adenosine receptors  (Angelman Syndrome Foundation)

Journal: Bioactive Materials

Article Title: Immunomodulatory supramolecular hydrogel for rheumatoid arthritis management via adenosine A2A receptor-mediated macrophage remodeling

doi: 10.1016/j.bioactmat.2025.11.031

Figure Lengend Snippet: The design of an inflammatory macrophages-targeted, acid-sensitive PNSH for delivery MTX and its role in promoting A2AR activation on M2 macrophages for arthritis therapy. (A)The structure of acid-sensitive polymer-nanomedicine supramolecular hydrogels, composed of drug-loaded MTX NPs. (B) Schematic illustration of arthritis therapy targeting inflammatory joint network via mPECN NPs-mediated release of MTX. This approach leverages MTX targeting the A2AR and repolarization of macrophages from the pro-inflammatory M1 phenotype to the anti-inflammatory M2 phenotype. Activation of A2AR and macrophage repolarization by mPECN-MTX NPs synergistically enhanced the anti-inflammatory effect of PNSH-mediated nanomedicine in an arthritis rat model.

Article Snippet: The selective adenosine A2A receptor (A2AR) antagonist SCH58261 (HY-19533) was purchased from MedChemExpress (Shanghai, China).

Techniques: Activation Assay, Polymer

Journal: Bioactive Materials

Article Title: Immunomodulatory supramolecular hydrogel for rheumatoid arthritis management via adenosine A2A receptor-mediated macrophage remodeling

doi: 10.1016/j.bioactmat.2025.11.031

Figure Lengend Snippet: MTX@PNSH promotes anti-inflammatory M2 macrophages through the A2AR signaling pathway. (A) The intracellular cAMP levels in Raw 264.7 cells were determined by homogeneous time-resolved fluorescence (HTRF) following different treatments. (B) qRT-PCR analysis of relative mRNA of A2ar with different treatments. (C) Immunostaining of A2AR expression in Raw 264.7 cells with different treatments. (D) Flow cytometry analysis of A2AR + cells in Raw 264.7 cells with different treatments and the percentage of A2AR + cells in Raw 264.7 cells with different treatments. (E) qRT-PCR analysis of relative mRNA expression of Pdl1 without LPS treatment. (F) qRT-PCR analysis of relative mRNA expression of Ido1 without LPS treatment. (G) qRT-PCR analysis of relative mRNA expression of Pdl1 with LPS treatment. (H) qRT-PCR analysis of relative mRNA expression of Ido1 with LPS treatments. (I) Western blotting analysis of A2AR protein level in macrophages after 24 h of different treatment with or without A2AR inhibitor. (J) Quantitative analysis of A2AR protein level in different treatments with or without A2AR inhibitor. (K) Flow cytometry analysis of CD11b and CD39 in Raw 264.7 cells with different treatments and the percentage of CD11b + CD39 + macrophages. (L) Flow cytometry analysis of CD11b and CD73 in Raw 264.7 cells with different treatments and the percentage of CD11b + CD73 + macrophages. (M) Flow cytometry analysis of CD11b and PD-L1 in Raw 264.7 cells after 24 h of different treatments and the percentage of CD11b + PD-L1 + macrophages in Raw 264.7 cells after 24 h of different treatments. Data are presented as mean ± SD, n = 3. Statistical significance was determined using one-way ANOVA, followed by Dunnett's post hoc test for comparisons between groups.

Article Snippet: The selective adenosine A2A receptor (A2AR) antagonist SCH58261 (HY-19533) was purchased from MedChemExpress (Shanghai, China).

Techniques: Fluorescence, Quantitative RT-PCR, Immunostaining, Expressing, Flow Cytometry, Western Blot

Journal: Metabolites

Article Title: A 2A Receptor Activation Restores Lipid and Mitochondrial Homeostasis, Limiting Mycobacterium leprae Persistence in Human Monocytes

doi: 10.3390/metabo16050304

Figure Lengend Snippet: M. leprae infection selectively modulates ADO receptor expression in monocytes. Enriched human monocytes isolated from PBMCs were infected with M. leprae (MOI 10:1) for 48 h. The expression of A 1 , A 2B , A 3, and A 2A was assessed by flow cytometry and fluorescence microscopy. ( A – D ) Representative histograms showing ( A ) A 1 R, ( B ) A 2B R, ( C ) A 2A R, and ( D ) A 3 R expression in unstained cells (red), non-infected (NI; blue), and M. leprae -infected (orange) monocytes. ( E ) Representative fluorescence microscopy images of non-infected (NI) and M. leprae -infected human monocytes. A total of 200 cells per condition were stained with DAPI (blue) and an anti-A 2A antibody (green). ( F ) Bar graphs represent quantification of median fluorescence intensity (MFI), normalized to NI controls. Data are presented as mean ± SD from four independent donors, with each measurement conducted in duplicate. Scale bar = 10 μm. Statistical significance was determined using a paired t -test (* p < 0.05, ** p < 0.01).

