Journal: Antioxidants

Article Title: The Effect of Sea Buckthorn ( Hippophae rhamnoides L.) Seed Oil on UV-Induced Changes in Lipid Metabolism of Human Skin Cells

doi: 10.3390/antiox7090110

Figure Lengend Snippet: The level of endocannabinoids (AEA, 2-AG) and endocannabinoid receptors (CB1, CB2) in keratinocytes and fibroblasts after exposure to UVA (30 J/cm 2 and 20 J/cm 2 ) and UVB radiation (60 mJ/cm 2 and 200 mJ/cm 2 , respectively) and sea buckthorn seeds oil (500 ng/mL) treatment. Mean values ± SD of five independent experiments are presented. x statistically significant differences vs. control group, p < 0.05; y statistically significant differences between UVA/UVB and oil treated groups vs. oil treated control group, p < 0.05; a statistically significant differences between UVA and oil treated group vs. UVA treated group, p < 0.05; b statistically significant differences between UVB and oil treated group vs. UVB treated group, p < 0.05.

Article Snippet: Primary monoclonal antibodies against CB1 and CB2 (Santa Cruse Biotechnology, Santa Cruz, CA, USA) were used at a concentration of 1:1000.

Techniques: Control

Journal: International Journal of Molecular Sciences

Article Title: CB2 Receptor Activation Inhibits Melanoma Cell Transmigration through the Blood-Brain Barrier

doi: 10.3390/ijms15058063

Figure Lengend Snippet: Expression of cannabinoid and cannabinoid-like receptors in brain endothelial and melanoma cells. RT-PCR was performed to determine the expression of CB2A and CB2B (positive control: HL-60) transcriptional variants of human CB2 receptor in hCMEC/D3 brain endothelial and A2058 melanoma cells ( a , b ); the expression of transcriptional variant 1 and 2 of rat CB2 receptor ( c , d ) in rat brain endothelial cells (RBECs) (positive control: rat spleen), the expression of CB1 receptor ( e ); transcriptional variant 1 and 2 of GPR18 ( f , g ); GPR119 ( h ) and GPR55 ( i ) in hCMEC/D3 human brain endothelial cells and A2058 melanoma cells. Dotted arrows indicate the absence of specific bands.

Article Snippet: The selective CB2 agonist JWH-133 solution (diluted in Tocrisolve) was purchased from Tocris.

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Positive Control, Variant Assay

Journal: International Journal of Molecular Sciences

Article Title: CB2 Receptor Activation Inhibits Melanoma Cell Transmigration through the Blood-Brain Barrier

doi: 10.3390/ijms15058063

Figure Lengend Snippet: Effect of CB2 activation on the attachment of melanoma cells on the brain endothelium. Results are represented as % control ( i.e. , D3 + A2058) and given as mean ± SD. N = 3. * p < 0.05 as assessed by ANOVA and Bonferroni’s post-hoc test. ( a ) D3(jwh-133) and A2058(jwh-133) represent cells pre-treated with 10 μM JWH-133 for 4 h. D3 + A2058 + JWH-133 denotes cells treated with 10 μM JWH-133 during the 90 min adhesion assay; ( b ) JWH-133 (10 μM), U0126 (10 μM) and PTX (100 ng/mL) were applied in pre-treatment of both cell types and treatment during the adhesion assay.

Article Snippet: The selective CB2 agonist JWH-133 solution (diluted in Tocrisolve) was purchased from Tocris.

Techniques: Activation Assay, Control, Cell Adhesion Assay

Journal: International Journal of Molecular Sciences

Article Title: CB2 Receptor Activation Inhibits Melanoma Cell Transmigration through the Blood-Brain Barrier

doi: 10.3390/ijms15058063

Figure Lengend Snippet: Effect of CB2 activation on the transendothelial migration of melanoma cells. Results are represented as % control ( i.e. , RBEC + A2058) and given as mean ± SD. N = 3. * p < 0.01 (compared to control) as assessed by ANOVA and Bonferroni’s post-hoc test. ( a ) RBEC(jwh-133) represents endothelial cells pre-treated with 10 μM JWH-133 for 4 h. RBEC(jwh-133) + A2058(jwh-133) + JWH-133 denotes that both endothelial and melanoma cells were pre-treated with 10 μM JWH-133 for 4 h and treated with 10 μM JWH-133 during the 5 h transmigration assay; ( b ) JWH-133 (10 μM) and SR-144528 (10 μM) were applied in pre-treatment of both cell types and treatment during the transmigration assay.

Article Snippet: The selective CB2 agonist JWH-133 solution (diluted in Tocrisolve) was purchased from Tocris.

