Journal: Journal of Enzyme Inhibition and Medicinal Chemistry

Article Title: Discovery of 2-phenylethyl chromones as potent and selective CYP1B1 inhibitors

doi: 10.1080/14756366.2025.2598738

Figure Lengend Snippet: Design of 2–(2-phenylethyl)-chromone derivatives for CYP1B1 inhibitors.

Article Snippet: Mouse monoclonal antibody to CYP1B1 (Proteintech, Cat. No. 67033-1-Ig) was used at a dilution of 1:6000, while mouse monoclonal antibody to β-actin (Proteintech, Cat. No. 66009-1-Ig) was applied at 1:10000 dilution.

Techniques:

Journal: Journal of Enzyme Inhibition and Medicinal Chemistry

Article Title: Discovery of 2-phenylethyl chromones as potent and selective CYP1B1 inhibitors

doi: 10.1080/14756366.2025.2598738

Figure Lengend Snippet: Structure–activity relationship (SAR) of 2–(2-phenylethyl) chromone derivatives against CYP1B1 activity.

Article Snippet: Mouse monoclonal antibody to CYP1B1 (Proteintech, Cat. No. 67033-1-Ig) was used at a dilution of 1:6000, while mouse monoclonal antibody to β-actin (Proteintech, Cat. No. 66009-1-Ig) was applied at 1:10000 dilution.

Techniques: Activity Assay

Journal: Journal of Enzyme Inhibition and Medicinal Chemistry

Article Title: Discovery of 2-phenylethyl chromones as potent and selective CYP1B1 inhibitors

doi: 10.1080/14756366.2025.2598738

Figure Lengend Snippet: Construction of MCF-7/ cyp1b1 cell lines via MCF-7 infected lentivirus carrying cyp1b1 and green fluorescence. (MCF-7 cells and MCF-7/ cyp1b1 cells were all nuclear stained by DAPI for 15 min and washed by PBS for three times. Then, they were observed and photographed under the fluorescence microscope. Blue fluorescence indicates DAPI nuclear staining, Green fluorescence indicates CYP1B1 mutant expressing.).

Article Snippet: Mouse monoclonal antibody to CYP1B1 (Proteintech, Cat. No. 67033-1-Ig) was used at a dilution of 1:6000, while mouse monoclonal antibody to β-actin (Proteintech, Cat. No. 66009-1-Ig) was applied at 1:10000 dilution.

Techniques: Infection, Fluorescence, Staining, Microscopy, Mutagenesis, Expressing

Journal: Journal of Enzyme Inhibition and Medicinal Chemistry

Article Title: Discovery of 2-phenylethyl chromones as potent and selective CYP1B1 inhibitors

doi: 10.1080/14756366.2025.2598738

Figure Lengend Snippet: Expression of CYP1B1 enzyme detected by western-blotting in MCF-7, MCF-7/DMBA and MCF-7/ cyp1b1 cell lines; Survival curve of cells treated with docetaxel.

Article Snippet: Mouse monoclonal antibody to CYP1B1 (Proteintech, Cat. No. 67033-1-Ig) was used at a dilution of 1:6000, while mouse monoclonal antibody to β-actin (Proteintech, Cat. No. 66009-1-Ig) was applied at 1:10000 dilution.

Techniques: Expressing, Western Blot

Journal: Journal of Enzyme Inhibition and Medicinal Chemistry

Article Title: Discovery of 2-phenylethyl chromones as potent and selective CYP1B1 inhibitors

doi: 10.1080/14756366.2025.2598738

Figure Lengend Snippet: Reversal of drug resistance by chromones in CYP1B1-overexpressing cell lines. (A. Three chromones reversed drug resistance of MCF-7/c yp1b1 and MCF-7/DMBA cell lines to docetaxel (DOC); B. CX-9 reversed MCF-7/ cyp1b1 resistant to DOC dose-dependently.). The data were expressed as the mean ± SD ( n = 3). * p < 0.05 compared with the docetaxel-treating cells.

Article Snippet: Mouse monoclonal antibody to CYP1B1 (Proteintech, Cat. No. 67033-1-Ig) was used at a dilution of 1:6000, while mouse monoclonal antibody to β-actin (Proteintech, Cat. No. 66009-1-Ig) was applied at 1:10000 dilution.

Techniques:

Journal: Journal of Enzyme Inhibition and Medicinal Chemistry

Article Title: Discovery of 2-phenylethyl chromones as potent and selective CYP1B1 inhibitors

doi: 10.1080/14756366.2025.2598738

Figure Lengend Snippet: Molecule docking and molecular dynamics simulation of CYP1B1 (PDB ID: 3PM0) and CX-9 and ANF, respectively. (A. Docking pose of CX-9 (green sticks) in the CYP1B1-ANF complex. Phe 231 (purple), the haem prosthetic group (violet), and Gly 329 (yellow) are shown. B. 2D docked model of CX-9 into CYP1B1, hydrogen bonds of Asp 333 , Phe 231 and Gly 329 are shown by dashed lines. C. Superimposed structures of CYP1B1 enzyme bound to CX-9 and ANF. D. Protein RMSD, backbone RMSD (fit to protein) over 100 ns for apo (cyan), ANF (purple), and CX-9 (green). E. RMSF by residue. RMSF (Å) highlighting B-C (∼110–130), I-helix/SRS-4 (∼290–370), F-G (∼370–410), and haem-binding (∼430-470) regions. F. Protein-ligand hydrogen bonds. Instantaneous hydrogen-bond counts using a 0.35 nm/135° geometric criterion.).

Article Snippet: Mouse monoclonal antibody to CYP1B1 (Proteintech, Cat. No. 67033-1-Ig) was used at a dilution of 1:6000, while mouse monoclonal antibody to β-actin (Proteintech, Cat. No. 66009-1-Ig) was applied at 1:10000 dilution.

Techniques: Residue, Binding Assay

Journal: Molecular Medicine Reports

Article Title: Exploring the role of cytochrome P450 family 1 subfamily B member 1 and quercetin in modulating neuropathic pain after spinal cord injury

doi: 10.3892/mmr.2025.13717

Figure Lengend Snippet: Research workflow. CYP1B1, cytochrome P450 family 1 subfamily B member 1; eQTL, expression-quantitative trait loci; HEIDI, heterogeneity in dependent instruments; NP, neuropathic pain; PPI, protein-protein interaction; Que, quercetin; SCI, spinal cord injury; SMR, summary-based Mendelian randomization; WGCNA, weighted gene co-expression network analysis.

Article Snippet: The protein samples were then transferred to polyvinylidene fluoride membranes, which were subsequently blocked for 1 h at room temperature., washed with TBST solution and incubated with the following primary antibodies: CYP1B1 (1:500; cat. no. 18505-1-AP; Proteintech Group, Inc.), Fibronectin (Fn) (1:1,000; cat. no. ab2413; Abcam) and GAPDH (1:50,000; cat. no. 1E6D9; Proteintech Group, Inc.).

Techniques: Expressing

Journal: Molecular Medicine Reports

Article Title: Exploring the role of cytochrome P450 family 1 subfamily B member 1 and quercetin in modulating neuropathic pain after spinal cord injury

doi: 10.3892/mmr.2025.13717

Figure Lengend Snippet: (A) Screening of diagnostic marker genes in the LASSO model. This panel shows the cross-validated mean-squared error plotted against Log(λ). The number of genes corresponding to the lowest point of the curve (n=10) was most suitable for the diagnosis of SCI. (B) This panel displays the number of features (variables) retained in the model at different values of Log(λ). (C) Characteristic gene selection using the SVM-RFE technique. A total of 11 characteristic genes were identified from the 21 core genes. (D) The 15 trait genes were ranked according to the importance score. (E) Venn diagram of diagnostic candidate genes identified by three machine algorithms. (F) Predicted nomograms of candidate genes for SCI diagnosis. To use it, locate the value for each biomarker (GPD1L, EPHX2, CYP1B1) on its respective scale and draw a line upward to the ‘Points’ axis to determine the score for each variable. Sum all the points to get the ‘Total points’. (G) ROC curve of CYP1B1 in GSE151371 with an AUC value of 0.953. (H) ROC curve of EPHX2 in GSE151371 with an AUC value of 0.989. (I) ROC curve of GPD1L in GSE151371 with an AUC value of 0.957. (J) Visualization of SCI diagnostic candidate genes based on CTD pain inference scores. AUC, area under the curve; CTD, Comparative Toxicogenomics Database; CYP1B1, cytochrome P450 family 1 subfamily B member 1; EPHX2, epoxide hydrolase 2; GPD1L, glycerol-3-phosphate dehydrogenase 1-like; LASSO, least absolute shrinkage and selection operator; RMSE, root mean square error; ROC, receiver operating characteristic; SCI, spinal cord injury; SVM-RFE, support vector machine recursive feature elimination.

