ccr2 inhibitor rs504393  (MedChemExpress)

Journal: The Journal of Clinical Investigation

Article Title: Inhibition of aortic CX3CR1 + macrophages mitigates thoracic aortic aneurysm progression in Marfan syndrome in mice

doi: 10.1172/JCI178198

Figure Lengend Snippet: ( A – D ) Six-week-old Fbn1 C1041G/+ mice were intraperitoneally injected with RS504393 (4 mg/kg/d) until age 26 weeks. Age-matched WT mice were used as a normal control. ( A ) Flow cytometry analysis of Ly6C + monocytes and CX3CR1 + macrophages in aortic root and ascending aortas. n = 4. * P < 0.05 by 1-way ANOVA followed by Tukey’s test for post hoc comparison. ( B ) Representative transthoracic echocardiographic images of the aortic root and ascending aortas. Scale bars: 2 mm. White arrows depict the sinus of Valsalva measurements. Yellow arrows depict ascending aorta measurement. ( C ) Quantification of transthoracic echocardiography. n = 4 for WT and n = 8 for Fbn1 C1041G/+ mice. * P < 0.05 by repeated-measures ANOVA with the Greenhouse-Geisser adjustment followed by Bonferroni’s post hoc comparisons. ( D ) EVG staining of the aortic roots. n = 5. * P < 0.05 by Kruskal-Wallis test followed by Dunn’s test for elastin degradation grade and 1-way ANOVA followed by Tukey’s test for post hoc comparison for aortic thickness. ( E – H ) Twenty-four-week-old Fbn1 C1041G/+ mice were intraperitoneally injected with RS504393 (4 mg/kg/d) until age 36 weeks. Age-matched WT mice were used as a normal control. ( E ) Flow cytometry analysis of Ly6C + monocytes and CX3CR1 + macrophages in aortic root and ascending aortas. n = 5. * P < 0.05 by 1-way ANOVA followed by Tukey’s test for post hoc comparison. ( F ) Representative transthoracic echocardiographic images of the aortic root and ascending aortas. Scale bars: 2 mm. White arrows depict the sinus of Valsalva measurements. Yellow arrows depict ascending aorta measurement. ( G ) Quantification of transthoracic echocardiography. n = 5. * P < 0.05 by repeated-measures ANOVA with the Greenhouse-Geisser adjustment followed by Bonferroni’s post hoc comparisons. ( H ) EVG staining of aortic roots. n = 5. * P < 0.05 by Kruskal-Wallis test followed by Dunn’s test for elastin degradation grade and 1-way ANOVA followed by Tukey’s test for post hoc comparison for aortic thickness.

Article Snippet: Tamoxifen (HY-13757A), adalimumab (HY-P9908), teprotumumab (HY-P99165), and the CCR2 inhibitor RS504393 (HY-15418) were purchased from MedChem Express.

Techniques: Injection, Control, Flow Cytometry, Comparison, Staining

Journal: The Journal of Clinical Investigation

Article Title: Inhibition of aortic CX3CR1 + macrophages mitigates thoracic aortic aneurysm progression in Marfan syndrome in mice

doi: 10.1172/JCI178198

Figure Lengend Snippet: CX3CR1 + macrophages mainly located in the aortic intima and originated from circulating Ly6C + monocytes. Intimal CX3CR1 + macrophages upregulated inflammatory genes, including IL6 , MCP1 , and CXCL2 , in VSMCs by producing and secreting TNF-α and IGF1, thereby aggravating TAA development in MFS. Either elimination of intimal CX3CR1 + macrophages or administration of a CCR2 inhibitor to suppress monocyte recruitment efficiently alleviated TAA progression in MFS.

Article Snippet: Tamoxifen (HY-13757A), adalimumab (HY-P9908), teprotumumab (HY-P99165), and the CCR2 inhibitor RS504393 (HY-15418) were purchased from MedChem Express.

Techniques:

Journal: Neuroreport

Article Title: Hyperbaric oxygen therapy improves neurological function via the p38-MAPK/CCL2 signaling pathway following traumatic brain injury

doi: 10.1097/WNR.0000000000001719

Figure Lengend Snippet: Primer sequences

Article Snippet: The CCR2 antagonist (RS504393) (Tocris, Bristol, South West England, UK) or p38 inhibitor (SB203580) (Calbiochem, Merck, Darmstadt, Germany) were dissolved in dimethyl sulfoxide (DMSO) and diluted with PBS to a low dose (2.5 μg/10 μL) and a high dose (25 μg/10 μL) and then injected into the injured area, respectively, at 1 h following TBI as described previously [ ].

