Journal: Frontiers in Pharmacology

Article Title: Menisoxoisoaporphine A, a novel oxoisoaporphine alkaloid from Menispermi Rhizoma, inhibits inflammation by targeting PDE4B

doi: 10.3389/fphar.2024.1505116

Figure Lengend Snippet: Primer sequences used in this study.

Article Snippet: PDE4B-IN-3 (IN-3, purity ≥98%) was purchased from MedChemExpress (Shanghai, China).

Techniques:

Journal: Frontiers in Pharmacology

Article Title: Menisoxoisoaporphine A, a novel oxoisoaporphine alkaloid from Menispermi Rhizoma, inhibits inflammation by targeting PDE4B

doi: 10.3389/fphar.2024.1505116

Figure Lengend Snippet: siRNA sequences used in the study.

Article Snippet: PDE4B-IN-3 (IN-3, purity ≥98%) was purchased from MedChemExpress (Shanghai, China).

Techniques: Sequencing, Control

Journal: Frontiers in Pharmacology

Article Title: Menisoxoisoaporphine A, a novel oxoisoaporphine alkaloid from Menispermi Rhizoma, inhibits inflammation by targeting PDE4B

doi: 10.3389/fphar.2024.1505116

Figure Lengend Snippet: RNA-seq analysis revealed the anti-inflammatory properties of Menisoxoisoaporphine A (MA) were associated with PDE4B. RAW264.7 cells were stimulated with 1 μg·mL -1 lipopolysaccharide (LPS), and concurrently treated with MA (0, 12 μM) for 12 h. The cells were harvested for transcriptome profiling by RNA-seq. (A) Volcano plot showing DEGs between control vs . model groups. (B) Volcano plot showing DEGs between model vs . MA groups. (C) Venn diagram showing the co-differentially expressed genes (co-DEGs) between control vs . model and model vs . MA groups. (D) Heatmap showing the expression of 117 co-DEGs. (E) The mRNA relative expression of Pde4b, Igf2r, Slc6a12, Itgb7, Tlr9, Nox1, Mmp12, Abca1, Cd80, Il11 and Tnfrsf9 were validated by qRT-PCR in RAW264.7 cells. For the volcano plot, the abscissa represents the Log2 transformed fold-change, and the ordinate represents the -log10 transformed p -value. The green dots indicated the downregulated genes, the red dots indicated the upregulated genes and the grey dots indicated the non-DEGs. DEGs: differentially expressed genes (FPKM value, fold change ≥1.5 and p ≤ 0.05). Control (C1-C3): control normal group; Model (M1-M3): only LPS-treated group; MA (MA1-MA3): LPS and MA-treated group. Each value was expressed as the means ± SEM (n = 3). # p < 0.05: vs . control group. * p < 0.05: vs . only LPS-treated group.

Article Snippet: PDE4B-IN-3 (IN-3, purity ≥98%) was purchased from MedChemExpress (Shanghai, China).

Techniques: RNA Sequencing Assay, Control, Expressing, Quantitative RT-PCR, Transformation Assay

Journal: Frontiers in Pharmacology

Article Title: Menisoxoisoaporphine A, a novel oxoisoaporphine alkaloid from Menispermi Rhizoma, inhibits inflammation by targeting PDE4B

doi: 10.3389/fphar.2024.1505116

Figure Lengend Snippet: Menisoxoisoaporphine A (MA) inhibited LPS-induced inflammation via PDE4B-cAMP-PKA-NF-κB pathway. RAW264.7 cells were stimulated with 1 μg·mL -1 lipopolysaccharide (LPS), and concurrently treated with MA (0, 12 μM) or PDE4B-IN-3 (IN-3, 1 μM) for 12 h. (A) Representative Western blot bands and (B–F) quantification of PDE4B, cAMP, p-PKA, p-P65(Ser276)/P65 and p-IκB(Ser32/36)/IκB. β-Actin was used as an internal reference. Each value was expressed as the means ± SEM (n = 3). # p < 0.05: vs . control group. * p < 0.05: vs . only LPS-treated group.

Article Snippet: PDE4B-IN-3 (IN-3, purity ≥98%) was purchased from MedChemExpress (Shanghai, China).

Techniques: Western Blot, Control

Journal: Frontiers in Pharmacology

Article Title: Menisoxoisoaporphine A, a novel oxoisoaporphine alkaloid from Menispermi Rhizoma, inhibits inflammation by targeting PDE4B

doi: 10.3389/fphar.2024.1505116

Figure Lengend Snippet: PDE4B knockdown abolished the anti-inflammatory effects of Menisoxoisoaporphine A (MA) at LPS-induced RAW264.7 cells. RAW264.7 cells were transfected with Pde4b siRNA or control siRNA for 12 h. (A) The PDE4B proteins were probed by western bolt analysis. Transfected cells were then stimulated with 1 μg·mL −1 lipopolysaccharide (LPS), and concurrently treated with MA (0, 12 μM) for 12 h. (B) Representative Western blot banding pictures and quantification of PDE4B, cAMP, p-PKA, p-IκB(Ser32/36)/IκB and p-P65(Ser276)/P65 of Pde4b siRNA#2 treatment. (C) Representative Western blot banding pictures and quantification of PDE4B, cAMP, p-PKA, p-IκB(Ser32/36)/IκB and p-P65(Ser276)/P65 of Pde4b siRNA#4 treatment. β-Actin was used as an internal reference. NC, cells without any treatment. SC, cells only treated with control siRNA. Each value was expressed as the means ± SEM (n = 3). # p < 0.05: vs . control siRNA group. * p < 0.05: vs . indicated group.

Article Snippet: PDE4B-IN-3 (IN-3, purity ≥98%) was purchased from MedChemExpress (Shanghai, China).

Techniques: Knockdown, Transfection, Control, Western Blot

Journal: Frontiers in Pharmacology

Article Title: Menisoxoisoaporphine A, a novel oxoisoaporphine alkaloid from Menispermi Rhizoma, inhibits inflammation by targeting PDE4B

doi: 10.3389/fphar.2024.1505116

Figure Lengend Snippet: Menisoxoisoaporphine A (MA) inhibited cAMP/PKA-NF-κB via PDE4B. RAW264.7 cells were transfected with pIRES2-EGFP-Pde4b overexpressed plasmid or empty pIRES2-EGFP for 12 h. (A) PDE4B protein expression detected by western bolt. Transfected cells were then stimulated with 1 μg·mL −1 lipopolysaccharide (LPS), and concurrently treated with MA (0, 12 μM) for 12 h. (B) Representative Western blot bands and quantification of PDE4B, cAMP, p-PKA, p-P65(Ser276)/P65 and p-IκB(Ser32/36)/IκB. β-Actin was used as an internal reference. Each value was expressed as the means ± SEM (n = 3). # p < 0.05: vs . control hGFP group. * p < 0.05: vs . indicated group.

Article Snippet: PDE4B-IN-3 (IN-3, purity ≥98%) was purchased from MedChemExpress (Shanghai, China).

Techniques: Transfection, Plasmid Preparation, Expressing, Western Blot, Control

Journal: Frontiers in Pharmacology

Article Title: Menisoxoisoaporphine A, a novel oxoisoaporphine alkaloid from Menispermi Rhizoma, inhibits inflammation by targeting PDE4B

doi: 10.3389/fphar.2024.1505116

Figure Lengend Snippet: Menisoxoisoaporphine A (MA) could directly bind to PDE4B at Tyr405 site. (A) Molecular docking simulation between MA and PDE4B. (B) The root mean square deviation (RMSD) of PDE4B in the PDE4B WT system and PDE4B Y405A system. (C) The RMSD of MA in the PDE4B WT system and PDE4B Y405A system. (D) The residual energy decomposition of the binding free energy of PDE4B WT system and PDE4B Y405A system, the horizontal axis denotes the respective amino acids associated with each position. WT: wild type; TYR: tyrosine; Y405A: The tyrosine residue at position 405 is mutated to glycine.

Article Snippet: PDE4B-IN-3 (IN-3, purity ≥98%) was purchased from MedChemExpress (Shanghai, China).

Techniques: Binding Assay, Residue

Journal: Frontiers in Pharmacology

Article Title: Menisoxoisoaporphine A, a novel oxoisoaporphine alkaloid from Menispermi Rhizoma, inhibits inflammation by targeting PDE4B

doi: 10.3389/fphar.2024.1505116

Figure Lengend Snippet: Binding free energy and different energy contributions of MA-PDE4B WT systems and MA-PDE4B Y405A systems (kcal/mol).

Article Snippet: PDE4B-IN-3 (IN-3, purity ≥98%) was purchased from MedChemExpress (Shanghai, China).

Techniques: Binding Assay

Journal: Frontiers in Pharmacology

Article Title: Menisoxoisoaporphine A, a novel oxoisoaporphine alkaloid from Menispermi Rhizoma, inhibits inflammation by targeting PDE4B

doi: 10.3389/fphar.2024.1505116

Figure Lengend Snippet: Menisoxoisoaporphine A (MA) inhibited cAMP-PKA-NF-κB mediated inflammation via targeting PDE4B.

Article Snippet: PDE4B-IN-3 (IN-3, purity ≥98%) was purchased from MedChemExpress (Shanghai, China).

Techniques: