ph797804  (MedChemExpress)

Journal: BMC cancer

Article Title: Homoharringtonine enhances cytarabine-induced apoptosis in acute myeloid leukaemia by regulating the p38 MAPK/H2AX/Mcl-1 axis.

doi: 10.1186/s12885-024-12286-7

Figure Lengend Snippet: Fig. 3 HHT induced apoptosis by p38 MAPK/H2AX/Mcl-1 axis. A, B, D HL-60 and THP-1 cells were pretreated with 10 μM SP600125, a p38 inhibitor (10 μM SB203580) or PH797804, an ERK inhibitor (10 μM PD98059), for 4 h and then treated with increasing concentrations of HHT for another 24 h, after which apoptosis was detected by AO-EB staining. C, E, Protein expression in HL-60 or THP-1 cells treated with SB203580 and HHT for 24 h. F, G, p38 was silenced with p38 siRNA in THP-1 cells that were subsequently treated with HHT. *, P < 0.05; **, P < 0.01; ***, P < 0.001

Article Snippet: Cytarabine and PH797804 were obtained from Selleck Chemicals (Houston, TX, USA).

Techniques: Staining, Expressing

Journal: Signal transduction and targeted therapy

Article Title: SARS-CoV-2 spike protein-ACE2 interaction increases carbohydrate sulfotransferases and reduces N-acetylgalactosamine-4-sulfatase by p38 MAPK.

doi: 10.1038/s41392-024-01741-3

Figure Lengend Snippet: Fig. 2 SMAD3 and p38 MAPK inhibitors abrogate the effects of SPRBD. a SIS3, a specific inhibitor of phospho-SMAD3, inhibited the SPRBD- and SPRBD + IFN-β- induced increases in expression of CHST15 and CHST11. b Following silencing of ACE2 by siRNA, the SPRBD- and SPRBD + IFN-β- induced increases in phospho-(Thr180/Tyr182)-p38 MAPK were inhibited. c The SPRBD- and SPRBD + IFN-β- induced increases in phospho-(S423/S425)-SMAD3 were completely inhibited by SIS3 and, to a lesser extent, by SB, the p38 MAPK inhibitor. d The SPRBD and SPRBD + IFN-β- induced increases in phospho-(Thr180/Tyr182)-p38 were unaffected by exposure to SIS3 and were inhibited by SB. e, f Promoter activity of CHST15 and CHST11 was enhanced by exposure to SPRBD and to a greater extent by the combination of IFN-β and SPRBD. Both SIS3 and SB inhibited promoter activation. g Treatment of cells with the specific phospho-p38α/β inhibitor PH797804 or with p38α siRNA completely blocked the SPRBD-induced increase in phospho-p38 MAPK detected by ELISA. In contrast, TAB1 siRNA and control siRNA had no effect. h The SPRBD-induced increases in expression of CHST15 and of CHST11 were inhibited by p38α siRNA and by PH797804, but not by TAB1 siRNA or control siRNA. All p-values were determined by unpaired t test, two-tailed, with unequal variance, and with at least three independent experiments. Error bars represent one standard deviation. *** represents p ≤0.001; **** is for p ≤0.0001. ACE angiotensin- converting enzyme, CHST carbohydrate sulfotransferase, NSC NSC23766, SB SB203580, si siRNA, SIS3 specific inhibitor of SMAD3, SMAD Mothers against decapentaplegic homolog 3 (DPC3), SPRBD spike protein receptor-binding domain, TAB1 TGF-beta activated kinase (MAP3K7)

Article Snippet: Inhibitors included: p38 MAPK Inhibitor SB203580 (SB; Adezmapimod, Tocris Bioscience, Bio-Techne, St. Minneapolis, MN, USA; 10 μM); p38α/p38β inhibitor PH797804 (Selleck Chemicals, Houston, TX, USA; 10 μM); Specific Inhibitor of SMAD3 (SIS3, Tocris, Bio Techne Corporation, Minneapolis, MN, USA; 3 μM); Rho/Rac1 inhibitor NSC23766 (NSC; Tocris; 1 ng/ml); Desloratadine (Sigma-Millipore, St. Louis, MO, USA; 16 μM, 32 μM, 64 μM), a histamine H1 receptor antagonist, reported to activate p38-MAPK;36 and Monensin (Sigma-Millipore; 0.5 μM, 1 μM, 2 μM), a polyether antibiotic which inhibits synthesis of dermatan sulfate and chondroitin sulfate.42–46 The cells were then treated for two hours with purified recombinant SARS-CoV-2 Spike Protein-Receptor Binding Domain protein (SPRBD; AA: Lys310-Leu560; QHD43416; Amsbio, Cambridge, MA, USA; TP750182, 2.5 μg/ml), which was expressed in E.coli cells.