Article Snippet: Cells were then incubated overnight at 4 °C with the following primary antibodies diluted 1:300 in blocking solution: rabbit anti-A 2A R antibody (Alomone Labs Cat# AAR-002; RRID: AB_2039707), and rabbit anti-ENT1 antibody (Abcam Cat# ab182023; RRID: AB_2732846).

Techniques: Infection, Expressing, Isolation, Flow Cytometry, Fluorescence, Microscopy, Staining

Journal: Metabolites

Article Title: A 2A Receptor Activation Restores Lipid and Mitochondrial Homeostasis, Limiting Mycobacterium leprae Persistence in Human Monocytes

doi: 10.3390/metabo16050304

Figure Lengend Snippet: Activation of the A 2A R decreases lipid droplet formation in M. leprae -infected human monocytes. Enriched human monocytes isolated from PBMCs were pretreated and infected with M. leprae . ( A ) Representative fluorescence microscopy images of non-infected (NI) and M. leprae -infected human monocytes treated with CGS21680 (100 µM) and or ZM241385 (50 nM). LDs were stained with Oil Red O (red), and the LD area per cell was analyzed in a total of 200 cells per condition. Nuclei were stained with DAPI (blue). White arrowheads indicate LDs. Scale bar = 10 μm. ( B ) LD quantification was performed using ImageJ software. NI = not infected. Data were presented as mean ± SD from at least three independent experiments conducted in technical duplicate. Statistical significance was determined by paired one-way ANOVA followed by Bonferroni’s multiple comparisons test (* p < 0.05).

Article Snippet: Cells were then incubated overnight at 4 °C with the following primary antibodies diluted 1:300 in blocking solution: rabbit anti-A 2A R antibody (Alomone Labs Cat# AAR-002; RRID: AB_2039707), and rabbit anti-ENT1 antibody (Abcam Cat# ab182023; RRID: AB_2732846).

Techniques: Activation Assay, Infection, Isolation, Fluorescence, Microscopy, Staining, Software

Journal: Metabolites

Article Title: A 2A Receptor Activation Restores Lipid and Mitochondrial Homeostasis, Limiting Mycobacterium leprae Persistence in Human Monocytes

doi: 10.3390/metabo16050304

Figure Lengend Snippet: A 2A R activation reduces M. leprae viability in human monocytes. Enriched human monocytes isolated from PBMCs were infected with M. leprae (MOI 10:1) for 48 h, and pretreated or not with the selective A 2A R agonist CGS21680 (50 nM or 100 µM) and/or the antagonist ZM241385 (50 nM). M. leprae viability was assessed by RT-qPCR using the ratio of 16S cDNA to 16S DNA represented by percentage relative to the control. ( A , B ) Pretreatment with CGS21680 significantly reduced M. leprae viability, and this effect was reversed by co-treatment with ZM241385. ( C ) Treatment with ZM241385 alone increased the bacterial survival compared to untreated infected cells. Data are presented as mean ± SD relative to the control, from at least four independent experiments in technical duplicate. Statistical significance was determined using one-way ANOVA followed by Bonferroni’s multiple comparisons test or using a paired t -test (* p < 0.05; ** p < 0.01; *** p < 0.001).

Article Snippet: Cells were then incubated overnight at 4 °C with the following primary antibodies diluted 1:300 in blocking solution: rabbit anti-A 2A R antibody (Alomone Labs Cat# AAR-002; RRID: AB_2039707), and rabbit anti-ENT1 antibody (Abcam Cat# ab182023; RRID: AB_2732846).

Techniques: Activation Assay, Isolation, Infection, Quantitative RT-PCR, Control

Journal: Drug Design, Development and Therapy

Article Title: Cordycepin Ameliorates Dextran Sulfate Sodium-Induced Ulcerative Colitis in Mice by Inhibiting IL-6/IL-6R-Mediated p38 MAPK and NF-κB Activation Through Adenosine A 2A Receptor Stimulation

doi: 10.2147/DDDT.S575035

Figure Lengend Snippet: Cordycepin abates DSS-induced damage in NCM460 cells by activating adenosine receptor A 2A . ( A ) The cAMP levels in NCM460 cells under treatment with 2% DSS and/or 1 μmol/L and 10 μmol/L COR. ( B ) The cAMP levels in NCM460 cells under treatment with 2% DSS, 10 μmol/L COR, 5 μmol/L SCH58261 (SCH), and / or 5 μmol/L DPCPX for 24 h. The release levels of ( C ) LDH, ( D ) IL-1β, ( E ) IL-6, and ( F ) TNF-α were measured by using Elisa. ( G ) Representative images of PI staining and the percentage of PI-positive cells, scar bar = 100 μm. ( H – J ) The protein expression of ZO-1, A 2A AR and A 1 AR in NCM460 cells was determined by using Western blotting assay. The results were representative of three independent experiments and expressed as mean ± SEM. Data were compared using two-tailed Student’s t tests. vs. control group; # P < 0.05, vs. control group; *P < 0.05, **P < 0.01 and ***P < 0.001 vs. the 2% DSS group.

