ccr2 inhibitor  (Tocris)

Journal: Bioactive Materials

Article Title: Spatiotemporally programming the immune-osteogenic cascade with a dual-immunomodulatory scaffold for functional bone regeneration

doi: 10.1016/j.bioactmat.2026.04.002

Figure Lengend Snippet: Immune regulation and endogenous bone regeneration mechanism investigation. A) Network diagram showing the number of interactions between six subclusters. B) KEGG enrichment analysis of the upregulated DEGs in DIBS group compared to the HA group. C) Circular visualization of related pathway–gene enrichment analysis. D) Heatmap of key gene regulation in specific pathways. E) qRT-PCR validation for key gene expression in specific pathways. F) The interaction networks showing the correlation of representative immunomodulatory genes (CCL2, CCL20, Sfrp1, and Stat3, etc.) with angiogenesis/osteogenesis and macrophage regulation gene sets. G) Flow cytometry analysis and quantification of CCR2 F4/80 macrophage in peripheral blood. H) Immunofluorescence staining analysis of macrophage polarization inside scaffolds (one week after intramuscular implantation). I) Macrophage proliferation assay in a CCR2-dependent manner. J and K) Macrophage polarization assay in a CCR2-dependent manner. Data are represented as means ± SD, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001; $ p < 0.05 (vs group without inhibitor), $$ p < 0.01 (vs group without inhibitor), $$$ p < 0.001 (vs group without inhibitor), $$$$ p < 0.0001 (vs group without inhibitor). ns, not significant.

Article Snippet: Throughout the experimental period, sustained CCR2 inhibition was achieved via daily intraperitoneal injections (2 mg/kg) of the highly selective CCR2 inhibitor RS504393 (Cat. No. HY-15418, MCE).

Techniques: Quantitative RT-PCR, Biomarker Discovery, Gene Expression, Flow Cytometry, Immunofluorescence, Staining, Proliferation Assay

Journal: Bioactive Materials

Article Title: Spatiotemporally programming the immune-osteogenic cascade with a dual-immunomodulatory scaffold for functional bone regeneration

doi: 10.1016/j.bioactmat.2026.04.002

Figure Lengend Snippet: Revascularization and osteogenesis are reinforced by M2 macrophage activation via the CCL2/CCR2 pathway. A and B) HUVECs and BMSCs proliferation assay under M2 macrophage activation. Created with BioRender.com . C) Migration assay and quantification of HUVECs. D) Tube formation assay and quantification of HUVECs. E and F) Early and later osteogenic differentiation of BMSC influenced by macrophage-induced microenvironment. Data are represented as means ± SD, ∗ p < 0.05 (vs Control), ∗∗ p < 0.01 (vs Control), ∗∗∗ p < 0.001 (vs Control), ∗∗∗∗ p < 0.0001 (vs Control); $ p < 0.05 (vs group without inhibitor), $$ p < 0.01 (vs group without inhibitor), $$$ p < 0.001 (vs group without inhibitor), $$$$ p < 0.0001 (vs group without inhibitor). ns, not significant.

Article Snippet: Throughout the experimental period, sustained CCR2 inhibition was achieved via daily intraperitoneal injections (2 mg/kg) of the highly selective CCR2 inhibitor RS504393 (Cat. No. HY-15418, MCE).

Techniques: Activation Assay, Proliferation Assay, Migration, Tube Formation Assay, Control

Journal: Journal of Advanced Research

Article Title: Novel function of macrophage migration inhibitory factor in regulating post-infarct inflammation and the therapeutic significance

doi: 10.1016/j.jare.2025.05.030

Figure Lengend Snippet: Macrophage migration inhibitory factor (MIF) promotes inflammatory cell migration and infiltration through C – C motif chemokine receptor 2 (CCR2) and C-X-C motif chemokine receptor (CXCR) 4 and promotes splenic monocyte mobilization via interaction with angiotensin Ⅱ type 1 receptor (AT-1R) after myocardial infarction (MI). A, peripheral blood mononuclear cells (PBMCs) chemotaxis in response to homogenized sham or infarct heart tissue from wild type (WT) mice at day 3 post-MI, or MI homogenate together with inhibition of C–C motif chemokine receptor 2 (CCR2), C-X-C motif chemokine receptor (CXCR) 2 and CXCR4, respectively. Cells/HPF, cells/high power field. PBMCs harvested from WT mice at 3 days post-MI. CCR2i, CCR2 inhibition. CXCR2i, CXCR2 inhibition. CXCR4i, CXCR4 inhibition. n = 4–8 per group. *** P < 0.001 vs. sham . B, PBMCs chemotaxis in response to recombinant human MIF (rMIF) or rMIF with inhibition of CCR2, CXCR2 and CXCR4, respectively. PBMCs harvested from WT mice at 3 days post-MI. n = 4–8 per group. *** P < 0.001 vs. control . C, Representative immunoblotting images for monocyte chemoattractant protein-1 (MCP-1), CXCR4 and internal reference protein (heat shock protein 60, HSP-60) in hearts from WT and global MIF deficient (MIFKO) mice with sham-operation or 72 h MI. D-E, Quantitative analysis of MCP-1 and CXCR4 expression. n = 6–9 per group. ** P < 0.01 vs. sham in the same genotype. *** P < 0.001 vs. sham in the same genotype. F, Splenic monocyte (from normal WT mice) chemotaxis in response to plasma from normal mice or mice with 1.5 h MI, or MI plasma together with losartan. Lor, losartan, AT-1R inhibitor. n = 6–8 per group. *** P < 0.001 vs. normal. G, Splenic monocytes (from normal WT mice) chemotaxis in response to rMIF and angiotensin II (Ang II), or together with losartan addition, respectively. n = 6–8 per group. ** P < 0.01 vs. Control. *** P < 0.001 vs. Control. H, Representative immunoblotting images for AT-1R and internal reference protein (HSP-60) in spleens from WT and global MIF deficient (MIFKO) mice with sham-operation and 1.5 h MI. I, Quantitative analysis of AT-1R expression. n = 8 per group. *** P < 0.001 vs. sham in the same genotype . J. Co-immunoprecipitation (Co-IP) assay using an anti-AT-1R antibody or IgG demonstrated the interaction between AT-1R and MIF, as well as between AT-1R and CD74 in splenic monocytes and spleen tissue. Input, input sample. IgG, negative control sample. IP, Co-IP sample.

Article Snippet: For both groups 1 and 2, PBMCs were pre-treated for 1 h using DMEM with or without CCR2 inhibitor (CCR2 antagonist 4 hydrochloride, 100 mM), CXCR2 inhibitor (SB225002, 800 nM) or CXCR4 inhibitor (Plerixafor, AMD3100, 10 μM) (all from MedChemExpress, USA).

Techniques: Migration, Chemotaxis Assay, Inhibition, Recombinant, Control, Western Blot, Expressing, Clinical Proteomics, Co-Immunoprecipitation Assay, Negative Control

Journal: Inflammation

Article Title: Activation of β2-Adrenergic Receptor Alleviates Viral Myocarditis by Regulating Energy Metabolism in Monocyte-derived Macrophages via the AMPK Pathway

doi: 10.1007/s10753-025-02382-6

Figure Lengend Snippet: Activating β2-AR attenuates inflammation and fibrosis in VMC. A - B Representative flow cytometry images (A) and statistical results (B) of β2-AR expression on CD45 + CD64 + F4/80 + CCR2 + macrophages in control and myocarditis model mice at 1 week and 2 weeks post-CVB3 infection ( n = 6/group). C - D Representative H&E-stained images of myocardial tissue from each group at 1 week (C) and 2 weeks (D) post-CVB3 infection (original magnification, 200 × ; scale bars = 50 µm). E – F Representative Masson's trichrome staining images from each group at 1 week (E) and 2 weeks (F) post-CVB3 infection (original magnification, 200 × ; scale bars = 50 µm). G - H Representative cardiac ultrasound images of the mice in each group at 1 week (G) and 2 weeks (H) post-CVB3 infection. (I) Statistical analysis of the myocardial pathological scores for each group at 1 week and 2 weeks post-CVB3 infection ( n = 6/group). J Statistical analysis of cardiac collagen volume fractions in each group at 1 week and 2 weeks post-CVB3 infection ( n = 6/group). K - L Statistical analysis of the LVFS (K) and LVEF (L) of each group at 1 week and 2 weeks post-CVB3 infection ( n = 6/group). M – O Real-time qPCR analysis of CCL2 (M), TNF-α (N), and IL-10 (O) expression in each group at 1 week and 2 weeks post-CVB3 infection ( n = 6/group). The data are presented as the means ± SEMs. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, ns = not significant. Abbreviations: CCL2, C–C motif chemokine ligand 2; CVB3, coxsackievirus B3; H&E, hematoxylin and eosin; LVFS, left ventricular fractional shortening; LVEF, left ventricular ejection fraction; IL-10, interleukin 10; qPCR, real-time quantitative polymerase chain reaction; SEM, standard error of the mean; TNF-α, tumor necrosis factor-α; VMC, viral myocarditis; β2-adrenergic receptor, β2-AR

Article Snippet: Following established protocols [ ], the C–C motif chemokine receptor 2 (CCR2) inhibitor INCB3344 (HY-50674, MCE, USA) was delivered intraperitoneally at 30 mg/kg/day to VMC mice, starting on Day 1 and continuing until Day 14 post-CVB3 infection.

Techniques: Flow Cytometry, Expressing, Control, Infection, Staining, Real-time Polymerase Chain Reaction

Journal: Inflammation

Article Title: Activation of β2-Adrenergic Receptor Alleviates Viral Myocarditis by Regulating Energy Metabolism in Monocyte-derived Macrophages via the AMPK Pathway

doi: 10.1007/s10753-025-02382-6

Figure Lengend Snippet: Effects of activating β2-AR on the frequencies and subsets of monocytes/macrophages. A Representative flow cytometry images of cardiac macrophage subsets among CD45 + CD64 + F4/80 + cells in the control, CVB3 + vehicle, and CVB3 + formoterol-treated groups at 1 week and 2 weeks post-infection. B - C Statistical analysis of CCR2 + macrophages (B) and CCR2 − macrophages (C) among cardiac macrophages in the control, CVB3 + vehicle, and CVB3 + formoterol-treated groups at 1 week and 2 weeks post-infection ( n = 6/group). D Representative flow cytometry images of Ly6C high and Ly6C low monocytes among blood CD45 + CD11b + Ly6G − cells in the control, CVB3 + vehicle, and CVB3 + formoterol-treated groups at 1 week and 2 weeks post-infection. E – F Statistical analysis of Ly6C high monocytes (E) and Ly6C low monocytes (F) among blood CD45 + CD11b + Ly6G − cells in the control, CVB3 + vehicle, and CVB3 + formoterol-treated groups at 1 week and 2 weeks post-infection ( n = 6/group). G - H Representative flow cytometry images (G) and statistical analysis (H) of Ly6C + monocytes among splenic CD45 + CD11b + Ly6G − cells in the control, CVB3 + vehicle, and CVB3 + formoterol-treated groups at 1 week and 2 weeks post-infection (n = 6/group). The data are presented as the means ± SEMs. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, ns = not significant. Abbreviations: CCR2, C-C chemokine receptor type 2; CVB3, coxsackievirus B3; SEM, standard error of the mean; β2-adrenergic receptor, β2-AR

Article Snippet: Following established protocols [ ], the C–C motif chemokine receptor 2 (CCR2) inhibitor INCB3344 (HY-50674, MCE, USA) was delivered intraperitoneally at 30 mg/kg/day to VMC mice, starting on Day 1 and continuing until Day 14 post-CVB3 infection.

Techniques: Flow Cytometry, Control, Infection

Journal: Inflammation

Article Title: Activation of β2-Adrenergic Receptor Alleviates Viral Myocarditis by Regulating Energy Metabolism in Monocyte-derived Macrophages via the AMPK Pathway

doi: 10.1007/s10753-025-02382-6

Figure Lengend Snippet: Effects of activating β2-AR on the polarization and function of cardiac CCR2 + macrophages and CCR2 − macrophages. A - B Representative flow cytometry images (A) and statistical analysis (B) of CCR2 + iNOS + M1 macrophages among CD45 + CD64 + F4/80 + CCR2 + macrophages in the control, CVB3 + vehicle, and CVB3 + formoterol groups at 1 week and 2 weeks post-infection ( n = 6/group). C - F Representative flow cytometry images and statistical analysis of TNF-α (C-D) and CCL2 (E–F) secreted by CD45 + CD64 + F4/80 + CCR2 + macrophages in the control, CVB3 + vehicle, and CVB3 + formoterol groups at 1 week and 2 weeks post-infection ( n = 6/group). G - H Representative flow cytometry images (G) and statistical analysis (H) of CCR2 + CD206 + M2 macrophages among CD45 + CD64 + F4/80 + CCR2 + macrophages in the control, CVB3 + vehicle, and CVB3 + formoterol groups at 1 week and 2 weeks post-infection ( n = 6/group). I - J Representative flow cytometry images (I) and statistical analysis (J) of IL-10 secretion by CD45 + CD64 + F4/80 + CCR2 + macrophages in the control, CVB3 + vehicle, and CVB3 + formoterol groups at 1 week and 2 weeks post-infection ( n = 6/group). K - L Representative flow cytometry images (K) and statistical analysis (L) of CCR2 − iNOS + M1 macrophages among CD45 + CD64 + F4/80 + CCR2 − macrophages in the control, CVB3 + vehicle, and CVB3 + formoterol groups at 1 week and 2 weeks post-infection ( n = 6/group). M - P Representative flow cytometry images and statistical analysis of TNF-α (M–N) and CCL2 (O-P) secretion by CD45 + CD64 + F4/80 + CCR2 − macrophages in the control, CVB3 + vehicle, and CVB3 + formoterol groups at 1 week and 2 weeks post-infection ( n = 6/group). Q - R Representative flow cytometry images (Q) and statistical analysis (R) of CCR2 − CD206 + M2 macrophages among CD45 + CD64 + F4/80 + CCR2 − macrophages in the control, CVB3 + vehicle, and CVB3 + formoterol groups at 1 week and 2 weeks post-infection ( n = 6/group). S - T Representative flow cytometry images (S) and statistical analysis (T) of IL-10 secretion by CD45 + CD64 + F4/80 + CCR2 − macrophages in the control, CVB3 + vehicle, and CVB3 + formoterol groups at 1 week and 2 weeks post-infection ( n = 6/group). The data are presented as the means ± SEMs. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, ns = not significant. Abbreviations: CCL2, C–C motif chemokine ligand 2; CCR2, C–C chemokine receptor type 2; CVB3, coxsackievirus B3; IL-10, interleukin 10; iNOS, inducible nitric oxide synthase; SEM, standard error of the mean; TNF-α, tumor necrosis factor-α; β2-adrenergic receptor, β2-AR