Techniques: Expressing, Activity Assay, Activation Assay, Enzyme-linked Immunosorbent Assay, Control, Two Tailed Test, Standard Deviation, Binding Assay

Journal: Signal transduction and targeted therapy

Article Title: SARS-CoV-2 spike protein-ACE2 interaction increases carbohydrate sulfotransferases and reduces N-acetylgalactosamine-4-sulfatase by p38 MAPK.

doi: 10.1038/s41392-024-01741-3

Figure Lengend Snippet: Fig. 4 Effects of desloratadine, monensin, and carrageenan on expression of CHST15, CHST11, and ARSB. a Exposure to the antihistamine desloratadine at a concentration of 32 µM for 2 h reduced by 62% the SPRBD-induced increase in phospho-(Thr180/Tyr182)-p38 MAPK in the AEC. b Consistent with the decline in phospho-p38 MAPK, desloratadine reduced the SPRBD- and SPRBD + IFN-β- induced increases in CHST15 (p = 0.04; p ≤0.0003 with IFN-β) and CHST11 expression (p = 0.0008; p ≤0.0009 with IFN-β, n = 6). Declines increased with increasing concentrations of desloratadine from 16 µM to 32 µM to 64 µM, without evidence of cytotoxicity. c Fold-change of ARSB mRNA expression increased significantly and progressively with desloratadine 16–64 µM for 2 h (p = 0.02, p = 1.35 × 10−6, p = 4.3 × 10−8 with IFN-β; n = 6). d Desloratadine 32 µM for 2 h partially reversed the SPRBD- and SPRBD + IFN-β- induced declines in ARSB activity (from 64.7 to 73.4 nmol/mg protein/h and from 41.0 to 61.8 nmol/mg protein/h with IFN-β; n = 3) (p = 0.036, p = 0.0042, n = 3). e The polyether antibiotic monensin 1.0 µM for 2 h reduced the SPRBD-induced increases in CHST15 mRNA (p = 0.005; p ≤0.0001 with IFN-β) and CHST11 (p = 0.002, p ≤0.0002 with IFN-β; fold-change compared to control; n = 6). f Monensin had no significant impact on the SPRBD-induced decline in ARSB. g Carrageenan-exposed C57BL/6 J mice had marked increase in phospho-p38 MAPK in lung tissue (p = 6.0 × 10−6, n = 6 per group). h mRNA expression of CHST15 and CHST11 was significantly increased in the lung tissue of the carrageenan-exposed mice (p < 1 × 10−5, n = 6). In contrast, ARSB expression declined significantly (p = 8.4 × 10−9, n = 6). i Consistent with the phospho-38 MAPK-Rb-E2F1-mediated mechanism of regulation of ARSB mRNA expression, the DNA-bound E2F was significantly reduced (p = 1.2 × 10−6, n = 6) in the lung tissue of the carrageenan-exposed mice. All p values were determined using unpaired t tests, two-tailed, with unequal variance, and error bars represent one standard deviation. * represents p ≤0.05; ** is for p ≤0.01, *** for p ≤0.001, and **** for p ≤0.0001. ARSB arylsulfatase B = N- acetylgalactosamine-4-sulfatase, CHST carbohydrate sulfotransferase, Des Desloratadine, IFN interferon, Mon Monensin, ND no difference, PAPSS 3’-phosphoadenosine 5’-phosphosulfate synthase, sGAG sulfated glycosaminoglycan, SPRBD spike protein receptor-binding domain