Techniques:

Journal: Neuroreport

Article Title: Hyperbaric oxygen therapy improves neurological function via the p38-MAPK/CCL2 signaling pathway following traumatic brain injury

doi: 10.1097/WNR.0000000000001719

Figure Lengend Snippet: TBI-induced upregulation of CCL2, CCR2, and p-p38 expression in the injured cortex of TBI rats. (a) Expression of CCL2 protein peaks on the third day after TBI, compared to sham group ( n = 5/group). (b). Expression of CCR2 protein peaks on the third day after TBI, compared to sham group ( n = 5/group). (c) The expression of p-p38 decreased after peaking on the first day after TBI ( n = 3/group). Values are expressed as mean ± SEM. *** P < 0.001, ** P < 0.01, * P < 0.05 vs. sham group. TBI, traumatic brain injury.

Article Snippet: The CCR2 antagonist (RS504393) (Tocris, Bristol, South West England, UK) or p38 inhibitor (SB203580) (Calbiochem, Merck, Darmstadt, Germany) were dissolved in dimethyl sulfoxide (DMSO) and diluted with PBS to a low dose (2.5 μg/10 μL) and a high dose (25 μg/10 μL) and then injected into the injured area, respectively, at 1 h following TBI as described previously [ ].

Techniques: Expressing

Journal: Neuroreport

Article Title: Hyperbaric oxygen therapy improves neurological function via the p38-MAPK/CCL2 signaling pathway following traumatic brain injury

doi: 10.1097/WNR.0000000000001719

Figure Lengend Snippet: The neurological and cognitive function of TBI rats was improved by CCR2 antagonist (RS504393) or p38 inhibitor (SB203580). (a) The mNSS score of TBI rats decreased after the application of high-dose CCR2 antagonists. (b) The mNSS score of TBI rats decreased after the application of high-dose p38 inhibitor. (c) The escape latency of TBI rats was reduced after the application of a high-dose CCR2 antagonist or p38 inhibitor. (d) The number of platform crossings of TBI rats increased after the application of high-dose CCR2 antagonist or p38 inhibitor. Values are expressed as mean ± SEM ( n = 8/group). *** P < 0.001, * P < 0.05 vs. TBI + vehicle group. ### P < 0.001, ## P < 0.01, vs. sham group. TBI, traumatic brain injury; mNSS, modified neurological severity score.

Article Snippet: The CCR2 antagonist (RS504393) (Tocris, Bristol, South West England, UK) or p38 inhibitor (SB203580) (Calbiochem, Merck, Darmstadt, Germany) were dissolved in dimethyl sulfoxide (DMSO) and diluted with PBS to a low dose (2.5 μg/10 μL) and a high dose (25 μg/10 μL) and then injected into the injured area, respectively, at 1 h following TBI as described previously [ ].

Techniques: Modification

Journal: Neuroreport

Article Title: Hyperbaric oxygen therapy improves neurological function via the p38-MAPK/CCL2 signaling pathway following traumatic brain injury

doi: 10.1097/WNR.0000000000001719

Figure Lengend Snippet: p38 inhibitor (SB203580) downregulated CCL2 and CCR2 mRNA expression. CCL2 and CCR2 mRNA expression decreased significantly at 3 days after continuous injection of high doses of p38 inhibitor. Values are expressed as mean ± SEM ( n = 6/group). ** P < 0. 01, * P < 0.05, vs. TBI + vehicle group; ### P < 0.001, vs. sham group. TBI, traumatic brain injury.

Article Snippet: The CCR2 antagonist (RS504393) (Tocris, Bristol, South West England, UK) or p38 inhibitor (SB203580) (Calbiochem, Merck, Darmstadt, Germany) were dissolved in dimethyl sulfoxide (DMSO) and diluted with PBS to a low dose (2.5 μg/10 μL) and a high dose (25 μg/10 μL) and then injected into the injured area, respectively, at 1 h following TBI as described previously [ ].