Article Snippet: Adenosine A 2A receptor Antibody (A 2A AR, sc-32261) was purchased from Santa Cruz Biotechnology (USA).

Techniques: Enzyme-linked Immunosorbent Assay, Staining, Expressing, Western Blot, Two Tailed Test, Control

Journal: Drug Design, Development and Therapy

Article Title: Cordycepin Ameliorates Dextran Sulfate Sodium-Induced Ulcerative Colitis in Mice by Inhibiting IL-6/IL-6R-Mediated p38 MAPK and NF-κB Activation Through Adenosine A 2A Receptor Stimulation

doi: 10.2147/DDDT.S575035

Figure Lengend Snippet: The results of molecular docking and molecular dynamics simulation of adenosine receptor A 2A -cordycepin. ( A ) 2D (left) and 3D (right) molecular docking diagrams of A 2A AR-COR. The PDB ID of the A 2A AR protein is 5nm4. The yellow stick represents the structure of COR, and the cartoon depicts the protein structure. The green sticks indicate amino acid residues within 4 Å that form hydrogen bonds with COR. Yellow dashed lines represent hydrogen bonds, with adjacent numbers denoting the distance between COR and the hydrogen-bonded amino acids. White sticks represent amino acids involved in hydrophobic interactions with COR. ( B ) The Root Mean Square Deviation (RMSD). ( C ) The number of hydrogen bonds of the complex. ( D ) The Root Mean Square Fluctuation (RMSF) values for the complex. ( E ) The Radius of Gyration (Rg) values of the complex. ( F ) Solvent Accessible Surface Area (SASA) of the complex.

Article Snippet: Adenosine A 2A receptor Antibody (A 2A AR, sc-32261) was purchased from Santa Cruz Biotechnology (USA).

Techniques: Solvent

Journal: Drug Design, Development and Therapy

Article Title: Cordycepin Ameliorates Dextran Sulfate Sodium-Induced Ulcerative Colitis in Mice by Inhibiting IL-6/IL-6R-Mediated p38 MAPK and NF-κB Activation Through Adenosine A 2A Receptor Stimulation

doi: 10.2147/DDDT.S575035

Figure Lengend Snippet: Cordycepin inhibits IL-6/IL-6R-mediated p38 MAPK and NF-κB phosphorylation activation relying on activation of adenosine receptor A 2A . ( A – E ) Representative blot bands and relative expression levels for IL-6R, NF-κB P-p65, NF-κB p65, NF-κB P-p65/p65, detected by Western blotting. ( F – I ) Representative blot bands and relative expression levels for P-p38 MAPK, p38 MAPK, P-p38/p38 MAPK. ( J – M ) Representative blot bands and relative expression levels for P-STAT3, STAT3, P-STAT3/STAT3. Data were presented as the means ± SEM of three mice in each group and were compared using two-tailed Student’s t tests. # P < 0.05, ## P < 0.01 vs. control group; *P < 0.05, **P < 0.01 vs. the DSS model (DSS) group.

Article Snippet: Adenosine A 2A receptor Antibody (A 2A AR, sc-32261) was purchased from Santa Cruz Biotechnology (USA).

Techniques: Phospho-proteomics, Activation Assay, Expressing, Western Blot, Two Tailed Test, Control

Journal: Drug Design, Development and Therapy

Article Title: Cordycepin Ameliorates Dextran Sulfate Sodium-Induced Ulcerative Colitis in Mice by Inhibiting IL-6/IL-6R-Mediated p38 MAPK and NF-κB Activation Through Adenosine A 2A Receptor Stimulation

doi: 10.2147/DDDT.S575035

Figure Lengend Snippet: The ameliorative effect of cordycepin on DSS-induced colitis is attributed to its activation of A 2A AR, which inhibits IL-6/IL-6R-mediated phosphorylation activation of p38 MAPK and NF-κB.

Article Snippet: Adenosine A 2A receptor Antibody (A 2A AR, sc-32261) was purchased from Santa Cruz Biotechnology (USA).

Techniques: Activation Assay, Phospho-proteomics