The Protein Tyrosine Kinase Inhibitor Genistein Suppresses Hypoxia-Induced Atrial Natriuretic Peptide Secretion Mediated by the PI3K/Akt-HIF-1α Pathway in Isolated Beating Rat Atria

Abstract Background Genistein, an isoflavonoid that can inhibit protein tyrosine kinase (PTK) phosphorylation, was proved to play pivotal roles in the signal transduction pathways of hypoxic disorders. Aim of the stud y: In this study, we established a rat model of isolated beating atrium and investigated the regulator role of genistein and its downstream signaling pathways in acute hypoxia-induced ANP secretion. Methods Radio-immunoassay was used to detect the ANP content in the atrial perfusates. Western blot analysis was used to determine the protein level of hypoxia-inducible factor-1α (HIF-1α), and GATA4 in the atrial tissue. Results The results showed that acute hypoxia substantially promoted ANP secretion, whereas this effect was partly attenuated by the PTKs inhibitor genistein (3 µM). By western blotting analysis, we found that hypoxia-induced the increase in phosphorylation of Akt and transcriptional factors, including HIF-1α, were also reversed by genistein. The perfused HIF-1α inhibitors rotenone (0.5 µM) or CAY10585 (10 µM) plus genistein significantly abolished the enhanced ANP section induced by hypoxia. Additionally, the perfused PI3K/Akt agonist IGF-1 (30 µM) also abolished ANP secretion induced by genistein as well as inhibited expression of HIF-1α. Conclusions In summary, our data suggested that acute hypoxia markedly increased ANP secretion by PTKs through the PI3K/HIF-1α depended pathway.

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The protein tyrosine kinase inhibitor genistein suppresses hypoxia-induced atrial natriuretic peptide secretion mediated by the PI3K/Akt-HIF-1α pathway in isolated beating rat atria

Canadian Journal of Physiology and Pharmacology ◽

10.1139/cjpp-2020-0503 ◽

2021 ◽

pp. 1-7

Author(s):

Qiu-li Zhang ◽

Ping Li ◽

Lan Hong ◽

Rui-zhuang Li ◽

Jia-qi Wang ◽

...

Keyword(s):

Atrial Natriuretic Peptide ◽

Tyrosine Kinase ◽

Natriuretic Peptide ◽

Protein Tyrosine Kinase ◽

Kinase Inhibitor ◽

Acute Hypoxia ◽

Hypoxia Inducible Factor ◽

Protein Tyrosine ◽

Downstream Signaling ◽

Atrial Natriuretic

Genistein, an isoflavonoid that can inhibit protein tyrosine kinase (PTK) phosphorylation, has been shown to play pivotal roles in the signal transduction pathways of hypoxic disorders. In this study, we established a rat model of isolated beating atrium and investigated the regulator role of genistein and its downstream signaling pathways in acute hypoxia-induced atrial natriuretic peptide (ANP) secretion. Radioimmunoassay was used to detect the ANP content in the atrial perfusates. Western blot analysis was used to determine the protein level of hypoxia-inducible factor 1α (HIF-1α), and GATA4 in the atrial tissue. The results showed that acute hypoxia substantially promoted ANP secretion, whereas this effect was partly attenuated by the PTKs inhibitor genistein (3 μM). By Western blotting analysis, we found that hypoxia-induced increase in phosphorylation of Akt and transcriptional factors, including HIF-1α, were also reversed by genistein. The perfused HIF-1α inhibitors rotenone (0.5 μM) or CAY10585 (10 μM) plus genistein significantly abolished the enhanced ANP section induced by hypoxia. Additionally, the perfused PI3K/Akt agonist insulin-like growth factor 1 (30 μM) also abolished ANP secretion induced by genistein and inhibited expression of HIF-1α. In summary, our data suggested that acute hypoxia markedly increased ANP secretion by PTKs through the phosphoinositide-3 kinase (PI3K)/HIF-1α dependent pathway.

