Journal: International Journal of Molecular Medicine

Article Title: Apremilast prevents IL-17-induced cellular senescence in ATDC5 chondrocytes mediated by SIRT1

doi: 10.3892/ijmm.2021.4845

Figure Lengend Snippet: Apremilast prevents IL-17-induced expression and secretions of pro-inflammatory cytokines in mouse ATDC5 chondrocytes. Cells were treated with IL-17 (10 ng/ml) in the presence or absence of apremilast (0.5 and 1 µ M) for 24 h. (A) Molecular structure of apremilast. (B) mRNA levels of IL-1β and MCP-1. (C) Secretions of IL-1β and MCP-1. #### P<0.0001 vs. the vehicle control group; ** P<0.01 and *** P<0.001 vs. the IL-17 group. IL, interleukin; MCP-1, monocyte chemoattractant protein-1.

Article Snippet: For the treatment experiment, the cells were plated in 6-well cell culture plates and treated with IL-17 (10 ng/ml) (R&D Systems, Inc.) in the presence or absence of apremilast (0.5 and 1 µ M) (MedChemExpress) at 37°C for 24 h.

Techniques: Expressing, Control

Journal: International Journal of Molecular Medicine

Article Title: Apremilast prevents IL-17-induced cellular senescence in ATDC5 chondrocytes mediated by SIRT1

doi: 10.3892/ijmm.2021.4845

Figure Lengend Snippet: Apremilast prevents IL-17-induced production of ROS in mouse ATDC5 chondrocytes. Cells were treated with IL-17 (10 ng/ml) in the presence or absence of apremilast (0.5 and 1 µ M) for 24 h. Production of ROS was measured using DCFH-DA staining (magnification, ×10). #### P<0.0001 vs. the vehicle control group; ** P<0.01 and *** P<0.001 vs. the IL-17 group. ROS, reactive oxygen species; IL, interleukin; DCFH-DA, 2,7-dichlorodihy-drofluorescein diacetate.

Article Snippet: For the treatment experiment, the cells were plated in 6-well cell culture plates and treated with IL-17 (10 ng/ml) (R&D Systems, Inc.) in the presence or absence of apremilast (0.5 and 1 µ M) (MedChemExpress) at 37°C for 24 h.

Techniques: Staining, Control

Journal: International Journal of Molecular Medicine

Article Title: Apremilast prevents IL-17-induced cellular senescence in ATDC5 chondrocytes mediated by SIRT1

doi: 10.3892/ijmm.2021.4845

Figure Lengend Snippet: Apremilast prevents IL-17-induced cellular senescence in mouse ATDC5 chondrocytes. Cells were treated with IL-17 (10 ng/ml) in the presence or absence of apremilast (1 µ M) for 7 days. Cellular senescence was measured using SA-β-gal staining. ## P<0.01 vs. the vehicle control group; ** P<0.01 vs. the IL-17 group. IL, interleukin; SA-β-gal, senescence-associa ted-β-galactosidase.

Article Snippet: For the treatment experiment, the cells were plated in 6-well cell culture plates and treated with IL-17 (10 ng/ml) (R&D Systems, Inc.) in the presence or absence of apremilast (0.5 and 1 µ M) (MedChemExpress) at 37°C for 24 h.

Techniques: Staining, Control

Journal: International Journal of Molecular Medicine

Article Title: Apremilast prevents IL-17-induced cellular senescence in ATDC5 chondrocytes mediated by SIRT1

doi: 10.3892/ijmm.2021.4845

Figure Lengend Snippet: Apremilast prevents IL-17-induced cell cycle arrest in the G0/G1 phase in mouse ATDC5 chondrocytes. Cells were treated with IL-17 (10 ng/ml) in the presence or absence of apremilast (1 µ M) for 7 days. The cell cycle was assayed using flow cytometry. G0/G1 phase, G2/M phase, and S phase of cells were measured. ## P<0.01 vs. the vehicle control group; ** P<0.01 vs. the IL-17 group. IL, interleukin.

Article Snippet: For the treatment experiment, the cells were plated in 6-well cell culture plates and treated with IL-17 (10 ng/ml) (R&D Systems, Inc.) in the presence or absence of apremilast (0.5 and 1 µ M) (MedChemExpress) at 37°C for 24 h.

Techniques: Flow Cytometry, Control

Journal: International Journal of Molecular Medicine

Article Title: Apremilast prevents IL-17-induced cellular senescence in ATDC5 chondrocytes mediated by SIRT1

doi: 10.3892/ijmm.2021.4845

Figure Lengend Snippet: Apremilast prevents IL-17-induced expression of p21 and PAI-1 in mouse ATDC5 chondrocytes. Cells were treated with IL-17 (10 ng/ml) in the presence or absence of apremilast (1 µ M) for 24 h. (A) mRNA levels of p21 and PAI-1. (B) Protein levels of p21 and PAI-1 as measured by western blotting. #### P<0.0001 vs. the vehicle control group; *** P<0.001 vs. the IL-17 group. IL, interleukin; PAI-1, plasminogen activator inhibitor-1.