Techniques: Expressing, Injection

Journal: Neuroreport

Article Title: Hyperbaric oxygen therapy improves neurological function via the p38-MAPK/CCL2 signaling pathway following traumatic brain injury

doi: 10.1097/WNR.0000000000001719

Figure Lengend Snippet: CCL2 and CCR2 mRNA expression and p-p38 expression were downregulated after HBO treatment. (a) mRNA expression of CCL2 was downregulated after 1, 3, 7 days of HBO treatment. CCL2 mRNA expression was downregulated after 10 days of HBO treatment, but the difference was not statistically significant ( n = 6/group). (b) mRNA expression of CCR2 was downregulated after 1, 3, 7 days of HBO treatment. CCR2 mRNA expression was downregulated after 10 days of HBO treatment, but the difference was not statistically significant ( n = 6/group). (c) The expression of p-p38 significantly decreased after continuous HBO treatment for 3 days ( n = 3/group). Values are expressed as mean ± SEM. *** P < 0.001, ** P < 0.01, * P < 0.05 vs. TBI group. HBO, hyperbaric oxygen; TBI, traumatic brain injury.

Article Snippet: The CCR2 antagonist (RS504393) (Tocris, Bristol, South West England, UK) or p38 inhibitor (SB203580) (Calbiochem, Merck, Darmstadt, Germany) were dissolved in dimethyl sulfoxide (DMSO) and diluted with PBS to a low dose (2.5 μg/10 μL) and a high dose (25 μg/10 μL) and then injected into the injured area, respectively, at 1 h following TBI as described previously [ ].

Techniques: Expressing

Journal: Advanced Science

Article Title: CCN1 Enhances Tumor Immunosuppression through Collagen‐Mediated Chemokine Secretion in Pancreatic Cancer

doi: 10.1002/advs.202500589

Figure Lengend Snippet: Collagens modulate chemokine expression in PDAC cells under IFNγ stimulation. A) Tumor growth of sgCtrl and sgCcn1 KPC cells inoculated subcutaneously in C57B/L6 mice. Tumor‐bearing mice were treated with CCR2 inhibitor RS504393 and CXCR2 inhibitor SB225002. n = 6 mice per group. B) Representative images of tumors (left) and quantification of tumor weight (right) in mice inoculated with sgCtrl or sgCcn1 KPC cells. Tumor‐bearing mice were treated with the CCR2 inhibitor RS504393 and the CXCR2 inhibitor SB225002. C) Flow cytometric analysis of MDSC proportions in subcutaneous sgCtrl and sgCcn1 KPC tumors. D) mRNA levels of collagens, including Col4a1, Cola4a2, Col5a1, Col6a1, Col6a2, Col6a3, and Col8a1, in KPC cells after their total knockdown. E) mRNA levels of chemokines, including Ccl2, Ccl20, Cxcl1, Cxcl3, Cxcl5, Csf1, Csf2, and Csf 3 , in KPC cells after their total knockdown. F–H) mRNA levels of chemokines, including Ccl2, Ccl7, Ccl20, Cxcl1, Cxcl3, Cxcl5, Csf1, Csf2, and Csf3, in KPC cells after Col5a1 knockdown (F), Col6a3 knockdown (G), and Col8a1 knockdown (H). I) mRNA levels of collagens, including Col4a1, Cola4a2, Col5a1, Col6a1, Col6a2, Col6a3, and Col8a1, in sgCtrl KPC cells after treatment with 100 ng mL −1 IFNγ. J) mRNA levels of Col5a1, Col6a3, and Col8a1 measured in siCol5a1, siCol6a3, and siCol8a1 KPC cells after treatment with 100 ng mL −1 IFNγ. K–M) mRNA levels of chemokines including the Ccls (K), Cxcls (L), and Csfs (M), in siCol5a1, siCol6a3, and siCol8a1 KPC cells after treatment with 100 ng mL −1 IFNγ. N) mRNA level of Il6 in siCol5a1, siCol6a3, and siCol8a1 KPC cells after treatment with 100 ng mL −1 IFNγ.

Article Snippet: CCR2 inhibitor RS504393 (S9738, Selleck), CXCR2 inhibitor SB225002 (S7651, Selleck), and gemcitabine (S1714, Selleck) were purchased from Selleckchem.