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Role of Protein Tyrosine Kinase p53/56lyn in Diminished Lipopolysaccharide Priming of Formylmethionylleucyl- phenylalanine-Induced Superoxide Production in Human Newborn Neutrophils

Infection and Immunity ◽

10.1128/iai.72.11.6455-6462.2004 ◽

2004 ◽

Vol 72 (11) ◽

pp. 6455-6462 ◽

Cited By ~ 13

Author(s):

Sen Rong Yan ◽

David M. Byers ◽

Robert Bortolussi

Keyword(s):

Tyrosine Kinase ◽

Tyrosine Kinases ◽

Protein Tyrosine Kinase ◽

Kinase Inhibitor ◽

Critical Role ◽

Polymorphonuclear Neutrophils ◽

Protein Tyrosine Kinases ◽

Protein Tyrosine ◽

Adult Cells

ABSTRACT Human newborns are more susceptible than adults to bacterial infection. With gram-negative bacteria, this may be due to a diminished response of newborn leukocytes to lipopolysaccharide (LPS). Since protein tyrosine kinase inhibition abolishes LPS priming in adult cells, we hypothesized that protein tyrosine kinases may have a critical role in LPS priming of polymorphonuclear neutrophils (PMNs) and that newborn PMNs may have altered protein tyrosine kinase activities. In the present study, we investigated the role of src family protein tyrosine kinases in the LPS response of newborn PMNs compared to adult cells. In a respiratory assay, the LPS-primed increase in formylmethionylleucylphenylalanine (fMLP)-triggered O2 − release by adult PMNs was greatly decreased by PP1 [4-amino-5-(4-methyphenyl)-7-(t-butyl)pyrazolo[3,4-d]pyrimidine], a src kinase inhibitor, to the level of untreated newborn PMNs, in which LPS failed to prime. LPS activated the src-like kinases p59hck (HCK) and p58fgr (FGR) in both adult and newborn PMNs but increased the activation of p53/56 lyn (LYN) only in adult cells. In newborn PMNs, LYN was highly phosphorylated independent of LPS. We evaluated subcellular fractions of PMNs and found that the phosphorylated form of LYN was mainly in the Triton-extractable, cytosolic fraction in adult PMNs, while in newborn cells it was located mainly in Triton-insoluble, granule- and membrane-associated fractions. In contrast, the phosphorylated mitogen-activated protein kinases ERK1/2 and p38 were mainly detected in the cytosol in both adult and newborn PMNs. These data indicate a role for LYN in the regulation of LPS priming. The trapping of phosphorylated LYN in the membrane-granule fraction in newborn PMNs may contribute to the deficiency of newborn cells in responding to LPS stimulation.

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Protein-tyrosine Kinase p72 syk Is Activated by Platelet Activating Factor in Platelets

Thrombosis and Haemostasis ◽

10.1055/s-0038-1648987 ◽

1994 ◽

Vol 72 (06) ◽

pp. 937-941 ◽

Cited By ~ 7

Author(s):

Karim Rezaul ◽

Shigeru Yanagi ◽

Kiyonao Sada ◽

Takanobu Taniguchi ◽

Hirohei Yamamura

Keyword(s):

Tyrosine Kinase ◽

Tyrosine Phosphorylation ◽

Protein Tyrosine Kinase ◽

Kinase Inhibitor ◽

Critical Role ◽

Platelet Activating Factor ◽

Kinase Assay ◽

Potential Candidate ◽

Protein Tyrosine ◽

Induced Aggregation

SummaryIt has been demonstrated that activation of platelets by platelet-activating factor (PAF) results in a dramatic increase in tyrosine phosphorylation of several cellular proteins. We report here that p72 syk is a potential candidate for the protein-tyrosine phosphorylation following PAF stimulation in porcine platelets. Immunoprecipitation kinase assay revealed that PAF stimulation resulted in a rapid activation of p72 syk which peaked at 10 s. The level of activation was found to be dose dependent and could be completely inhibited by the PAF receptor antagonist, CV3988. Phosphorylation at the tyrosine residues of p72 syk coincided with activation of yllsyk. Pretreatment of platelets with aspirin and apyrase did not affect PAF induced activation of p72 syk .Furthermore, genistein, a potent protein-tyrosine-kinase inhibitor, diminished PAF-induced p72 syk activation and Ca2+ mobilization as well as platelet aggregation. These results suggest that p72 syk may play a critical role in PAF-induced aggregation, possibly through regulation of Ca2+ mobilization.

