pde4a enzymatic activity  (BPS Bioscience)

Journal: Frontiers in Pharmacology

Article Title: Arecoline Enhances Phosphodiesterase 4A Activity to Promote Transforming Growth Factor-β-Induced Buccal Mucosal Fibroblast Activation via cAMP-Epac1 Signaling Pathway

doi: 10.3389/fphar.2021.722040

Figure Lengend Snippet: Identification of proteins interacting with arecoline by bioinformatics and experimental analyses. (A,B) KEGG signaling pathway annotation and GO target genes function significance analyses were performed on candidate proteins that might interact with arecoline. (C) The mRNA levels of PDE4A, PDE6D and PDE7A in collected normal oral mucosal and OSF tissues. (D) BMFs were treated with 5 ng/ml TGF-β1 for 0, 24, and 48 h and examined for the protein levels of PDE4A by Immunoblotting, n = 3. (E) cAMP concentration was determined by a direct cAMP ELISA kit. n = 3, ** p < 0.01 compared with the normal or control group. ANOVA followed by Tukey post-hoc test.

Article Snippet: PDE4A enzymatic activity was determined using a PDE4A assay kit (Cat. 60,340, BPS Bioscience Inc., United States).

Techniques: Western Blot, Concentration Assay, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Pharmacology

Article Title: Arecoline Enhances Phosphodiesterase 4A Activity to Promote Transforming Growth Factor-β-Induced Buccal Mucosal Fibroblast Activation via cAMP-Epac1 Signaling Pathway

doi: 10.3389/fphar.2021.722040

Figure Lengend Snippet: Arecoline enhances TGF-β1-induced BMFs activation. BMFs were transfected with si-PDE4A in the presence or absence of 50 µg/ml arecoline treatment upon 5 ng/ml TGF-β1 stimulation for 48 h, and then examined for (A) the protein levels of PDE4A by Immunoblotting, n = 3. (B) the cAMP concentration by ELISA, n = 3. (C) the protein levels of p-Smad2, α-SMA, and Col1A1 by Immunoblotting, n = 3. (D) the gel contraction capacity, n = 3. (E,F) the migration capacity by Wound healing and Transwell assays, n = 3. ** p < 0.01, compared to control group; # p < 0.05, ## p < 0.01, compared to arecoline group. ANOVA followed by Tukey post-hoc test.

Article Snippet: PDE4A enzymatic activity was determined using a PDE4A assay kit (Cat. 60,340, BPS Bioscience Inc., United States).

Techniques: Activation Assay, Transfection, Western Blot, Concentration Assay, Enzyme-linked Immunosorbent Assay, Migration

Journal: Frontiers in Pharmacology

Article Title: Arecoline Enhances Phosphodiesterase 4A Activity to Promote Transforming Growth Factor-β-Induced Buccal Mucosal Fibroblast Activation via cAMP-Epac1 Signaling Pathway

doi: 10.3389/fphar.2021.722040

Figure Lengend Snippet: Arecoline regulates TGF-β1-induced BMFs activation by affecting the activity of PDE4A. BMFs were co-treated with 50 µg/ml arecoline and 5 ng/ml TGF-β1 in the presence or absence of PDE4 specific inhibitor Rolipram, and then examined for (A) the protein levels of p -Smad2, α-SMA, and Col1A1, n = 3; (B) the gel contraction capacity, n = 3. (C,D) the migration capacity by Wound healing and Transwell assays, n = 3 ** p < 0.01 compared with the control group, student t test.

Article Snippet: PDE4A enzymatic activity was determined using a PDE4A assay kit (Cat. 60,340, BPS Bioscience Inc., United States).

Techniques: Activation Assay, Activity Assay, Migration

Journal: Frontiers in Pharmacology

Article Title: Arecoline Enhances Phosphodiesterase 4A Activity to Promote Transforming Growth Factor-β-Induced Buccal Mucosal Fibroblast Activation via cAMP-Epac1 Signaling Pathway

doi: 10.3389/fphar.2021.722040

Figure Lengend Snippet: Arecoline affects the effects of PDE4A on the cAMP pathway upon TGF-β1 stimulation. BMFs were transfected with si-PDE4A in the presence or absence of 50 µg/ml arecoline treatment upon 5 ng/ml TGF-β1 stimulation, and then examined for (A) the protein levels of PKA, p-PKA, CREB, and p-CREB by Immunoblotting, n = 3. (B) the protein levels of GTP-Rap1, total Rap1, and Epac1 by Immunoblotting, n = 3. ** p < 0.01, compared to the control group; ## p < 0.01, compared to arecoline group. ANOVA followed by Tukey post-hoc test. (C) BMFs were co-treated with arecoline and TGF-β1 in the presence or absence of PKA-selective cAMP analog N6-cAMP, and then examined for the protein levels of Epac1, α-SMA and Col1A1 by Immunoblotting, n = 3. (D) BMFs were co-treated with arecoline and TGF-β1 in the presence or absence of Epac1-selective cAMP analog 8-Me-cAMP, and then examined for the protein levels of Epac1, α-SMA and Col1A1 by Immunoblotting, n = 3. ** p < 0.01, compared with the control group, student t -test.

Article Snippet: PDE4A enzymatic activity was determined using a PDE4A assay kit (Cat. 60,340, BPS Bioscience Inc., United States).

Techniques: Transfection, Western Blot

Journal: Frontiers in Pharmacology

Article Title: Arecoline Enhances Phosphodiesterase 4A Activity to Promote Transforming Growth Factor-β-Induced Buccal Mucosal Fibroblast Activation via cAMP-Epac1 Signaling Pathway

doi: 10.3389/fphar.2021.722040

Figure Lengend Snippet: A schematic diagram. In TGF-β activated buccal mucosal fibroblast, arecoline increased the PDE4A activity, therefore inhibiting the cAMP/Epac1 pathway and promoting the oral submucous fibrosis.

Article Snippet: PDE4A enzymatic activity was determined using a PDE4A assay kit (Cat. 60,340, BPS Bioscience Inc., United States).

Techniques: Activity Assay

Journal: Frontiers in Pharmacology

Article Title: Arecoline Enhances Phosphodiesterase 4A Activity to Promote Transforming Growth Factor-β-Induced Buccal Mucosal Fibroblast Activation via cAMP-Epac1 Signaling Pathway

doi: 10.3389/fphar.2021.722040

Figure Lengend Snippet: Identification of proteins interacting with arecoline by bioinformatics and experimental analyses. (A,B) KEGG signaling pathway annotation and GO target genes function significance analyses were performed on candidate proteins that might interact with arecoline. (C) The mRNA levels of PDE4A, PDE6D and PDE7A in collected normal oral mucosal and OSF tissues. (D) BMFs were treated with 5 ng/ml TGF-β1 for 0, 24, and 48 h and examined for the protein levels of PDE4A by Immunoblotting, n = 3. (E) cAMP concentration was determined by a direct cAMP ELISA kit. n = 3, ** p < 0.01 compared with the normal or control group. ANOVA followed by Tukey post-hoc test.

Article Snippet: PDE4A enzymatic activity was determined using a PDE4A assay kit (Cat. 60,340, BPS Bioscience Inc., United States).

Techniques: Western Blot, Concentration Assay, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Pharmacology

Article Title: Arecoline Enhances Phosphodiesterase 4A Activity to Promote Transforming Growth Factor-β-Induced Buccal Mucosal Fibroblast Activation via cAMP-Epac1 Signaling Pathway

doi: 10.3389/fphar.2021.722040

Figure Lengend Snippet: Arecoline enhances TGF-β1-induced BMFs activation. BMFs were transfected with si-PDE4A in the presence or absence of 50 µg/ml arecoline treatment upon 5 ng/ml TGF-β1 stimulation for 48 h, and then examined for (A) the protein levels of PDE4A by Immunoblotting, n = 3. (B) the cAMP concentration by ELISA, n = 3. (C) the protein levels of p-Smad2, α-SMA, and Col1A1 by Immunoblotting, n = 3. (D) the gel contraction capacity, n = 3. (E,F) the migration capacity by Wound healing and Transwell assays, n = 3. ** p < 0.01, compared to control group; # p < 0.05, ## p < 0.01, compared to arecoline group. ANOVA followed by Tukey post-hoc test.

Article Snippet: PDE4A enzymatic activity was determined using a PDE4A assay kit (Cat. 60,340, BPS Bioscience Inc., United States).

Techniques: Activation Assay, Transfection, Western Blot, Concentration Assay, Enzyme-linked Immunosorbent Assay, Migration

Journal: Frontiers in Pharmacology

Article Title: Arecoline Enhances Phosphodiesterase 4A Activity to Promote Transforming Growth Factor-β-Induced Buccal Mucosal Fibroblast Activation via cAMP-Epac1 Signaling Pathway

doi: 10.3389/fphar.2021.722040

Figure Lengend Snippet: Arecoline regulates TGF-β1-induced BMFs activation by affecting the activity of PDE4A. BMFs were co-treated with 50 µg/ml arecoline and 5 ng/ml TGF-β1 in the presence or absence of PDE4 specific inhibitor Rolipram, and then examined for (A) the protein levels of p -Smad2, α-SMA, and Col1A1, n = 3; (B) the gel contraction capacity, n = 3. (C,D) the migration capacity by Wound healing and Transwell assays, n = 3 ** p < 0.01 compared with the control group, student t test.

Article Snippet: PDE4A enzymatic activity was determined using a PDE4A assay kit (Cat. 60,340, BPS Bioscience Inc., United States).

Techniques: Activation Assay, Activity Assay, Migration

Journal: Frontiers in Pharmacology

Article Title: Arecoline Enhances Phosphodiesterase 4A Activity to Promote Transforming Growth Factor-β-Induced Buccal Mucosal Fibroblast Activation via cAMP-Epac1 Signaling Pathway

doi: 10.3389/fphar.2021.722040

Figure Lengend Snippet: Arecoline affects the effects of PDE4A on the cAMP pathway upon TGF-β1 stimulation. BMFs were transfected with si-PDE4A in the presence or absence of 50 µg/ml arecoline treatment upon 5 ng/ml TGF-β1 stimulation, and then examined for (A) the protein levels of PKA, p-PKA, CREB, and p-CREB by Immunoblotting, n = 3. (B) the protein levels of GTP-Rap1, total Rap1, and Epac1 by Immunoblotting, n = 3. ** p < 0.01, compared to the control group; ## p < 0.01, compared to arecoline group. ANOVA followed by Tukey post-hoc test. (C) BMFs were co-treated with arecoline and TGF-β1 in the presence or absence of PKA-selective cAMP analog N6-cAMP, and then examined for the protein levels of Epac1, α-SMA and Col1A1 by Immunoblotting, n = 3. (D) BMFs were co-treated with arecoline and TGF-β1 in the presence or absence of Epac1-selective cAMP analog 8-Me-cAMP, and then examined for the protein levels of Epac1, α-SMA and Col1A1 by Immunoblotting, n = 3. ** p < 0.01, compared with the control group, student t -test.

Article Snippet: PDE4A enzymatic activity was determined using a PDE4A assay kit (Cat. 60,340, BPS Bioscience Inc., United States).

Techniques: Transfection, Western Blot

Journal: Frontiers in Pharmacology

Article Title: Arecoline Enhances Phosphodiesterase 4A Activity to Promote Transforming Growth Factor-β-Induced Buccal Mucosal Fibroblast Activation via cAMP-Epac1 Signaling Pathway

doi: 10.3389/fphar.2021.722040

Figure Lengend Snippet: A schematic diagram. In TGF-β activated buccal mucosal fibroblast, arecoline increased the PDE4A activity, therefore inhibiting the cAMP/Epac1 pathway and promoting the oral submucous fibrosis.

Article Snippet: PDE4A enzymatic activity was determined using a PDE4A assay kit (Cat. 60,340, BPS Bioscience Inc., United States).

Techniques: Activity Assay