Journal: Annals of Translational Medicine

Article Title: Selective activation of cannabinoid receptor 2 regulates Treg/Th17 balance to ameliorate neutrophilic asthma in mice

doi: 10.21037/atm-21-2778

Figure Lengend Snippet: Effect of β-Car on pulmonary inflammation in neutrophilic asthma. Bronchial asthma was induced in mice by tobacco smoke exposure and ovalbumin stimulation (n=10), and subsequently administered β-Car, SR144528 or CMD178. SR144528 is a β-Car antagonist and CMD178 is a Treg inhibitor. (A) Lung tissue samples of the asthmatic mice were stained with hematoxylin & eosin and observed under a microscope (40×); (B,C,D) levels of inflammatory cytokines (TNF-α, IL-6, IL-8) assessed by ELISA; (E,F) levels of the cytokines IL-10 and TGF-β secreted by Treg as assessed by ELISA; (G,H) levels of the cytokines IL-17A and IL-22 secreted by Th17 as assessed by ELISA. ***P<0.001 vs. Control group; #P<0.05, ###P<0.001 vs. Asthma group; ^P<0.05, ^^P<0.01, ^^^P<0.001 vs. Asthma + β-Car group. β-Car, β-caryophyllene; ELISA, enzyme-linked immunosorbent assay; IL, interleukin; TGF, transforming growth factor; TNF, tumor necrosis factor; Treg, regulatory T cell.

Article Snippet: Reagents The Ovalbumin (OVA) (90%, DH015-4) was obtained from Guangzhou Dingguo Biotechnology Co., Ltd. β-Caryophyllene (β-Car; HY-N1415), SR144528 (HY-13439), and CMD178 (HY-P1453) were purchased from MedChemExpress (Monmouth Junction, NJ, USA).

Techniques: Staining, Microscopy, Enzyme-linked Immunosorbent Assay, Control

Journal: Annals of Translational Medicine

Article Title: Selective activation of cannabinoid receptor 2 regulates Treg/Th17 balance to ameliorate neutrophilic asthma in mice

doi: 10.21037/atm-21-2778

Figure Lengend Snippet: Effect of β-Car administration on levels of lymphocytes, eosinophils, and neutrophils in mice

Article Snippet: Reagents The Ovalbumin (OVA) (90%, DH015-4) was obtained from Guangzhou Dingguo Biotechnology Co., Ltd. β-Caryophyllene (β-Car; HY-N1415), SR144528 (HY-13439), and CMD178 (HY-P1453) were purchased from MedChemExpress (Monmouth Junction, NJ, USA).

Techniques: Control

Journal: Annals of Translational Medicine

Article Title: Selective activation of cannabinoid receptor 2 regulates Treg/Th17 balance to ameliorate neutrophilic asthma in mice

doi: 10.21037/atm-21-2778

Figure Lengend Snippet: Expression of CNR2 in lung tissue was induced by β-Car. Expression of CNR2 (100× and 400×) in right lung tissue from (A) Control group, (B) Asthma group, (C) Asthma + β-Car group, (D) Asthma + β-Car + SR144528 group and (E) Asthma + β-Car + CMD178 group was showed by immunohistochemical staining. β-Car, β-caryophyllene; CNR2, cannabinoid receptor 2.

Article Snippet: Reagents The Ovalbumin (OVA) (90%, DH015-4) was obtained from Guangzhou Dingguo Biotechnology Co., Ltd. β-Caryophyllene (β-Car; HY-N1415), SR144528 (HY-13439), and CMD178 (HY-P1453) were purchased from MedChemExpress (Monmouth Junction, NJ, USA).

Techniques: Expressing, Control, Immunohistochemical staining, Staining

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 inverse agonist SR-144528 (dissolved in ethanol) [ , ] was from Santa Cruz, the MEK1/2 inhibitor U0126 was from Cell Signaling and the Gi/Goα inhibitor pertussis toxin (PTX) was from Sigma-Aldrich (Budapest, Hungary).

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 inverse agonist SR-144528 (dissolved in ethanol) [ , ] was from Santa Cruz, the MEK1/2 inhibitor U0126 was from Cell Signaling and the Gi/Goα inhibitor pertussis toxin (PTX) was from Sigma-Aldrich (Budapest, Hungary).

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 inverse agonist SR-144528 (dissolved in ethanol) [ , ] was from Santa Cruz, the MEK1/2 inhibitor U0126 was from Cell Signaling and the Gi/Goα inhibitor pertussis toxin (PTX) was from Sigma-Aldrich (Budapest, Hungary).

Techniques: Activation Assay, Migration, Control, Transmigration Assay

Journal:

Article Title: Inhibition of guinea-pig and human sensory nerve activity and the cough reflex in guinea-pigs by cannabinoid (CB 2 ) receptor activation

doi: 10.1038/sj.bjp.0705435

Figure Lengend Snippet: JWH 133-mediated inhibitory responses on nerve depolarisations are SR144528 sensitive. The inhibitory action of JWH 133 on guinea-pig vagus nerve depolarisations induced by (a) hypertonic saline (2%), (b) capsaicin (1 μM) and (c) PGE2 (1 μM) is abolished in the presence of the CB2 receptor antagonist SR144528, and is unaffected by the CB1 receptor antagonist SR141716A. Nerve depolarisation responses were expressed as absolute values in mV depolarisation before and after drug additions, and then expressed a percentage change. The data were subjected to a paired two-tailed t-test, since the response to a stimulant was measured before and after drug treatment within the same nerve. **P<0.01 denotes the statistical significance compared to control responses in the same tissue prior to drug treatment and ###P<0.001 denotes the statistical significance between two different treatment groups using an unpaired t-test. Values are presented as the mean±s.e.m. percentage change of n=4 determinations.

Article Snippet: SR141716A and SR144528 were kind gifts from Novartis Institute, London, U.K. Cannabinoid agonists were obtained from Tocris Cookson Ltd (Bristol, U.K.).

Techniques: Saline, Two Tailed Test, Control

Journal:

Article Title: Inhibition of guinea-pig and human sensory nerve activity and the cough reflex in guinea-pigs by cannabinoid (CB 2 ) receptor activation

doi: 10.1038/sj.bjp.0705435

Figure Lengend Snippet: Effect of cannabinoid ligands on nerve depolarisations of isolated human vagus. Traces showing (a) the inhibitory effect of JWH 133 on nerve depolarisations induced by capsaicin, and (b) the blockade of this response by the CB2 receptor antagonist SR144528 from human vagus nerve.

Article Snippet: SR141716A and SR144528 were kind gifts from Novartis Institute, London, U.K. Cannabinoid agonists were obtained from Tocris Cookson Ltd (Bristol, U.K.).

Techniques: Isolation

Journal: Cell Death & Disease

Article Title: Celastrol alleviates renal fibrosis by upregulating cannabinoid receptor 2 expression

doi: 10.1038/s41419-018-0666-y

Figure Lengend Snippet: a – d Mice were treated with celastrol (1 mg/kg) by intraperitoneal injection immediately after UUO surgery. The celastrol+SR144528 group was additionally pretreated with SR144528 (1 mg/kg) 1 h before the celastrol injection. Both celastrol and SR144528 were administered daily. Mice were killed 7 days after UUO and kidney tissues were collected. a Representative micrographs show kidney injury using PAS staining and quantification of tubular injury scoring in different groups of mice. Bar = 100 μm. b Masson’s trichrome staining of kidney tissues sections. Bar = 50 μm. c Representative micrographs of the expression and distribution of α-SMA in kidney tissues using immunohistochemical staining. Bar = 100 μm. d Western blot analyses of renal fibronectin, collagen I, and α-SMA protein in kidney tissues. Representative western blot and quantitative data were presented. e Serum-starved HK-2 cells were pretreated with or without celastrol (500 nM) for 1 h and then stimulated with TGF-β1 (10 ng/ml) for 24 h. The celastrol+SR144528 group was additionally pretreated with SR144528 (1 μM) 30 min before the celastrol administration. Representative western blot analysis and quantification of fibronectin, collagen I, and α-SMA protein expression in HK-2 cells were presented. The medium group was used as the control for TGF-β1 treatment. Results are representative of three replicate experiments. All values are represented as mean ± SEM. n = 5 /group. * P < 0.05, ** P < 0.01, and *** P < 0.001

Article Snippet: The SR144528 and celastrol+SR144528 groups were pretreated with SR144528 (1 mg/kg, Adooq Bioscience, Irvine, CA, USA) 1 h before the surgery.

Techniques: Injection, Staining, Expressing, Immunohistochemical staining, Western Blot, Control