Techniques: Expressing, Knockdown

Journal: Advanced Science

Article Title: CCN1 Enhances Tumor Immunosuppression through Collagen‐Mediated Chemokine Secretion in Pancreatic Cancer

doi: 10.1002/advs.202500589

Figure Lengend Snippet: Ccn1 regulates the TME in PDAC. A) Schematic representation of macrophage isolation from mouse bone marrow. B) Macrophage invasion evaluated by the chemotactic effect of sgCtrl and sgCcn1 KPC cells in a Transwell invasion assay. C) Macrophage polarization assessed by flow cytometry in macrophages cocultured with sgCtrl and sgCcn1 KPC cells. D) Macrophage polarization assessed by flow cytometry in macrophages cocultured with sgCtrl and sgCcn1 KPC cells after treatment with CCR2 inhibitor RS504393 and CXCR2 inhibitor SB225002. E) Survival of sgCtrl and sgCcn1 KPC cells cocultured with macrophages, assessed by CCK‐8 assay. F) Cell death of sgCtrl and sgCcn1 KPC cells cocultured with macrophages, assessed by flow cytometry. G) Protein expression levels of TNFα in macrophages cocultured with sgCtrl and sgCcn1 KPC cells. H) mRNA expression levels of TNFα in macrophages cocultured with sgCtrl and sgCcn1 KPC cells. I) Protein expression level of c‐Caspase 3 in sgCtrl and sgCcn1 KPC cells after treatment with 100 ng mL −1 TNFα. J) Protein expression level of c‐Caspase 3 in sgCtrl and sgCcn1 KPC cells after treatment with 100 ng mL −1 TNFα, mouse CCN1 protein, and cilengitide (αVβ3 and αVβ5 integrin inhibitor). K) Nucleus localization of CCN1 in KPC cells with CCN1 (red) and DAPI (blue) fluorescence. L) Protein expression level of p‐STAT3 in sgCtrl and sgCcn1 KPC cells after treatment with 100 ng mL −1 INFγ. M) mRNA levels of Ccl7, Cxcl3, and Csf3 in sgCtrl and sgCcn1 KPC cells after treatment with 100 ng mL −1 INFγ. N) mRNA level of Il6 measured in sgCtrl and sgCcn1 KPC cells after treatment with 100 ng mL −1 INFγ.

Article Snippet: CCR2 inhibitor RS504393 (S9738, Selleck), CXCR2 inhibitor SB225002 (S7651, Selleck), and gemcitabine (S1714, Selleck) were purchased from Selleckchem.

Techniques: Isolation, Transwell Invasion Assay, Flow Cytometry, CCK-8 Assay, Expressing, Fluorescence

Journal: The Journal of Pharmacology and Experimental Therapeutics

Article Title: Transport of the Proinflammatory Chemokines C-C Motif Chemokine Ligand 2 (MCP-1) and C-C Motif Chemokine Ligand 5 (RANTES) across the Intact Mouse Blood-Brain Barrier Is Inhibited by Heparin and Eprodisate and Increased with Systemic Inflammation

doi: 10.1124/jpet.122.001380

Figure Lengend Snippet: Effects of CCR2 and CCR5 inhibitors on the brain uptake of 125I-CCL2 and 125I-CCL5 in situ. Mice were coperfused with a 125I-chemokine and 99mTc-Alb with 10 uM of the corresponding inhibitor (RS504393 for CCL2 and maraviroc for CCL5, open triangles) or DMSO vehicle (open circles). Each point was corrected for luminal vascular space by subtracting the 99mTc-Alb brain/perfusate ratios. The corresponding data and statistical analysis for this figure are presented in Table 4.

Article Snippet: In some experiments, freshly prepared CCR2 inhibitor RS504393 (SML0711) (Sigma-Aldrich) or CCR5 inhibitor maraviroc (PZ0002) (Sigma-Aldrich) or vehicle (dimethylsulfoxide) were included.

Techniques: In Situ

Journal: The Journal of Pharmacology and Experimental Therapeutics

Article Title: Transport of the Proinflammatory Chemokines C-C Motif Chemokine Ligand 2 (MCP-1) and C-C Motif Chemokine Ligand 5 (RANTES) across the Intact Mouse Blood-Brain Barrier Is Inhibited by Heparin and Eprodisate and Increased with Systemic Inflammation

doi: 10.1124/jpet.122.001380

Figure Lengend Snippet: Effects of CCR2 (RS504393) and CCR5 (maraviroc) inhibitors on 125 I-CCL2 and 125 I-CCL5 brain uptake in situ

Article Snippet: In some experiments, freshly prepared CCR2 inhibitor RS504393 (SML0711) (Sigma-Aldrich) or CCR5 inhibitor maraviroc (PZ0002) (Sigma-Aldrich) or vehicle (dimethylsulfoxide) were included.