Article Snippet: Following established protocols [ ], the C–C motif chemokine receptor 2 (CCR2) inhibitor INCB3344 (HY-50674, MCE, USA) was delivered intraperitoneally at 30 mg/kg/day to VMC mice, starting on Day 1 and continuing until Day 14 post-CVB3 infection.

Techniques: Flow Cytometry, Control, Infection

Journal: Inflammation

Article Title: Activation of β2-Adrenergic Receptor Alleviates Viral Myocarditis by Regulating Energy Metabolism in Monocyte-derived Macrophages via the AMPK Pathway

doi: 10.1007/s10753-025-02382-6

Figure Lengend Snippet: Activating β2-AR attenuates inflammation and fibrosis in VMC via CCR2 + macrophages. A Schematic illustrating macrophage depletion using clodronate liposomes (Lipo-Clod). The mice received intraperitoneal injections of 200 µl clodronate liposomes or PBS-containing liposomes (Lipo-PBS) 2 days before CVB3 infection, followed by injections on days 5 and 10 post-infection. B - C Representative flow cytometry images (B) and statistical analysis (C) of CD11b + Ly6G − monocytes among splenic CD45 + cells in the Vehicle and Lipo-Clod groups at 1 week and 2 weeks post-CVB3 infection ( n = 6/group). D Schematic showing CCR2 + monocyte/macrophage inhibition via INCB3344 which was administered from day 1 to day 7 or day 14 post-CVB3 infection. E – F Representative flow cytometry images (E) and statistical analysis (F) of Ly6C high monocytes among blood CD45 + CD11b + Ly6G − cells in the Vehicle and INCB3344 groups at 1 week and 2 weeks post-CVB3 infection ( n = 6/group). G - H Representative H&E staining (G) and Masson’s trichrome staining (H) images of the myocardium from the Vehicle, Lipo-Clod, Formoterol, and Formoterol + Lipo-Clod groups at 1 week and 2 weeks post-CVB3 infection (original magnification, 200 × ; scale bars = 50 µm). I - J Statistical analysis of myocardial pathological scores (I) and cardiac collagen volume fractions (J) in each group at 1 week and 2 weeks post-CVB3 infection ( n = 6/group). K - L Statistical analysis of myocardial pathological scores (K) and cardiac collagen volume fractions (L) in the Vehicle, INCB3344, Formoterol, and Formoterol + INCB3344 groups 1 week and 2 weeks post-CVB3 infection ( n = 6/group). M - N Statistical analysis of myocardial pathological scores (M) and cardiac collagen volume fractions (N) in each group at 1 week and 2 weeks post-CVB3 infection ( n = 6/group). The data are presented as the means ± SEMs. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, ns = not significant. Abbreviations: CCR2, C-C chemokine receptor type 2; CVB3, coxsackievirus B3; VMC, viral myocarditis; SEM, standard error of the mean; ns, not statistically significant; β2-adrenergic receptor, β2-AR

Article Snippet: Following established protocols [ ], the C–C motif chemokine receptor 2 (CCR2) inhibitor INCB3344 (HY-50674, MCE, USA) was delivered intraperitoneally at 30 mg/kg/day to VMC mice, starting on Day 1 and continuing until Day 14 post-CVB3 infection.

Techniques: Liposomes, Infection, Flow Cytometry, Inhibition, Staining