Article Snippet: The protein samples were then transferred to polyvinylidene fluoride membranes, which were subsequently blocked for 1 h at room temperature., washed with TBST solution and incubated with the following primary antibodies: CYP1B1 (1:500; cat. no. 18505-1-AP; Proteintech Group, Inc.), Fibronectin (Fn) (1:1,000; cat. no. ab2413; Abcam) and GAPDH (1:50,000; cat. no. 1E6D9; Proteintech Group, Inc.).

Techniques: Diagnostic Assay, Marker, Biomarker Discovery, Selection, Plasmid Preparation

Journal: Molecular Medicine Reports

Article Title: Exploring the role of cytochrome P450 family 1 subfamily B member 1 and quercetin in modulating neuropathic pain after spinal cord injury

doi: 10.3892/mmr.2025.13717

Figure Lengend Snippet: (A) MR association between CYP1B1 gene expression and trigeminal neuralgia in blood. All SNPs available in GWAS and eQTL data are shown in. (A) Grey dots represent the P-values of SNPs for GWAS, and diamonds represent P-values of probes for SMR tests. (B) P-values for eQTL analyses of CYP1B1 SNPs from GWAS (used in the heterogeneity in dependent instruments test) are plotted against effect sizes of SNPs from eQTL studies. The orange dashed line indicates the effect size estimate for the MR association at the top cis-eQTL. The error line is the standard error of the SNP effect. (C) Chemical structure of Que. (D) Interaction of the CYP1B1 receptor with Que ligands. The protein is depicted as a blue cartoon (α-helices and β-sheets) with its molecular surface shown in transparent gray. Que is shown as red sticks. (E) Detailed view of the local interactions within the Que-CYP1B1 binding pocket Protein residues are shown as blue sticks. The ligand, Que, is shown as red sticks. (F) Venn diagram of potential targets of Que and 21 core genes. (G) Rg of the Que-CYP1B1 complex. (H) RMSD of the Que-CYP1B1 complex. (I) RMSF of CYP1B1 protein residues. (J) Solvent accessible surface of the Que-CYP1B1 complex. (K) Free energy landscape of the Que-CYP1B1 complex. CYP1B1, cytochrome P450 family 1 subfamily B member 1; eQTL, expression-quantitative trait loci; GWAS, genome wide association study; MR, Mendelian randomization; PC, principal component; Que, quercetin; Rg, radius of gyration; RMSD, root mean square deviation; RMSF, root mean square fluctuation; SMR, summary-based Mendelian randomization.

Article Snippet: The protein samples were then transferred to polyvinylidene fluoride membranes, which were subsequently blocked for 1 h at room temperature., washed with TBST solution and incubated with the following primary antibodies: CYP1B1 (1:500; cat. no. 18505-1-AP; Proteintech Group, Inc.), Fibronectin (Fn) (1:1,000; cat. no. ab2413; Abcam) and GAPDH (1:50,000; cat. no. 1E6D9; Proteintech Group, Inc.).

Techniques: Gene Expression, Binding Assay, Solvent, Expressing, GWAS

Journal: Molecular Medicine Reports

Article Title: Exploring the role of cytochrome P450 family 1 subfamily B member 1 and quercetin in modulating neuropathic pain after spinal cord injury

doi: 10.3892/mmr.2025.13717

Figure Lengend Snippet: (A) A clustering heat map of the samples with the optimal clustering number set to k=2. (B) CDF curve for various values of k, constrained within the minimum range of the consensus index. (C) Principal component analysis graphs with the optimal clustering number set to k=2. (D) Differential expression of CYP1B1 across two distinct clusters. (E) CYP1B1 expression in the GSE171441 dataset. (F) CYP1B1 expression in the GSE236754 dataset. (G) A total of 13 cell populations were identified in the single-cell sequencing data from the GSE162610 dataset. (H) Proportion of the 13 cell populations in the two groups. (I) CYP1B1 expression in 13 cell populations. (J) Comparison of changes in CYP1B1 levels in fibroblasts of two groups. *P<0.05, **P<0.01, ***P<0.001 and ****P<0.0001. CDF, cumulative distribution function; CYP1B1, cytochrome P450 family 1 subfamily B member 1; NK, natural killer; OPC, oligodendrocyte precursor cells; SCI, spinal cord injury; SHAM, sham surgery group; umap, uniform manifold approximation and projection; Y1, Principal Component 1; Y2, Principal Component 2.

Article Snippet: The protein samples were then transferred to polyvinylidene fluoride membranes, which were subsequently blocked for 1 h at room temperature., washed with TBST solution and incubated with the following primary antibodies: CYP1B1 (1:500; cat. no. 18505-1-AP; Proteintech Group, Inc.), Fibronectin (Fn) (1:1,000; cat. no. ab2413; Abcam) and GAPDH (1:50,000; cat. no. 1E6D9; Proteintech Group, Inc.).

Techniques: Quantitative Proteomics, Expressing, Sequencing, Comparison

Journal: Molecular Medicine Reports

Article Title: Exploring the role of cytochrome P450 family 1 subfamily B member 1 and quercetin in modulating neuropathic pain after spinal cord injury

doi: 10.3892/mmr.2025.13717

Figure Lengend Snippet: (A) Flow chart of the animal experiment. (B) MWT measurements for the SHAM, SCI and SCI + Que groups were recorded on day 14. (C) TWL for the SHAM, SCI and SCI + Que groups was also measured on day 14. (D) BMS scores for the SHAM, SCI and SCI + Que groups were assessed at baseline and on postoperative days 1, 3, 5, 7, 10 and 14. A statistically significant difference was observed (***P<0.001) between the SCI and SHAM groups of mice (n=6 each). Additionally, a significant difference was noted between the SCI and SCI + Que groups (*P<0.05), with n=6. (E) RT-qPCR assessment of CYP1B1 mRNA levels in spinal cord tissues from SHAM, SCI and SCI + Que groups, with expression levels normalized to GAPDH. (F) Semi-quantitative analysis of CYP1B1 protein levels in spinal cord tissues from the SHAM, SCI and SCI + Que groups, with fold-changes normalized to GAPDH. (G) Western blot analysis was conducted to assess CYP1B1 expression in spinal cord tissues from the SHAM, SCI and SCI + Que groups on day 14 post-modeling. (H) Immunofluorescence analysis was performed to assess the expression levels of CYP1B1 and the fibroblast marker PDGF-D in spinal cord tissues from the SHAM, SCI and SCI + Que groups, with n=6. Scale bar, 50 µm. (I) Quantitative fluorescence analysis of CYP1B1 expression was conducted on spinal cord tissues from the SHAM, SCI and SCI + Que groups. (J) RT-qPCR evaluation of CYP1B1 mRNA expression in fibroblasts treated under control conditions, with 10 ng/ml TGF-β and with 10 ng/ml TGF-β + 10 µmol/ml Que, with mRNA levels normalized to GAPDH. (K) Western blot analysis was performed to evaluate CYP1B1 expression in fibroblasts subjected to control conditions, 10 ng/ml TGF-β stimulation or co-treatment with 10 ng/ml TGF-β and 10 µmol/ml Que. (L) Semi-quantitative evaluation of CYP1B1 protein expression in fibroblasts subjected to control conditions, treated with 10 ng/ml TGF-β or treated with 10 ng/ml TGF-β combined with 10 µmol/ml Que, with protein levels normalized to GAPDH. (M) A semi-quantitative analysis was conducted on the Fn protein levels in fibroblasts subjected to control conditions, 10 ng/ml TGF-β treatment or treatment with a combination of 10 ng/ml TGF-β and 10 µmol/ml Que, with the resulting fold-changes normalized against GAPDH. (N) Western blot analysis was utilized to investigate Fn expression in fibroblasts under control conditions, after treatment with 10 ng/ml TGF-β or after co-treatment with 10 ng/ml TGF-β and 10 µmol/ml Que. The data are presented as the mean ± SEM, with sample sizes of n=6 per group for (B, C, D, E, F, G, H and I), n=3 per group for (J) and n=4 per group for (K, L, M and N). *P<0.05, **P<0.01, ***P<0.001 and ****P<0.0001. BMS, Basso Mouse Scale; Con, control; CYP1B1, cytochrome P450 family 1 subfamily B member 1; PDGF-D, platelet-derived growth factor D; dpi, days post-injury; Fn, Fibronectin; MWT, mechanical withdrawal threshold; Que, quercetin; RT-qPCR, reverse transcription-quantitative PCR; SCI, spinal cord injury; SHAM, sham surgery group; TWL, thermal withdrawal latency.

Article Snippet: The protein samples were then transferred to polyvinylidene fluoride membranes, which were subsequently blocked for 1 h at room temperature., washed with TBST solution and incubated with the following primary antibodies: CYP1B1 (1:500; cat. no. 18505-1-AP; Proteintech Group, Inc.), Fibronectin (Fn) (1:1,000; cat. no. ab2413; Abcam) and GAPDH (1:50,000; cat. no. 1E6D9; Proteintech Group, Inc.).

Techniques: Quantitative RT-PCR, Expressing, Western Blot, Immunofluorescence, Marker, Fluorescence, Control, Derivative Assay, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: Molecular Medicine Reports

Article Title: Exploring the role of cytochrome P450 family 1 subfamily B member 1 and quercetin in modulating neuropathic pain after spinal cord injury

doi: 10.3892/mmr.2025.13717

Figure Lengend Snippet: Mechanism diagram. CYP1B1, cytochrome P450 family 1 subfamily B member 1; SCI, spinal cord injury.

Article Snippet: The protein samples were then transferred to polyvinylidene fluoride membranes, which were subsequently blocked for 1 h at room temperature., washed with TBST solution and incubated with the following primary antibodies: CYP1B1 (1:500; cat. no. 18505-1-AP; Proteintech Group, Inc.), Fibronectin (Fn) (1:1,000; cat. no. ab2413; Abcam) and GAPDH (1:50,000; cat. no. 1E6D9; Proteintech Group, Inc.).

Techniques:

Journal: mSystems

Article Title: Quercetin alleviates ulcerative colitis via regulating gut microbiota and tryptophan metabolism

doi: 10.1128/msystems.00703-25

Figure Lengend Snippet: ILA alleviated DSS-induced colitis in mice. ( A ) Flow diagram of the experiment. ( B ) Body weight change (%). ( C ) DAI score. ( D ) Colon image. Histogram statistic of colon length ( E ) and spleen index ( F ). Data were shown as mean ± SEM ( N = 6). ( G ) Representative H&E staining image of colon tissue (scale bar = 200 µm). The relative mRNA expression of TNF-α ( H ) and IL-1β ( I ) in colon tissue was detected by RT-qPCR. ( J–L ) The protein expression of claudin-1 and CYP1B1 was detected by Western blot ( N = 3). * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001 vs the DSS group. ns, not significant.

Article Snippet: After being blocked at room temperature for 1.5 h, four primary antibodies were added successively and incubated at 4°C for 16 h: the tight junction protein Occludin (1:1,000, Proteintech, 66378-1-Ig), Claudin-1 (from the same brand, 1:1,000), Cytochrome P450 1B1 (Proteintech, 18505-1-AP, 1:1,000), and the internal reference GAPDH (Affinity, 13050-1-AP/T0004, 1:10,000).

Techniques: Staining, Expressing, Quantitative RT-PCR, Western Blot

Journal: mSystems

Article Title: Quercetin alleviates ulcerative colitis via regulating gut microbiota and tryptophan metabolism

doi: 10.1128/msystems.00703-25

Figure Lengend Snippet: Qu through ILA-AhR alleviated DSS-induced colitis in mice. ( A ) Flow diagram of the experiment. ( B ) Body weight change (%). ( C ) DAI score. ( D ) Colon image. Histogram statistic of colon length ( E ) and spleen index ( F ). Data were shown as mean ± SEM ( N = 6). ( G ) Representative H&E staining image of colon tissue (scale bar = 200 µm). ( H–J ) The protein expression of claudin-1 and CYP1B1 was detected by western blot ( N = 3). The relative mRNA expression of TNF-α ( K ) and IL-1β ( L ) in colon tissue was detected by RT-qPCR ( N = 3). * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001 vs the DSS group. ns, not significant.

Article Snippet: After being blocked at room temperature for 1.5 h, four primary antibodies were added successively and incubated at 4°C for 16 h: the tight junction protein Occludin (1:1,000, Proteintech, 66378-1-Ig), Claudin-1 (from the same brand, 1:1,000), Cytochrome P450 1B1 (Proteintech, 18505-1-AP, 1:1,000), and the internal reference GAPDH (Affinity, 13050-1-AP/T0004, 1:10,000).

Techniques: Staining, Expressing, Western Blot, Quantitative RT-PCR