Article Snippet: For the treatment experiment, the cells were plated in 6-well cell culture plates and treated with IL-17 (10 ng/ml) (R&D Systems, Inc.) in the presence or absence of apremilast (0.5 and 1 µ M) (MedChemExpress) at 37°C for 24 h.

Techniques: Expressing, Western Blot, Control

Journal: International Journal of Molecular Medicine

Article Title: Apremilast prevents IL-17-induced cellular senescence in ATDC5 chondrocytes mediated by SIRT1

doi: 10.3892/ijmm.2021.4845

Figure Lengend Snippet: Apremilast prevents IL-17-induced reduction of SIRT1 in mouse ATDC5 chondrocytes. Cells were treated with IL-17 (10 ng/ml) in the presence or absence of a Apremilast (1 µ M) for 24 h. (A) mRNA of SIRT1. (B) Protein level of SIRT1. #### P<0.0001 vs. the vehicle control group; *** P<0.001 vs. the IL-17 group. IL, interleukin; SIRT1, sirtuin 1.

Article Snippet: For the treatment experiment, the cells were plated in 6-well cell culture plates and treated with IL-17 (10 ng/ml) (R&D Systems, Inc.) in the presence or absence of apremilast (0.5 and 1 µ M) (MedChemExpress) at 37°C for 24 h.

Techniques: Control

Journal: International Journal of Molecular Medicine

Article Title: Apremilast prevents IL-17-induced cellular senescence in ATDC5 chondrocytes mediated by SIRT1

doi: 10.3892/ijmm.2021.4845

Figure Lengend Snippet: Knockdown of SIRT1 abolishes the protective effects of apremilast against IL-17-induced cellular senescence and the expression levels of p21 and PAI-1. Cells were transfected with SIRT1 siRNA, followed by stimulation with IL-17 (10 ng/ml) in the presence or absence of apremilast (1 µ M) for 24 h. (A) Cellular senescence. (B) mRNA levels of p21 and PAI-1. (C) Protein levels of p21 and PAI-1 as measured by western blotting. #### P<0.0001 vs. the vehicle control group; ****P<0.0001 vs. the IL-17 group; $$$$ P<0.0001 vs. the IL-17 + apremilast group. SIRT1, sirtuin 1; IL, interleukin; PAI-1, plasminogen activator inhibitor-1.

Article Snippet: For the treatment experiment, the cells were plated in 6-well cell culture plates and treated with IL-17 (10 ng/ml) (R&D Systems, Inc.) in the presence or absence of apremilast (0.5 and 1 µ M) (MedChemExpress) at 37°C for 24 h.

Techniques: Knockdown, Expressing, Transfection, Western Blot, Control

Journal: International Journal of Molecular Medicine

Article Title: Apremilast prevents IL-17-induced cellular senescence in ATDC5 chondrocytes mediated by SIRT1

doi: 10.3892/ijmm.2021.4845

Figure Lengend Snippet: Graphical representation of the underlying mechanism, whereby apremilast prevents IL-17-induced cellular senescence in ATDC5 chondrocytes. IL, interleukin; ROS, reactive oxygen species; SIRT1, sirtuin 1; MCP-1, monocyte chemoattractant protein-1; PAI-1, plasminogen activator inhibitor-1.

Article Snippet: For the treatment experiment, the cells were plated in 6-well cell culture plates and treated with IL-17 (10 ng/ml) (R&D Systems, Inc.) in the presence or absence of apremilast (0.5 and 1 µ M) (MedChemExpress) at 37°C for 24 h.

Techniques:

Journal: ACS chemical neuroscience

Article Title: KVA-D-88, a novel preferable phosphodiesterase 4B inhibitor, decreases cocaine-mediated reward properties in vivo

doi: 10.1021/acschemneuro.0c00170

Figure Lengend Snippet: KVA-D-88 was administered into mice (n = 4) by either i.p. injection (10 mg/kg) or p.o. approach (10 mg/kg) and 30 min or 24 hours later, mice were sacrificed for the removal blood and brain. The concentrations of KVA-D-88 were determined in these two tissues to calculate the concentration ratio of brain vs. plasma. (A) Brain and plasma KVA-D-88 concentration 30 min after i.p. injection (n = 4). (B) The IC50 of KVA-D-88 on PDE4B and PDE4D in vitro. HEK293 cells were cultured and transfected with PDE4B1 expression vector, CRE luciferase reporter and a control renilla luciferase vector. The cells were dosed with KVA-D-88 and incubated overnight. Forskolin was added and incubated for 5 – 6 h. A dual luciferase assay was performed for measuring firefly luminescence. The intensity of renilla luminescence was served as internal control. Cell based assays were performed in triplicate at each concentration. The EC50 value was determined by the concentration causing a half-maximal percent activity. (C) The non-linear dose-response curve of KVA-D-88 on PDE4. (D) The non-linear dose-response curve of apremilast on PDE4. (E) The table of EC50 of KVA-D-88 and apremilast on PDE4 in vitro.

