940, another cannabinoid receptor agonist, and HU-308, a cannabinoid CB(2) receptor-selective agonist. WIN55212-2 also suppressed the infiltration of leukocytes induced by 12-O-tetradecanoylphorbol 13-acetate. On the other hand, WIN55212-3, an inactive enantiomer of WIN55212-2, exerted only small effects on inflammation. Notably, SR144528, a cannabinoid CB(2) receptor antagonist, also suppressed inflammatory reactions in mouse ear. Thus, both the cannabinoid CB(2) receptor agonist and antagonist are capable of reducing inflammatory reactions. We then investigated the mechanism underlying WIN55212-2-induced suppression of inflammation using cultured cells. We found that the addition of WIN55212-2 together with 2-arachidonoylglycerol blocked 2-arachidonoylglycerol-induced migration of human promyelocytic leukemia HL-60 cells that had been differentiated into macrophage-like cells. The restoration of 2-arachidonoylglycerol-desensitized cells and WIN55212-2-desensitized cells from an anergic condition was examined next. We found that 2-arachidonoylglycerol-treated cells rapidly recovered the capacity to respond to 2-arachidonoylglycerol. On the other hand, the anergic condition toward 2-arachidonoylglycerol continued for a longer period after pretreatment with WIN55212-2. These results suggest that the anti-inflammatory activity of WIN55212-2 is attributable, at least in part, to interference with the actions of the endogenous ligand, 2-arachidonoylglycerol.

Topics: Administration, Cutaneous; Animals; Arachidonic Acids; Benzoxazines; Calcium; Cannabinoid Receptor Agonists; Cannabinoids; Cell Movement; Cyclohexanols; Dose-Response Relationship, Drug; Ear, External; Endocannabinoids; Glycerides; HL-60 Cells; Humans; Inflammation; Intracellular Fluid; Leukocytes; Ligands; Male; Mice; Mice, Inbred ICR; Morpholines; Naphthalenes; Peroxidase; Tetradecanoylphorbol Acetate

Involvement of cannabinoid CB(2) receptor and effect of cannabinoid CB(2) receptor antagonist/inverse agonists on cutaneous inflammation were investigated. Mice ears topically exposed to an ether-linked analogue of 2-arachidonoylglycerol (2-AG-E) or selective cannabinoid CB(2) receptor agonist, {4-[4-(1,1-dimethylheptyl)-2,6-dimethoxy-phenyl]-6.6-dimethyl-bicyclo[3.1.1]hept-2-en-2-yl}-methanol (HU-308), had early and late ear swelling (0--24 h and 1--8 days after exposure, respectively). Both types of responses induced by 2-AG-E were significantly suppressed by oral administration of cannabinoid CB(2) receptor antagonist/inverse agonists, [N-(benzo[1,3]dioxol-5-ylmethyl)-7-methoxy-2-oxo-8-pentyloxy-1,2-dihydroquinoline-3-carboxamide] (JTE-907) and {N-[(1S)-endo-1,3,3-trimethylbicyclo[2.2.1]heptan-2 yl]5-(4-chloro-3-methyl-phenyl)-1-(4-methylbenzyl)pyrazole-3-carboxamide}} (SR 144528). In contrast, JTE-907 did not affect arachidonic acid-induced swelling. Orally administered JTE-907 (0.1-10 mg/kg) and SR 144528 (1 mg/kg) also produced significant inhibition of dinitrofluorobenzene-induced ear swelling, with increased cannabinoid CB(2) receptor mRNA expression observed in the inflamed ear. These results suggest that cannabinoid CB(2) receptor is partially involved in local inflammatory responses and cannabinoid CB(2) receptor antagonist/inverse agonist has beneficial effects on ear swelling.

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Arachidonic Acid; Area Under Curve; Camphanes; Cannabinoids; Dinitrofluorobenzene; Dioxoles; Disease Models, Animal; Drug Eruptions; Ear, External; Female; Indomethacin; Inflammation; Lymph Nodes; Mice; Mice, Inbred BALB C; Pyrazoles; Quinolones; Receptor, Cannabinoid, CB2; RNA, Messenger; Time Factors