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Prevention of cardiac allograft arteriosclerosis by protein-tyrosine kinase inhibitor selective for platelet-derived growth factor receptor

Transplantation Proceedings ◽

10.1016/s0041-1345(98)01459-6 ◽

1999 ◽

Vol 31 (1-2) ◽

pp. 102 ◽

Cited By ~ 5

Author(s):

P Koskinen ◽

R Sihvola ◽

M Myllärniemi ◽

P Häyry ◽

E Buchdunger ◽

...

Keyword(s):

Growth Factor ◽

Tyrosine Kinase ◽

Tyrosine Kinase Inhibitor ◽

Protein Tyrosine Kinase ◽

Kinase Inhibitor ◽

Growth Factor Receptor ◽

Cardiac Allograft ◽

Platelet Derived Growth Factor ◽

Protein Tyrosine ◽

Protein Tyrosine Kinase Inhibitor

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The role of small molecule Kit protein-tyrosine kinase inhibitors in the treatment of neoplastic disorders

Pharmacological Research ◽

10.1016/j.phrs.2018.04.020 ◽

2018 ◽

Vol 133 ◽

pp. 35-52 ◽

Cited By ~ 28

Author(s):

Robert Roskoski

Keyword(s):

Tyrosine Kinase ◽

Tyrosine Kinase Inhibitors ◽

Small Molecule ◽

Protein Tyrosine Kinase ◽

Kinase Inhibitors ◽

Protein Tyrosine ◽

Neoplastic Disorders

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Lymphocyte-Specific Protein Tyrosine Kinase (LCK) is Involved in the Aryl Hydrocarbon Receptor-Mediated Impairment of Immunoglobulin Secretion in Human Primary B Cells

Toxicological Sciences ◽

10.1093/toxsci/kfy133 ◽

2018 ◽

Vol 165 (2) ◽

pp. 322-334

Author(s):

Jiajun Zhou ◽

Qiang Zhang ◽

Joseph E Henriquez ◽

Robert B Crawford ◽

Norbert E Kaminski

Keyword(s):

B Cells ◽

Tyrosine Kinase ◽

Aryl Hydrocarbon Receptor ◽

Protein Tyrosine Kinase ◽

Specific Protein ◽

Immunoglobulin M ◽

Protein Tyrosine ◽

Immunoglobulin Secretion ◽

Aryl Hydrocarbon

AbstractThe aryl hydrocarbon receptor (AHR) is a cytosolic ligand-activated transcription factor involved in xenobiotic sensing, cell cycle regulation, and cell development. In humans, the activation of AHR by 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD), a high affinity AHR-ligand, impairs the secretion of immunoglobulin M (IgM) to suppress humoral immunity. However, the mechanisms bridging the activation of AHR and the impairment of IgM secretion by human primary B cells remain poorly understood. Recent transcriptomic analysis revealed upregulation of lymphocyte-specific protein tyrosine kinase (LCK) in AHR-activated human primary B cells. LCK is a well-characterized tyrosine kinase that phosphorylates critical signaling proteins involved in activation and cytokine production in T cells. Conversely, the role of LCK in human primary B cells is not well understood. In the current studies, we have verified the transcriptomic finding by detecting AHR-mediated upregulation of LCK protein in human primary B cells. We also confirmed the role of AHR in the upregulation of LCK by using a specific AHR antagonist, which abolished the AHR-mediated increase of LCK. Furthermore, we have confirmed the role of LCK in the AHR-mediated suppression of IgM by using LCK specific inhibitors, which restored the IgM secretion by human B cells in the presence of TCDD. Collectively, the current studies demonstrate a novel role of LCK in IgM response and provide new insights into the mechanism for AHR-mediated impairment of immunoglobulin secretion by human primary B cells.

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