cb2r antagonist am630  (MedChemExpress)

Journal: Molecular Pain

Article Title: CB2 receptor agonist AM1241 regulating the polarization of microglia reduces morphine tolerance through IL-4/STAT6 pathway

doi: 10.1177/17448069251374281

Figure Lengend Snippet: Effects of AM1241 on markers of microglia polarization in vitro. There were six different groups in BV2 and HMC3 cells. Cells were treated with control, LPS, LPS+AM1241, LPS+AM281+AM1241, LPS+AM630+AM1241 in combination with morphine (Mor) or alone plus morphine (Mor). Relative iNOS mRNA (a, c) and SOCS3 mRNA (b, d) level adjusted to GAPDH. Western blot analysis of iNOS and SOCS3 (e, f). The iNOS protein (g, i) and SOCS3 (h, j) protein fold of control. One-way analysis of variance (ANOVA) followed by Bonferroni’s multiple comparisons test was used for analyzing the Western blot and qPCR in different group ( p < 0.05). The data in this study was shown as mean ± standard deviation (SD). p < 0.05 was considered statistically significant. *p < 0.05: the LPS+Mor group compared with the control group; $ p < 0.05: the different groups compared with the LPS+AM1241+Mor group.

Article Snippet: The CB2R agonist AM1241 was obtained from APExBIO, while CB2R antagonist AM630, CB1R antagonist AM281 and IL-4I were obtained from MedChemExpress.

Techniques: In Vitro, Control, Western Blot, Standard Deviation

Journal: Molecular Pain

Article Title: CB2 receptor agonist AM1241 regulating the polarization of microglia reduces morphine tolerance through IL-4/STAT6 pathway

doi: 10.1177/17448069251374281

Figure Lengend Snippet: Effects of AM1241 on the PWL and polarization of microglia in a morphine tolerance model. (a): mice were divided into five groups. Vehicle (VEH), AM1241, AM281+AM1241, AM630+AM1241 in combination with morphine (10 mg/kg, M10) or alone plus morphine was administered once daily for 7 days ( n = 6 in each group). Thermal withdrawal latency were measured 30 min after the last drug administration. Data are exhibited as mean ± SD. p < 0.05 was defined as statistically significant. * p < 0.05: different groups compared with VEH+SAL group; $ p < 0.05: different groups compared with the AM1241+M10; # p < 0.05: compared with the day 1 in corresponding group. (b): On day 8, morphine tolerance was determined by measuring PWL 30, 60, 90 and 120 min after 5 mg/kg morphine (M5) treatment. Two-way analysis of variance (ANOVA) followed by Bonferroni’s multiple comparisons test was used for analyzing the PWL. The spinal cord was harvested for qPCR and Western blot ( n = 3 per group) after the PWL on day 8. The mRNA and protein of microglia was determined as shown in C-G. Relative iNOS mRNA (c) and SOCS3 mRNA (d) level adjusted to GAPDH. Western blot analysis of iNOS and SOCS3 in (e). The iNOS protein (f), and SOCS3 protein (g) fold of control. The data were analyzed via one-way analysis of variance followed by Bonferroni’s multiple comparisons test. The mRNA and protein expression relative to the control ± SD ( n = 3 in each group) are presented. p < 0.05 was defined as statistically significant. * p < 0.05: different groups compared with the VEH+SAL+SAL group; $ p < 0.05: the different groups compared with the AM1241+M10+M5 group. The VEH+SAL+SAL group served as the control group.

Article Snippet: The CB2R agonist AM1241 was obtained from APExBIO, while CB2R antagonist AM630, CB1R antagonist AM281 and IL-4I were obtained from MedChemExpress.

Techniques: Western Blot, Control, Expressing

Journal: Journal of neuroinflammation

Article Title: Cannabinoids modulate the microbiota-gut-brain axis in HIV/SIV infection by reducing neuroinflammation and dysbiosis while concurrently elevating endocannabinoid and indole-3-propionate levels.

doi: 10.1186/s12974-023-02729-6

Figure Lengend Snippet: Fig. 4 THC can counteract the post-transcriptional silencing of WFS1 by miR-142-3p. The heat map shows all differentially expressed (p ≤ 0.05) miRNAs in the BG of VEH/SIV (A) and THC/SIV (B) relative to uninfected controls RMs and in VEH/SIV compared to THC/SIV RMs (C). MiRNA species originating from the opposite arm of the precursor are denoted with an asterisk (*). Red arrows (A) indicate inflammation-associated miRNAs differentially upregulated in BG of VEH/SIV RMs. Blue and green arrows indicate miRNAs commonly up and downregulated in BG of VEH/SIV (A) and THC/SIV (B) RMs, respectively. MiRNA–mRNA duplex showing a single miR-142-3p binding site on the RM WFS1 (D) mRNA 3’ UTR. RT-qPCR validation of miR-142-3p expression in BG of VEH/SIV relative to uninfected control RMs (E). Luciferase reporter vectors containing a single highly conserved miR-142-3p (F) binding site on the RM WFS1 mRNA 3′ UTR or the corresponding construct with the binding sites deleted (WFS1 DEL) were co-transfected into HEK293 cells with 30 nM miR-142-3p or negative control mimic. Firefly and Renilla luciferase activities were detected using the Dual-Glo luciferase assay system 96 h after transfection. Luciferase reporter assays were performed thrice in six replicate wells (F). Representative immunofluorescence images showing the expression of WFS1 (red) protein at 96 h post-transfection of HCN2 neuronal cells with 30 nM LNA-conjugated FAM-labeled negative control (G) or miR-142-3p (green) (H) mimics and images were quantitated (L). miR-142-3p transfected HCN2 cells were treated with DMSO (H), THC (I), AM251 + THC (J), AM630 + THC (K) 96 h post-transfection. Cells were fixed and stained after 18 h and the expression of WFS1 (red) protein and nuclear staining using DAPI (blue) were quantitated (L). Experiments were performed in triplicate wells using 3 µM of THC, 10 µM of AM251/AM630, and repeated thrice. Firefly/Renilla ratios and immunofluorescence data were analyzed using one-way ANOVA followed by Tukey’s multiple comparison post hoc test. A p-value of < 0.05 was considered significant

Article Snippet: After confirming protein expression of both CB1R and CB2R (Additional file 1: Fig. S3A, B), HCN2 cells were preincubated with 10 μM of the CB1R antagonist AM251 or the CB2R antagonist AM630 (Tocris Bioscience, Minneapolis, MN) for 1 h followed by 3 μM THC to identify the specific cannabinoid receptor (CB1R or CB2R) involved in transducing the effects of THC on WFS1 protein expression.

Techniques: Binding Assay, Quantitative RT-PCR, Biomarker Discovery, Expressing, Control, Luciferase, Construct, Transfection, Negative Control, Immunofluorescence, Labeling, Staining, Comparison