Techniques: Activation Assay, Migration, Control, Transmigration Assay

Journal: Arthritis Research & Therapy

Article Title: Elevated serum immunoglobulin level predicts high risk of 1-year recurrence in patients with Takayasu arteritis

doi: 10.1186/s13075-023-03016-8

Figure Lengend Snippet: CD138 expressions in the aortic wall of TAK and control group. A CD138 staining in the aortic wall of TAK patients and control group (scale bars, 100 μm; n = 3 in control group, n = 4 in TAK group). B The quantification of CD138 staining in the two groups ( n = 3 in control group, n = 4 in TAK group)

Article Snippet: Then, they were stained with primary antibody for CD138 (Item No.ab46506, Abcam, Cambridge, CB2 0AX, UK) overnight at 4 °C and secondary antibody for 0.5–1 h at room temperature and detected with 3,3′-diaminobenzidine.

Techniques: Staining

Journal: Mediators of Inflammation

Article Title: Cannabinoid Receptor CB2 Is Involved in Tetrahydrocannabinol-Induced Anti-Inflammation against Lipopolysaccharide in MG-63 Cells

doi: 10.1155/2015/362126

Figure Lengend Snippet: Experimental protocol diagram. (a) The MG-63 cells were assigned into five groups, including control, LPS, THC + LPS, AM630 + THC + LPS, and AM630 groups. After the treatments for 24 h, immunocytochemistry and western blot were taken to investigate the expression of CB2 receptor. (b) The MG-63 cells were assigned into seven groups as shown in this figure. After the treatments, ELISA was used to evaluate the inflammatory factors concentrations in the supernatants. (c) The MG-63 cells were assigned into five groups, including control, LPS, THC + LPS, CB2-siRNA + THC + LPS, and SC-siRNA + THC + AM630 groups. After the treatments, western blot was taken to investigate the expression of CB2 receptor and ELISA was used to assess the inflammatory factors release. (d) The cells were divided into control, LPS, THC + LPS, doxycycline (DOX) + THC + LPS, and DOX groups, western blot was taken to investigate the expression of cofilin-1 protein, and ELISA was taken to evaluate the concentrations of proinflammatory factors in the supernatants.

Article Snippet: CB2 antagonist AM630 and CB1 antagonist AM251 were obtained from Tocris Bioscience (UK).

Techniques: Control, Immunocytochemistry, Western Blot, Expressing, Enzyme-linked Immunosorbent Assay

Journal: Mediators of Inflammation

Article Title: Cannabinoid Receptor CB2 Is Involved in Tetrahydrocannabinol-Induced Anti-Inflammation against Lipopolysaccharide in MG-63 Cells

doi: 10.1155/2015/362126

Figure Lengend Snippet: THC upregulated CB2 receptor expression. Immunofluorescent staining (a) and western blot (b) were used to determine the CB2 protein expression. The cells were treated with or without 10 ng/mL LPS for 24 h in the presence or absence of 5 μ M THC or 10 μ M AM630 (CB2 receptor antagonist). The CB2 receptor expression (red) was observed by a confocal microscope, and stronger signal indicated higher expression of CB2 protein. Nuclei were counter-stained with DAPI (blue). Bar = 10 μ m. Results are means ± SD ( n = 6). * : P < 0.05; NS: no significance.

Article Snippet: CB2 antagonist AM630 and CB1 antagonist AM251 were obtained from Tocris Bioscience (UK).

Techniques: Expressing, Staining, Western Blot, Microscopy

Journal: Mediators of Inflammation

Article Title: Cannabinoid Receptor CB2 Is Involved in Tetrahydrocannabinol-Induced Anti-Inflammation against Lipopolysaccharide in MG-63 Cells

doi: 10.1155/2015/362126

Figure Lengend Snippet: THC decreased the release of TNF- α , IL-1 β , IL-6, and IL-8 from LPS-stimulated MG-63 cells. TNF- α (a), IL-1 β (b), IL-6 (c), and IL-8 (d) release were determined by ELISA kits. THC (5 μ M) decreased 10 ng/mL LPS-induced release of TNF- α , IL-1 β , IL-6, and IL-8, and CB2 antagonist AM630 (10 μ M) reversed these effects. Results are means ± SD ( n = 6). * : P < 0.05; NS: no significance.

Article Snippet: CB2 antagonist AM630 and CB1 antagonist AM251 were obtained from Tocris Bioscience (UK).