Techniques:

Journal: Frontiers in Immunology

Article Title: Human IL-17 and TNF-α Additively or Synergistically Regulate the Expression of Proinflammatory Genes, Coagulation-Related Genes, and Tight Junction Genes in Porcine Aortic Endothelial Cells

doi: 10.3389/fimmu.2022.857311

Figure Lengend Snippet: The combination of IL-17 with TNF-α largely increased the chemotaxis of THP-1 cells and human neutrophils. (A) Porcine aortic endothelial cells (PAECs) were treated with rhIL-17 (100 ng/ml), rhTNF-α (2 ng/ml), or rhIL-17 plus rhTNF-α for 0 or 6 h. The induction of CXCL8 or CCL2 mRNA was measured via real-time PCR. (B) The PAECs were treated with IL-17, TNF-α, or IL-17 plus TNF-α for 48 h, and the supernatant was collected for chemotaxis assays. Human neutrophils or THP‐1 cells were used to assess cell migration. (C) The number of migrating cells per field was determined as in (B) . Data are representative of at least three independent experiments (mean ± SEM). * p < 0.05, ** p < 0.01, *** p < 0.001, determined by Student’s t -test.

Article Snippet: The CCR2 (CCL2 receptor)-specific inhibitor RS504393 was from MedChemExpress (Shanghai, China).

Techniques: Chemotaxis Assay, Real-time Polymerase Chain Reaction, Migration

Journal: Frontiers in Immunology

Article Title: Human IL-17 and TNF-α Additively or Synergistically Regulate the Expression of Proinflammatory Genes, Coagulation-Related Genes, and Tight Junction Genes in Porcine Aortic Endothelial Cells

doi: 10.3389/fimmu.2022.857311

Figure Lengend Snippet: IL-17 and IL-1β or IFN-γ additively or synergistically induced the expression of certain proinflammatory genes in porcine aortic endothelial cells (PAECs). (A) The PAECs were treated with rhIL-17 (50 ng/ml), rhIL-1β (10 ng/ml), or rhIL-17 plus rhIL-1β for 0, 2, or 6 h. The induction of E-selectin, VCAM-1, ICAM-1, IL-6, IL-8, MCP-1, or CXCL2 mRNA was measured using real-time PCR. (B) The PAECs were treated with rhIL-17 (50 ng/ml), rpIFN-γ (40 ng/ml), or rhIL-17 plus rpIFN-γ for 0, 2, or 6 h. The induction of E-selectin, VCAM-1, ICAM-1, IL-6, IL-8, MCP-1, or CXCL2 mRNA was measured using real-time PCR. Data are representative of at least three independent experiments (mean ± SEM). * p < 0.05, ** p < 0.01, *** p < 0.001, determined by Student’s t -test.

Article Snippet: The CCR2 (CCL2 receptor)-specific inhibitor RS504393 was from MedChemExpress (Shanghai, China).

Techniques: Expressing, Real-time Polymerase Chain Reaction

Journal: Communications Biology

Article Title: Inhibition of the CCL2 receptor, CCR2, enhances tumor response to immune checkpoint therapy

doi: 10.1038/s42003-020-01441-y

Figure Lengend Snippet: a Subcutaneous tumor growth in syngeneic mice injected with NA13 cells ( n = 6 biologically independent mice per group). Data representative of two independent experiments. Statistical significance was determined by two-way ANOVA. Arrows indicate dates in which anti-PD-1 treatment was given. Mean ± SD. b Individual tumor volumes as a function of time. Each line represents a single mouse ( n = 10 or 11 biologically independent mice per group). c Waterfall plot showing change in NA13 tumor volume on endpoint day 30 post tumor injection compared to baseline prior to treatment with and without CCR2 antagonist RS504393 and/or anti-PD-1 treatment. Starting on day 14, mice were treated daily with CCR2 antagonist RS503393, and every three days with anti-PD-1.

Article Snippet: CCR2 small molecule inhibitor RS504393 (Tocris) was given daily at 2 mg/kg by oral gavage to B16F10 mice and intraperitoneally to E0771 and NA13 mice.

Techniques: Injection