ccr5 antagonist maraviroc  (MedChemExpress)

Journal: Advanced Science

Article Title: USP11‐PGAM5 Axis Promotes Neurotoxic Astrocyte Reactivity by Aggravating the mtDNA‐cGAS‐STING Pathway After Intracerebral Hemorrhage

doi: 10.1002/advs.202514283

Figure Lengend Snippet: Astrocytic PGAM5 mediates neutrophil TEM and NET release via the CCL5‐CCR5 chemotactic axis. A,B) Immunofluorescence staining of IBA1 and CD86, and statistical analysis of CD86+ cells in IBA1+ cells after ICH (n = 12 slices from 6 mice per group, Student's t test). Scale bar: 100 µm. Hemorrhage borders are demarcated by distinct dashed lines. C) Example gating strategy for flow cytometric analysis of brain immune cells. D–F) Fluorescent‐conjugated surface antibodies were employed to distinguish distinct cell populations: D) Ly6G⁺ neutrophils, E) Ly6G − Ly6C⁺ macrophages/monocytes, and F) CD3e⁺ T cells ( n = 6 per group, Student's t test). G) The heatmap demonstrated alterations in chemokine expression across experimental groups. H) Levels of chemokines CXCL16, CXCL10, CCL7, CXCL1, CXCL9, CCL2, CCL5, and CCL25 in the post‐ICH brain at 72 h were quantified via ELISA ( n = 6 per group, Student's t test). I,J) Flow cytometry assessed the impact of anti‐CCL5 antibody on Ly6G⁺ neutrophil infiltration levels in mouse brains at 24 h after ICH ( n = 6 per group, one‐way ANOVA). (K‐L) Quantification analysis of the CitH3 + cell number per mm 2 ( n = 6 per group, Student's t ‐test). Scale bar: 50 µm. M,N) Representative micrographs of IBA1 and CD86 expression, along with quantitative analysis of CD86+ cells in IBA1+ cells following anti‐CCL5 treatment ( n = 12 slices from 6 mice per group, Student's t ‐test). Scale bar: 100 µm. Data are presented as means ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Moreover, the therapeutic potential of the CCL5‐CCR5 axis was evaluated by intraperitoneal administration of a CCL5‐neutralizing antibody (50 μg/mouse, R&D Systems, MAB478‐500) or a CCR5 antagonist (100 mg kg −1 , Selleckchem, S2003).

Techniques: Immunofluorescence, Staining, Expressing, Enzyme-linked Immunosorbent Assay, Flow Cytometry

Journal: Journal of Nanobiotechnology

Article Title: Apigenin regulates CCR5/JAK1/STAT1/MMPs signaling to alleviate secondary brain injury after intracerebral hemorrhage and its enhanced delivery via targeted nanoparticles

doi: 10.1186/s12951-025-03748-6

Figure Lengend Snippet: Activation of the CCR5-JAK1-STAT1-MMPs Signaling Pathway in the Perihematomal Region Following ICH.( A ) Volcano plot of DEGs. Red indicates upregulated genes, gray indicates non-significant genes, and blue indicates downregulated genes. Screening criteria: p < 0.05 and log₂FC ≥ 0.5; ( B ) Heatmap of DEGs, with red indicating high expression and blue indicating low expression; ( C ) Bar chart of CCL4, CCL5, CCR5, JAK1, STAT3, and MMP3 expression; ( D ) Heatmap of module-trait relationships with ICH and control groups, where red indicates positive correlation and blue indicates negative correlation; ( E - F ) GO and KEGG enrichment analyses of DEGs in the brown module; ( G - H ) GO and KEGG enrichment analyses of DEGs in the yellow module; ( I ) qPCR validation of CCL3, CCL4, and CCL5 expression after ICH; ( J ) WB and quantitative analysis of CCR5, IBA-1, p-JAK1, JAK1, and MMP9 protein levels in brain tissues on days 1, 3, and 7 post-ICH; ( K ) WB analysis of JAK1-STAT1-MMP pathway-related proteins on day 3 post-ICH; ( L ) Evaluation of CCR5 expression in macrophages under different treatment conditions following administration of the CCR5 inhibitor MVC (100 mg/kg). rCCL4 treatment upregulated CCR5 expression, while API showed no significant effect; ( M ) Under rCCL4 stimulation (1 μg/mL), assessment of p-JAK1 expression revealed that MVC (5 μM) inhibited its expression; ( N ) Immunofluorescence analysis of p-JAK1 phosphorylation levels in macrophages following MVC intervention. All data are presented as mean ± SD. Statistical significance was determined using two-way ANOVA followed by Tukey's multiple comparison test. * p < 0.05,** p < 0.01, *** p < 0.001, **** p < 0.0001 vs. ICH group or ICH + 3 d group; ns: not significant

Article Snippet: Maraviroc (MVC), a selective CCR5 antagonist (100 mg/kg for animal experiments or 5 μL for cell experiments, administered for 6 h; S2003, Selleck Chemicals, USA), was dissolved in 1% DMSO.