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Mediators of Inflammation

Article Title: Cannabinoid Receptor CB2 Is Involved in Tetrahydrocannabinol-Induced Anti-Inflammation against Lipopolysaccharide in MG-63 Cells

doi: 10.1155/2015/362126

Figure Lengend Snippet: THC reduced NF- κ B expression via CB2 receptor in LPS-stimulated MG-63 cells. NF- κ B expression was assessed by western blot. THC reduced LPS-induced upregulation of NF- κ B expression, and CB2 antagonist AM630 (a), not CB1 antagonist AM251 (b), partially abolished this effect. Results are means ± SD, ( n = 6). * : P < 0.05; NS: no significance.

Article Snippet: CB2 antagonist AM630 and CB1 antagonist AM251 were obtained from Tocris Bioscience (UK).

Techniques: Expressing, Western Blot

Journal: Mediators of Inflammation

Article Title: Cannabinoid Receptor CB2 Is Involved in Tetrahydrocannabinol-Induced Anti-Inflammation against Lipopolysaccharide in MG-63 Cells

doi: 10.1155/2015/362126

Figure Lengend Snippet: CB2-siRNA reversed THC-induced decrease of inflammatory factors release. MG-63 cells were assigned into five groups, control (no LPS, THC, or siRNA): cultured in drug-free medium; LPS: cells exposed to 10 ng/mL LPS for 24 h; THC + LPS: cells exposed to 5 μ M THC plus 10 ng/mL LPS for 24 h; CB2-siRNA + THC + LPS: cells incubated with CB2-siRNA for 24 h and then exposed to 5 μ M THC plus 10 ng/mL LPS for 24 h; scrambled siRNA (SC-siRNA) + THC + LPS: cells incubated with SC-siRNA for 24 h and then exposed to 5 μ M THC plus 10 ng/mL LPS for 24 h. (a) CB2-siRNA significantly downregulated the expression of CB2 receptor, assessed by western blot ( n = 3). (b) CB2-siRNA significantly reversed THC-induced effect on NF- κ B expression ( n = 6). (c)–(f) CB2-siRNA significantly reversed THC-induced effects on the release of TNF- α , IL-1 β , IL-6, and IL-8 ( n = 6). Results are means ± SD. * : P < 0.05; NS: no significance.

Article Snippet: CB2 antagonist AM630 and CB1 antagonist AM251 were obtained from Tocris Bioscience (UK).

Techniques: Control, Cell Culture, Incubation, Expressing, Western Blot

Journal: Mediators of Inflammation

Article Title: Cannabinoid Receptor CB2 Is Involved in Tetrahydrocannabinol-Induced Anti-Inflammation against Lipopolysaccharide in MG-63 Cells

doi: 10.1155/2015/362126

Figure Lengend Snippet: The upregulation of cofilin-1 inhibited the THC-induced anti-inflammation. (a) CB2 antagonist reversed the THC-induced effect on cofilin-1 expression. (b) Doxycycline induced an overexpression of cofilin-1 in the MG-63 cells. (c) Doxycycline abolished THC-induced reduction of IL-6 release. (d) Doxycycline abolished the THC-induced reduction of TNF- α release. The cells were treated with different drugs for 24 h (LPS: 10 ng/mL; THC: 5 μ M; AM630: 10 μ M; doxycycline: 0.1 μ g/mL), western blot was performed to evaluate the cofilin-1 expression, and ELISA was used to test the concentrations of IL-6 and TNF- α in the supernatants. Results are means ± SD ( n = 6). * : P < 0.05.

Article Snippet: CB2 antagonist AM630 and CB1 antagonist AM251 were obtained from Tocris Bioscience (UK).

Techniques: Expressing, Over Expression, Western Blot, Enzyme-linked Immunosorbent Assay

Journal: Cardiovascular & hematological disorders drug targets

Article Title: The Endocannabinoid System and Heart Disease: The Role of Cannabinoid Receptor Type 2

doi: 10.2174/1871529X18666180206161457

Figure Lengend Snippet: Synthetic CB2 ligands.

Article Snippet: A second CB2 knockout mouse was generated by Deltagen (Cnr2 tm1Dgen ) and is available from Jackson Labs (Bar Harbor, ME).

Techniques:

Journal: Cardiovascular & hematological disorders drug targets

Article Title: The Endocannabinoid System and Heart Disease: The Role of Cannabinoid Receptor Type 2

doi: 10.2174/1871529X18666180206161457

Figure Lengend Snippet: In vivo and in vitro models of CB2 in atherosclerosis.

Article Snippet: A second CB2 knockout mouse was generated by Deltagen (Cnr2 tm1Dgen ) and is available from Jackson Labs (Bar Harbor, ME).