Article Snippet: Inhibitors included: p38 MAPK Inhibitor SB203580 (SB; Adezmapimod, Tocris Bioscience, Bio-Techne, St. Minneapolis, MN, USA; 10 μM); p38α/p38β inhibitor PH797804 (Selleck Chemicals, Houston, TX, USA; 10 μM); Specific Inhibitor of SMAD3 (SIS3, Tocris, Bio Techne Corporation, Minneapolis, MN, USA; 3 μM); Rho/Rac1 inhibitor NSC23766 (NSC; Tocris; 1 ng/ml); Desloratadine (Sigma-Millipore, St. Louis, MO, USA; 16 μM, 32 μM, 64 μM), a histamine H1 receptor antagonist, reported to activate p38-MAPK;36 and Monensin (Sigma-Millipore; 0.5 μM, 1 μM, 2 μM), a polyether antibiotic which inhibits synthesis of dermatan sulfate and chondroitin sulfate.42–46 The cells were then treated for two hours with purified recombinant SARS-CoV-2 Spike Protein-Receptor Binding Domain protein (SPRBD; AA: Lys310-Leu560; QHD43416; Amsbio, Cambridge, MA, USA; TP750182, 2.5 μg/ml), which was expressed in E.coli cells.

Techniques: Expressing, Concentration Assay, Activity Assay, Control, Two Tailed Test, Standard Deviation, Binding Assay

Journal: Signal transduction and targeted therapy

Article Title: SARS-CoV-2 spike protein-ACE2 interaction increases carbohydrate sulfotransferases and reduces N-acetylgalactosamine-4-sulfatase by p38 MAPK.

doi: 10.1038/s41392-024-01741-3

Figure Lengend Snippet: Fig. 5 SARS-CoV-2 spike protein binding with ACE2 initiates transcriptional events which modify chondroitin sulfation through activation of phospho-p38 MAPK. This schematic presents the signaling mechanism by which the SARS-CoV-2 spike protein receptor-binding domain (SPRBD) interacts and initiates the transcriptional events which lead to increased chondroitin sulfate in airway epithelial cells and in Covid-19 infected lung.1 The activation of phospho-p38 MAPK and phospho-SMAD3 lead to increased expression of CHST15 and CHST11. Phospho-p38 MAPK leads to N-terminal phosphorylation of Rb and enhanced Rb binding with E2F1, which reduces ARSB promoter binding with E2F1 and reduces ARSB expression. Additional effects may arise from ACE2-SPRBD binding, due to reduced availability of ACE2 for other biochemical reactions, including the conversion of AngII to Ang1-7. AngII interaction with the angiotensin II receptor type 1 (AT1R) increases phospho-p38 MAPK, and increased availability of AngII may contribute to the observed downstream effects of p38 MAPK on transcription. The altered balance between AngII-induced vasoconstriction and Ang1-7 vasodilatation, following interaction with the G-protein coupled Mas receptor, may have additional pathophysiological consequences. ACE2 angiotensin-converting enzyme 2, AngII angiotensin II, ARSB arylsulfatase B, AT1R angiotensin II receptor type 1, AT2R angiotensin II receptor type 2, CHST carbohydrate sulfotransferase, Mas Ang1-7 receptor, pRb retinoblastoma protein, SMAD Suppressor of Mothers against Decapentaplegic, SPRBD SARS-CoV-2 spike protein receptor binding domain

Article Snippet: Inhibitors included: p38 MAPK Inhibitor SB203580 (SB; Adezmapimod, Tocris Bioscience, Bio-Techne, St. Minneapolis, MN, USA; 10 μM); p38α/p38β inhibitor PH797804 (Selleck Chemicals, Houston, TX, USA; 10 μM); Specific Inhibitor of SMAD3 (SIS3, Tocris, Bio Techne Corporation, Minneapolis, MN, USA; 3 μM); Rho/Rac1 inhibitor NSC23766 (NSC; Tocris; 1 ng/ml); Desloratadine (Sigma-Millipore, St. Louis, MO, USA; 16 μM, 32 μM, 64 μM), a histamine H1 receptor antagonist, reported to activate p38-MAPK;36 and Monensin (Sigma-Millipore; 0.5 μM, 1 μM, 2 μM), a polyether antibiotic which inhibits synthesis of dermatan sulfate and chondroitin sulfate.42–46 The cells were then treated for two hours with purified recombinant SARS-CoV-2 Spike Protein-Receptor Binding Domain protein (SPRBD; AA: Lys310-Leu560; QHD43416; Amsbio, Cambridge, MA, USA; TP750182, 2.5 μg/ml), which was expressed in E.coli cells.

Techniques: Protein Binding, Activation Assay, Binding Assay, Infection, Expressing, Phospho-proteomics