Techniques: Activation Assay, Expressing, Control, Biomarker Discovery, Immunofluorescence, Phospho-proteomics, Comparison

Journal: Journal of Nanobiotechnology

Article Title: Apigenin regulates CCR5/JAK1/STAT1/MMPs signaling to alleviate secondary brain injury after intracerebral hemorrhage and its enhanced delivery via targeted nanoparticles

doi: 10.1186/s12951-025-03748-6

Figure Lengend Snippet: RVG/FA-NPs@API Alleviates Neurotoxicity by Inhibiting the CCR5/JAK1/STAT1/MMPs Pathway in Macrophages.( A ) Schematic diagram illustrating the LPS-induced macrophage inflammation model, the intervention of RVG/FA-NPs@API, and its neuroprotective mechanism, created in BioRender; ( B ) mRNA expression level of CCR5 in RAW264.7 cells detected by RT-qPCR; ( C ) Protein expression levels of CCR5, JAK1, p-JAK1, STAT1, p-STAT1, MMP9, and MMP3 in macrophages determined by WB; ( D ) Protein expression levels of TNF-α and IL-1β in RAW264.7 cells detected by WB; ( E ) Immunofluorescence staining of iNOS and Arg1 in RAW264.7 cells, scale bar = 25 μm; ( F ) Cell viability of HT-22 neurons after co-culture with different treatment groups assessed using the CCK-8 assay; ( G ) Neuronal apoptosis and death assessed using Calcein-AM/PI double staining, scale bar = 50 μm; ( H ) Expression levels of Bax, Bcl-2, NF200, PSD95, and GAP43 proteins in neurons detected by WB. All cell experiments were repeated three times. * p < 0.05, ** p < 0.01, *** p < 0.001,**** p < 0.0001 between groups

Article Snippet: Maraviroc (MVC), a selective CCR5 antagonist (100 mg/kg for animal experiments or 5 μL for cell experiments, administered for 6 h; S2003, Selleck Chemicals, USA), was dissolved in 1% DMSO.

Techniques: Expressing, Quantitative RT-PCR, Immunofluorescence, Staining, Co-Culture Assay, CCK-8 Assay, Double Staining

Journal: Journal of Nanobiotechnology

Article Title: Apigenin regulates CCR5/JAK1/STAT1/MMPs signaling to alleviate secondary brain injury after intracerebral hemorrhage and its enhanced delivery via targeted nanoparticles

doi: 10.1186/s12951-025-03748-6

Figure Lengend Snippet: RVG/FA-NPs@API Alleviates SBI in ICH Mice. ( A ) Schematic diagram of the experimental workflow illustrating the intervention strategy and group design of RVG/FA-NPs@API in the ICH model, created in BioRender; ( B ) BWC in each group measured by the wet-dry weight method; ( C ) Pathological changes in brain tissue assessed by H&E staining, scale bar = 100 μm; ( D ) Neuronal arrangement and density in brain tissue evaluated by Nissl staining, scale bar = 100 μm; ( E ) Proportion of apoptotic neurons in brain tissue detected by TUNEL staining, with TUNEL-positive cells shown in green, scale bar = 50 μm; ( F ) Expression levels of iNOS (green) and Arg1 (red) in brain tissue detected by immunofluorescence, scale bar = 25 μm; ( G ) Protein expression levels of CCR5, p-JAK1, p-STAT1, MMP9, MMP3, ZO-1, and CLDN5 in brain tissue detected by WB. Each group included n = 6 animals. * p < 0.05, ** p < 0.01, *** p < 0.001 between groups

Article Snippet: Maraviroc (MVC), a selective CCR5 antagonist (100 mg/kg for animal experiments or 5 μL for cell experiments, administered for 6 h; S2003, Selleck Chemicals, USA), was dissolved in 1% DMSO.

Techniques: Staining, TUNEL Assay, Expressing, Immunofluorescence