Techniques: In Vivo, In Vitro, Activation Assay

Journal: Cardiovascular & hematological disorders drug targets

Article Title: The Endocannabinoid System and Heart Disease: The Role of Cannabinoid Receptor Type 2

doi: 10.2174/1871529X18666180206161457

Figure Lengend Snippet: The role of CB2 in ischemia/reperfusion.

Article Snippet: A second CB2 knockout mouse was generated by Deltagen (Cnr2 tm1Dgen ) and is available from Jackson Labs (Bar Harbor, ME).

Techniques:

Journal:

Article Title: Cannabinoids Ameliorate Pain and Reduce Disease Pathology in Cerulein-Induced Acute Pancreatitis

doi: 10.1053/j.gastro.2007.02.035

Figure Lengend Snippet: Induction of the endocannabinoid system in acute pancreatitis. Immunohistochemical detection of CB1 and CB2 is shown in human pancreas (A–C and G–I) or mouse pancreas (D–F and J–L) derived either from control subjects (A, D, G, and J) or subjects with acute pancreatitis (B and C, E and F, H and I, K and L). (C, F, I and L) Areas of necrosis within human and mouse acute pancreatitis are indicated by dotted lines. In normal human pancreas (A) CB1-immunoreactivity in acinar cells (▶; magnified in inset C), ducts (→), and nerves (magnified in insets), was weak but strong in acute pancreatitis (B, C). (D–F) Similarly, pancreas demonstrated increased immunoreactivity for CB1 after induction of acute pancreatitis with cerulein (E, F) over normal mouse pancreas (D). Intense CB1 immunostaining is observed on acinar cell necrosis (dotted arrow; F). (G–I) Moderate staining for CB2 is seen in normal human pancreas (G) with a slight increase in acute pancreatitis (H, I) in acinar cells (▶; magnified in inset I), ducts (→), and nerves (magnified in insets). Upon induction of acute pancreatitis in mice, there is a pronounced increase in CB2 immunoreactivity over expression levels in normal mouse pancreas (J–L). This is particularly present within acinar cell necrosis (dotted arrow; L). Original magnification: 40x (A and B, D and E, G and H, J and K) or 80x (F and L; insets in A–C and G–I) objective. (M, N) Immunoblot analysis of pancreas samples derived from acute pancreatitis (lanes 1–3) and control human donors (lanes 4–6) with antibodies recognizing CB1 (M) or CB2 (N). (▶) indicates anti-CB1 (at approximately 128 kDa) and anti-CB2-immunoreactive bands (at 38 and 26 kDa), which were abolished by preadsorption with the respective blocking peptide (lower lanes). Anti-ERK2 was used as an equal loading control. (O) In humans, pancreatic concentrations of the endocannabinoid anandamide (AEA) are higher in acute pancreatitis (black bars) than in normal pancreas (white bars; P < .05), whereas levels of combined 1- plus 2-arachidonoylglycerol (1 - AG + 2 - AG) are unchanged.

Article Snippet: Immunohistochemistry on paraffin sections of human pancreas was performed using rabbit anti-CB1 and anti-CB2 antibodies (Cayman Chemical, Ann Arbor, MI) as described previously.

Techniques: Immunohistochemical staining, Derivative Assay, Control, Immunostaining, Staining, Over Expression, Western Blot, Blocking Assay

Journal:

Article Title: Cannabinoids Ameliorate Pain and Reduce Disease Pathology in Cerulein-Induced Acute Pancreatitis

doi: 10.1053/j.gastro.2007.02.035

Figure Lengend Snippet: Levels of serological, molecular, and inflammatory markers in acute pancreatitis and effects of CB1/CB2 antagonists. (A) Levels of serum lipase, pancreatic MPO, and the pancreatic wet weight/dry weight ratio are increased in mice following cerulein-induced pancreatitis (black bars) over control animals (white bars; * P < .05). (B) Pancreatic levels of IL-6 protein and KC1 mRNA (normalized to GAPDH expression levels) rise significantly following induction of acute pancreatitis. Treatment with cannabinoid receptor antagonists AM251 or AM630 either alone or in combination did not change levels of these parameters, except pancreatic KC1 mRNA expression (P < .05). * and # represent P < .05 as compared with the saline group or the cerulein group, respectively. (*) Mann–Whitney U test. (#) Analysis of variance followed by post hoc Bonferroni’s multiple comparison test. n.s. = statistically not significant.

Article Snippet: Immunohistochemistry on paraffin sections of human pancreas was performed using rabbit anti-CB1 and anti-CB2 antibodies (Cayman Chemical, Ann Arbor, MI) as described previously.

Techniques: Control, Expressing, Saline, MANN-WHITNEY, Comparison