Article Snippet: The EC 50 of KVA-D-88 to elevate cAMP levels was 0.5 μM, which is 10-fold lower, compared to the EC 50 of apremilast in vitro (data collected at BPS Biosciences, San Diego, CA).

Techniques: Injection, Concentration Assay, In Vitro, Cell Culture, Transfection, Expressing, Plasmid Preparation, Luciferase, Incubation, Activity Assay

Journal: International Journal of Molecular Sciences

Article Title: Phosphodiesterase-4 Inhibition Reduces Cutaneous Inflammation and IL-1β Expression in a Psoriasiform Mouse Model but Does Not Inhibit Inflammasome Activation

doi: 10.3390/ijms222312878

Figure Lengend Snippet: Heterozygous K5.Stat3C mice were tape-stripped for two weeks to induce psoriatic skin lesions. These mice were then either left untreated or treated with 2 mg/kg/day or 6 mg/kg/day of apremilast for two weeks. Macroscopic appearance ( A ) and histology ( B ) are shown. After Hematoxilin Eosin coloration of skin sections, total leukocytes were counted in 300 µm 2 fields ( E ). Skin sections were then stained with an anti-mouse CD3 ( C ) and with an anti-Ly6G ( D ) antibody and semi-quantified ( F , G ). Expression levels were qualified as very strong (++++), strong (+++), moderate (++), weak (+) or absent (0). Shown are the mean +/− SEM of leukocyte cell counts and CD3+ and Ly6G+ expression levels in 5 mice/condition. ****: p < 0.0001, ***: p < 0.001, *: p < 0.05.

Article Snippet: PBMCs were then primed with upLPS (100 ng/mL) for 16 h. Apremilast or 1 μM Z-VAD (Selleck Chemicals, Houston, TX, USA) treatment was applied for 1 h before stimulation with 1 μM nigericin (Selleck Chemicals, Houston, TX, USA) for PBMCs or with 86.4 mJ/cm 2 UVB (UV802L; Waldmann, Villingen-Schwenningen, Germany), nigericin [ ] or SDS (15 μg/mL) for keratinocytes.

Techniques: Staining, Expressing

Journal: International Journal of Molecular Sciences

Article Title: Phosphodiesterase-4 Inhibition Reduces Cutaneous Inflammation and IL-1β Expression in a Psoriasiform Mouse Model but Does Not Inhibit Inflammasome Activation

doi: 10.3390/ijms222312878

Figure Lengend Snippet: Quantitative PCR was performed on lesional skin samples of heterozygous K5.Stat3C mice either left untreated or treated with 2 mg/kg/day or 6 mg/kg/day apremilast for two weeks. qPCR was performed with primers for the indicated murine cytokines. Non-lesional skin from K5.Stat3C mice served as an internal control (2 − ᴧᴧ CT = 1). Shown is the mean ± SEM of 5 mice/condition. Statistical analysis was performed using One-way Anova followed by Tukey’s posthoc test. ****: p < 0.0001, ***: p < 0.001, **: p < 0.01, *: p < 0.05, ns: p ≥ 0.05.

Article Snippet: PBMCs were then primed with upLPS (100 ng/mL) for 16 h. Apremilast or 1 μM Z-VAD (Selleck Chemicals, Houston, TX, USA) treatment was applied for 1 h before stimulation with 1 μM nigericin (Selleck Chemicals, Houston, TX, USA) for PBMCs or with 86.4 mJ/cm 2 UVB (UV802L; Waldmann, Villingen-Schwenningen, Germany), nigericin [ ] or SDS (15 μg/mL) for keratinocytes.

Techniques: Real-time Polymerase Chain Reaction, Control

Journal: International Journal of Molecular Sciences

Article Title: Phosphodiesterase-4 Inhibition Reduces Cutaneous Inflammation and IL-1β Expression in a Psoriasiform Mouse Model but Does Not Inhibit Inflammasome Activation

doi: 10.3390/ijms222312878

Figure Lengend Snippet: Human PBMCs were treated with 0.1 µM, 1 µM or 10 µM apremilast for one hour followed by upLPS (100 ng/mL) or anti-CD3 stimulation (50 μg/mL). After overnight incubation at 37 °C, cells were harvested for RNA isolation. The mRNA expression levels were detected using qPCR with primers for IL1B, IL6, IL8, IL23, IL17A, IL22, TNFA and IFNG. Shown is the mean ± SEM of 4 healthy individuals. Statistical analysis was performed using One-way Anova followed by Tukey’s posthoc test. ***: p < 0.001, **: p < 0.01, *: p < 0.05, ns: p ≥ 0.05, +: with, -: without.

Article Snippet: PBMCs were then primed with upLPS (100 ng/mL) for 16 h. Apremilast or 1 μM Z-VAD (Selleck Chemicals, Houston, TX, USA) treatment was applied for 1 h before stimulation with 1 μM nigericin (Selleck Chemicals, Houston, TX, USA) for PBMCs or with 86.4 mJ/cm 2 UVB (UV802L; Waldmann, Villingen-Schwenningen, Germany), nigericin [ ] or SDS (15 μg/mL) for keratinocytes.

Techniques: Incubation, Isolation, Expressing

Journal: International Journal of Molecular Sciences

Article Title: Phosphodiesterase-4 Inhibition Reduces Cutaneous Inflammation and IL-1β Expression in a Psoriasiform Mouse Model but Does Not Inhibit Inflammasome Activation

doi: 10.3390/ijms222312878

Figure Lengend Snippet: ( A ) Human PBMCs were stimulated with upLPS overnight, followed by treatment with Z-VAD, or 0.1 µM, 1 µM or 10 µM apremilast for one hour before nigericin was added. ( B ) Cell death was measured using a lactate dehydrogenase (LDH) assay after stimulation of PBMCs with upLPS + Nigericin and apremilast at different doses. ( C ) Human primary keratinocytes were cultured for 24 h before treatment with upLPS overnight, followed by treatment with Z-VAD, or 0.1 µM, 1 µM or 10 µM apremilast two hrs before nigericin or SDS treatment or UVB-irradiation (0.5 J/cm 2 ). After five hrs incubation at 37 °C, the supernatant was collected and IL-1β ELISA was performed. Mean ± SEM of four different healthy individuals is shown.***: p < 0.001, +: with, -: without.

Article Snippet: PBMCs were then primed with upLPS (100 ng/mL) for 16 h. Apremilast or 1 μM Z-VAD (Selleck Chemicals, Houston, TX, USA) treatment was applied for 1 h before stimulation with 1 μM nigericin (Selleck Chemicals, Houston, TX, USA) for PBMCs or with 86.4 mJ/cm 2 UVB (UV802L; Waldmann, Villingen-Schwenningen, Germany), nigericin [ ] or SDS (15 μg/mL) for keratinocytes.

Techniques: Lactate Dehydrogenase Assay, Cell Culture, Irradiation, Incubation, Enzyme-linked Immunosorbent Assay

Journal: Drug Design, Development and Therapy

Article Title: Pharmacokinetics and Bioequivalence of Apremilast Tablets in Chinese Healthy Subjects Under Fasting and Postprandial States

doi: 10.2147/DDDT.S461771

Figure Lengend Snippet: Pharmacokinetic analysis of apremilast formulations during fasting condition. Mean plasma concentration (±SD) time curve after oral test or reference formulation: arithmetic mean ( A ) and log transformation ( B ).

Article Snippet: The test formulation was an apremilast tablet, which was produced and provided by CSPC Ouyi Pharmaceutical Co., Ltd., Hebei, China (30 mg/tablet, batch number: A82190601; expiry date: June 12, 2021).

Techniques: Clinical Proteomics, Concentration Assay, Formulation, Transformation Assay

Journal: Drug Design, Development and Therapy

Article Title: Pharmacokinetics and Bioequivalence of Apremilast Tablets in Chinese Healthy Subjects Under Fasting and Postprandial States

doi: 10.2147/DDDT.S461771

Figure Lengend Snippet: Pharmacokinetic analysis of apremilast formulations during postprandial condition. Mean plasma concentration (±SD) time curve after oral test or reference formulation: arithmetic mean ( A ) and log transformation ( B ).

Article Snippet: The test formulation was an apremilast tablet, which was produced and provided by CSPC Ouyi Pharmaceutical Co., Ltd., Hebei, China (30 mg/tablet, batch number: A82190601; expiry date: June 12, 2021).

Techniques: Clinical Proteomics, Concentration Assay, Formulation, Transformation Assay

Journal: Frontiers in Oncology

Article Title: Pembrolizumab-Induced Psoriasis in Metastatic Melanoma: Activity and Safety of Apremilast, a Case Report

doi: 10.3389/fonc.2020.579445

Figure Lengend Snippet: Clinical presentation of the patient before the treatment with Apremilast (A) . Resolution of psoriasis after the treatment with Apremilast (B) .

Article Snippet: Apremilast (Otezla, Amgen) is a small oral molecule that selectively inhibits PDE-4.

Techniques: