scholarly journals Upregulation of TGF-b-induced HSP27 by HSP90 inhibitors in osteoblasts

2021 ◽  
Author(s):  
Gen Kuroyanagi ◽  
Haruhiko Tokuda ◽  
Kazuhiko Fujita ◽  
Tetsu Kawabata ◽  
Go Sakai ◽  
...  
Keyword(s):  
Protein Kinase ◽  
P38 Mapk ◽  
Transforming Growth Factor ◽  
Bone Cells ◽  
Specific Inhibitor ◽  
Bmp Signaling ◽  
Mitogen Activated Protein Kinase ◽  
Hsp90 Inhibitors ◽  
Jnk Pathway ◽  
Hsp27 Expression

Abstract Background: Heat shock protein (HSP) 90 functions as a molecular chaperone and is constitutively expressed and induced in response to stress in many cell types. We have previously demonstrated that transforming growth factor-b (TGF-b), the most abundant cytokine in bone cells, induces the expression of HSP27 through Smad2, p44/p42 mitogen-activated protein kinase (MAPK), p38 MAPK, and stress-activated protein kinase/c-Jun N-terminal kinase (SAPK/JNK) in mouse osteoblastic MC3T3-E1 cells. This study investigated the effects of HSP90 on the TGF-b-induced HSP27 expression and the underlying mechanism in mouse osteoblastic MC3T3-E1 cells. Methods: Clonal osteoblastic MC3T3-E1 cells were treated with the HSP90 inhibitors and then stimulated with TGF-b. HSP27 expression and the phosphorylation of Smad2, p44/p42 MAPK, p38 MAPK, and SAPK/JNK were evaluated by western blot analysis. Result: HSP90 inhibitors 17-dimethylaminoethylamino-17-demethoxy-geldanamycin (17-DMAG) and onalespib significantly enhanced the TGF-b-induced HSP27 expression. HSP90 inhibitors, geldanamycin, onalespib, and 17-DMAG did not affect the TGF-b-stimulated phosphorylation of Smad2. Geldanamycin did not affect the TGF-b-stimulated phosphorylation of p44/p42 MAPK or p38 MAPK but significantly enhanced the TGF-b-stimulated phosphorylation of SAPK/JNK. Onalespib also increased the TGF-b-stimulated phosphorylation of SAPK/JNK. Furthermore, SP600125, a specific inhibitor for SAPK/JNK, significantly suppressed onalespib or geldanamycin’s enhancing effect of the TGF-b-induced HSP27 expression levels. As for the canonical BMP signaling pathway, BMP-4 failed to induce the expression of HSP27 in osteoblastic MC3T3-E1 cells. Conclusion: Our results strongly suggest that HSP90 inhibitors upregulated the TGF-b-induced HSP27 expression and that these effects of HSP90 inhibitors were mediated through SAPK/JNK pathway in osteoblasts.

scholarly journals The TGFβ-induced phosphorylation and activation of p38 mitogen-activated protein kinase is mediated by MAP3K4 and MAP3K10 but not TAK1

Open Biology ◽  
2013 ◽  
Vol 3 (6) ◽  
pp. 130067 ◽  
Author(s):  
Gopal P. Sapkota
Keyword(s):  
Protein Kinase ◽  
P38 Mapk ◽  
Transforming Growth Factor Beta ◽  
Molecular Mechanisms ◽  
Epithelial To Mesenchymal Transition ◽  
Transforming Growth Factor ◽  
Mitogen Activated Protein Kinase ◽  
Mapk Phosphorylation ◽  
Mesenchymal Transition ◽  
Mitogen Activated Protein

The signalling pathways downstream of the transforming growth factor beta (TGFβ) family of cytokines play critical roles in all aspects of cellular homeostasis. The phosphorylation and activation of p38 mitogen-activated protein kinase (MAPK) has been implicated in TGFβ-induced epithelial-to-mesenchymal transition and apoptosis. The precise molecular mechanisms by which TGFβ cytokines induce the phosphorylation and activation of p38 MAPK are unclear. In this study, I demonstrate that TGFβ-activated kinase 1 (TAK1/MAP3K7) does not play a role in the TGFβ-induced phosphorylation and activation of p38 MAPK in MEFs and HaCaT keratinocytes. Instead, RNAi -mediated depletion of MAP3K4 and MAP3K10 results in the inhibition of the TGFβ-induced p38 MAPK phosphorylation. Furthermore, the depletion of MAP3K10 from cells homozygously knocked-in with a catalytically inactive mutant of MAP3K4 completely abolishes the TGFβ-induced phosphorylation of p38 MAPK, implying that among MAP3Ks, MAP3K4 and MAP3K10 are sufficient for mediating the TGFβ-induced activation of p38 MAPK.

scholarly journals Interleukin-13 Induction of 15-Lipoxygenase Gene Expression Requires p38 Mitogen-Activated Protein Kinase-Mediated Serine 727 Phosphorylation of Stat1 and Stat3

2003 ◽  
Vol 23 (11) ◽  
pp. 3918-3928 ◽  
Author(s):  
Bo Xu ◽  
Ashish Bhattacharjee ◽  
Biswajit Roy ◽  
Hong-Min Xu ◽  
David Anthony ◽  
...  
Keyword(s):  
Protein Kinase ◽  
P38 Mapk ◽  
Kinase Inhibitor ◽  
Specific Inhibitor ◽  
Serine Phosphorylation ◽  
Time Dependent ◽  
Mitogen Activated Protein Kinase ◽  
Human Monocytes ◽  
Interleukin 13 ◽  
Mitogen Activated Protein

ABSTRACT Interleukin-13 (IL-13) is a cytokine secreted by Th2 lymphocytes that is capable of inducing expression of 15-lipoxygenase (15-LO) in primary human monocytes. We recently demonstrated that induction of 15-LO requires the activation of Jak2 and Tyk2 kinases and Stats 1, 3, 5, and 6. Since IL-13-induced 15-LO expression was inhibited by H7 (a serine-threonine kinase inhibitor), we predicted that Stat serine phosphorylation may also be crucial for 15-LO expression. In this study, we present evidence indicating that IL-13-induced 15-LO mRNA expression was detectable as early as 1 h by real-time reverse transcription-PCR. We found that IL-13 induced a time-dependent serine phosphorylation of both Stat1 and Stat3, detectable at 15 min after IL-13 treatment. In addition, the activation of p38 mitogen-activated protein kinase (MAPK) was detected in a time-dependent fashion, with peak phosphorylation at 15 min after IL-13 treatment. SB202190, a p38 MAPK-specific inhibitor, markedly inhibited IL-13-induced Stat1 and Stat3 serine phosphorylation as well as DNA binding. Furthermore, treatment of cells with Stat1 or Stat3 decoys significantly impaired IL-13-induced 15-LO expression. Taken together, our results provide the first evidence that IL-13 induces p38 MAPK phosphorylation/activation, which regulates Stat1 and Stat3 serine 727 phosphorylation. Both of these events are important steps in IL-13-induced 15-LO expression in human monocytes.

Activation of c-Jun N-terminal kinase promotes survival of cardiac myocytes after oxidative stress

2002 ◽  
Vol 362 (3) ◽  
pp. 561-571 ◽  
Author(s):  
Christopher J. DOUGHERTY ◽  
Lori A. KUBASIAK ◽  
Howard PRENTICE ◽  
Peter ANDREKA ◽  
Nanette H. BISHOPRIC ◽  
...  
Keyword(s):  
Cell Fate ◽  
Cardiac Myocytes ◽  
P38 Mapk ◽  
Specific Inhibitor ◽  
Dominant Negative ◽  
Mitogen Activated Protein Kinase ◽  
Jnk Pathway ◽  
Mitogen Activated Protein ◽  
Factor Α ◽  
Jnk Activation

Reperfusion injury occurs when ischaemic tissue is reperfused. It involves the generation and release of reactive oxygen that activates numerous signalling pathways and initiates cell death. Exposure of isolated cardiac myocytes to chronic hypoxia followed by reoxygenation results in the early activation of c-Jun N-terminal kinase (JNK) and death by apoptosis of approx. 30% of the myocytes. Although JNK activation has been described in a number of models of ischaemia/reperfusion, the contribution of JNK activation to cell fate has not been established. Here we report that the activation of JNK by reoxygenation correlates with myocyte survival. Transfection of myocytes with JNK pathway interfering plasmid vectors or infection with adenoviral vectors support the hypothesis that JNK is protective. Transfection or infection with JNK inhibitory mutants increased the rates of apoptosis by almost 2-fold compared with control cultures grown aerobically or subjected to hypoxia and reoxygenation. Caspase 9 activity, measured by LEHD cleavage, increased > 3-fold during reoxygenation and this activity was enhanced significantly at all times in cultures infected with dominant negative JNK adenovirus. Hypoxia—reoxygenation mediated a biphasic (2.6- and 2.9-fold) activation of p38 mitogen-activated protein kinase, as well as a small increase of tumour necrosis factor α (TNFα) secretion, but treatments with the p38 MAPK-specific inhibitor SB203580 or saturating levels of a TNFα-1 blocking antibody provided only partial protection against apoptosis. The results suggest that JNK activation is protective and that the pathway is largely independent of p38 MAPK or secreted TNFα.

p38 Mitogen-activated protein kinase protects glomerular epithelial cells from complement-mediated cell injury

2003 ◽  
Vol 285 (4) ◽  
pp. F765-F774 ◽  
Author(s):  
Lamine Aoudjit ◽  
Monica Stanciu ◽  
Hongping Li ◽  
Serge Lemay ◽  
Tomoko Takano
Keyword(s):  
Protein Kinase ◽  
Epithelial Cells ◽  
P38 Mapk ◽  
Cell Injury ◽  
Transforming Growth Factor ◽  
Cultured Cells ◽  
Mitogen Activated Protein Kinase ◽  
Control Treatment ◽  
Glomerular Epithelial Cells

In the passive Heymann nephritis (PHN) model of rat membranous nephropathy, complement C5b-9 causes sublytic injury of glomerular epithelial cells (GEC). We previously showed that sublytic concentration of C5b-9 triggers a variety of biological events in GEC. In the current study, we demonstrate that complement activates p38 MAPK in GEC and address the role of p38 in complement-mediated cell injury. When cultured rat GEC were stimulated with complement, p38 kinase activity and phosphorylation were increased by ∼2.4-fold, compared with control. Treatment with p38 inhibitors significantly augmented complement-mediated cytotoxicity. In contrast, when the constitutively active mutant of transforming growth factor-β-activated kinase 1 (TAK1), a kinase upstream of p38, was expressed in GEC in an inducible manner, cytotoxicity was significantly reduced, compared with uninduced cells. p38 inhibitors abolished the protective effect of TAK1 expression. By analogy to cultured cells, p38 activity was also increased in glomeruli from rats with PHN and treatment with the p38 inhibitor FR-167653 increased proteinuria. Complement induced phosphorylation of MAPK-associated protein kinase-2 (MAPKAPK-2), a kinase downstream of p38 in GEC. Heat shock protein (HSP27) is a cytoskeleton-interacting substrate of MAPKAPK-2. Overexpression of the wild-type HSP27, but not a non-phosphorylatable mutant, markedly reduced complement-mediated GEC injury. In summary, complement activates p38 MAPK in GEC in vitro and in glomeruli from rats with PHN. The activation of p38 MAPK appears to be cytoprotective for GEC against complement-mediated GEC injury. Phosphorylation of HSP27 may mediate this cytoprotection.

scholarly journals Activation of p38 Mitogen-Activated Protein Kinase Promotes Peritoneal Fibrosis by Regulating Fibrocytes

2012 ◽  
Vol 32 (1) ◽  
pp. 10-19 ◽  
Author(s):  
Satoshi Kokubo ◽  
Norihiko Sakai ◽  
Kengo Furuichi ◽  
Tadashi Toyama ◽  
Shinji Kitajima ◽  
...  
Keyword(s):  
Protein Kinase ◽  
Transforming Growth Factor ◽  
Mapk Pathway ◽  
Abdominal Cavity ◽  
Specific Inhibitor ◽  
Mitogen Activated Protein Kinase ◽  
Type I ◽  
Peritoneal Fibrosis ◽  
Mitogen Activated Protein ◽  
P38mapk Signaling

Background Peritoneal fibrosis is a serious complication of long-term peritoneal dialysis, and yet the precise pathogenic mechanisms of peritoneal fibrosis remain unknown. Fibrocytes participate in tissue fibrosis and express chemokine receptors that are necessary for migration. The p38 mitogen-activated protein kinase (MAPK) pathway regulates the production of chemokines and has been demonstrated to contribute to the pathogenesis of various fibrotic conditions. Accordingly, we used an experimental mouse model of peritoneal fibrosis to examine the dependency of fibrocytes on p38MAPK signaling. Methods Peritoneal fibrosis was induced in mice by the injection of 0.1% chlorhexidine gluconate (CG) into the abdominal cavity. Mice were treated with FR167653, a specific inhibitor of p38MAPK, and immunohistochemical studies were performed to detect fibrocytes and cells positive for phosphorylated p38MAPK. The involvement of p38MAPK in the activation of fibrocytes also was also investigated in vitro. Results Fibrocytes infiltrated peritoneum in response to CG, and that response was accompanied by progressive peritoneal fibrosis. The phosphorylation of p38MAPK, as defined by CD45+ spindle-shaped cells, was detected both in peritoneal mesothelial cells and in fibrocytes. The level of peritoneal expression of CCL2, a chemoattractant for fibrocytes, was upregulated by CG injection, and treatment with FR167653 reduced the number of cells positive for phosphorylated p38MAPK, the peritoneal expression of CCL2, and the extent of peritoneal fibrosis. Pretreatment with FR167653 inhibited the expression of procollagen type I α1 induced by transforming growth factor-β1 Conclusions Our results suggest that p38MAPK signaling contributes to peritoneal fibrosis by regulating fibrocyte function.

An Inhibitor of p38 Mitogen-Activated Protein Kinase Abrogates Procoagulant and Proinflammatory Effects of Antiphospholipid Antibodies In Vivo.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 136-136
Author(s):  
Silvia S. Pierangeli ◽  
Mariano E. Vega-Ostertag ◽  
Xiaowei Liu
Keyword(s):  
Protein Kinase ◽  
Carotid Artery ◽  
P38 Mapk ◽  
Specific Inhibitor ◽  
Mitogen Activated Protein Kinase ◽  
Peritoneal Cells ◽  
Mitogen Activated Protein ◽  
Pre Treatment

Abstract Background: Activation of p38 mitogen-activated protein kinase (p38 MAPK) has been shown to play a fundamental role in antiphospholipid-induced up regulation of tissue factor (TF) expression and function in monocytes and in endothelial cells (ECs) and increased expression of intercellular adhesion molecule -1 (ICAM-1) in vitro. Those effects correlate with the thrombogenic and pro-inflammatory effects of aPL in vivo. However, It is not clear whether aPL-induceTF in vivo. Methods: To examine this question, we treated CD1 male mice, in groups of 4, with IgG from 3 patients with Antiphospholipid Syndrome (IgG-APS) or with control IgG from healthy controls (IgG-NHS), twice. Seventy-two hours after the first injection, the adhesion of leukocytes per capillary venule (#WBC) to EC in cremaster muscle (as an indication of EC activation in vivo), as well the size of an induced thrombus in the femoral vein of the mice were examined. Some mice were infused i.p. with 25 mg/kg of SB203580 (a p38 MAPK-specific inhibitor) 30 minutes prior to the each IgG-APS injection. TF activity was determined using a chromogenic assay that measures the conversion of factor X into Factor Xa, in homogenates of carotid artery, and in peritoneal cells of mice treated with IgG-APS or with IgG-NHS. Expression of TF and ICAM-1 was determined by cyto-ELISA on cultured HUVECs after treatment of the cells with IgG-APS or with IgG-NHS. Results: At the time of the surgical procedures, the mean aCL titer in the sera of the mice injected with IgG-APS was 73 ± 34 GPL. In vivo, IgG-APS increased significantly the #WBC adhering to EC, when compared to control mice (5.25 ± 0.96 vs 1.85 ± 0.72) and these effects were significantly reduced (2.1 ± 0.74), when mice were pre-treated with SB203580. IgG-APS increased significantly the thrombus size when compared to IgG-NHS-treated mice (3189 ± 558 μm2 vs 1468 ± 401 μm2) and SB203580 inhibited this effect by 65%. Treatment of the mice with IgG-APS also induced significantly increased TF function in peritoneal cells and in homogenates of carotid artery when compared to IgG-NHS-treated mice (17.5 ±11.1 pM vs. 0.8 ±0.2 pM and 8.31 ± 1.59 vs 0.69 ± 0.03, respectively). Pre-treatment of the mice with SB203580 abrogated completely those effects (0.61 ± 0.06 pM in peritoneal cells and 0.75 ± 0.28 pM in carotid artery preparations of mice treated with IgG-APS). Significant expression of TF and ICAM-1 was observed in vitro when HUVECs were treated with any of the three IgG-APS. TF upregulation and ICAM-1 expression were significantly reduced by pre-treatment of the cells with SB203580 (49–97% for TF and 25–69% for ICAM-1). Conclusions: The data show that IgG-APS up regulates TF function in vivo, and this correlates with an in vivo pro-inflammatory and pro-thrombotic effect. Importantly, those effects were abrogated in vivo by a p38 MAPK specific inhibitor. These findings may be important in designing new modalities of targeted therapies to treat thrombosis in patients with APS.

scholarly journals The Expression of TGF-b1, p38 MAPK, and ERK-1 Protein in Cleft Affected Tissue of the Lip: An Observational Study

2021 ◽  
Vol 5 (2) ◽  
pp. 82
Author(s):  
Herman Yosef Limpat Wihastyoko ◽  
Erdo Puncak Sidarta
Keyword(s):  
Protein Kinase ◽  
P38 Mapk ◽  
Cleft Lip ◽  
Transforming Growth Factor ◽  
Field Of View ◽  
Mitogen Activated Protein Kinase ◽  
Mitogen Activated Protein ◽  
Affected Tissue ◽  
The Mean ◽  
Tgf Β1

Background: Cleft lip is a congenital birth defect caused by many proteins. Transforming growth factor (TGF)-β1, p38 mitogen-activated protein kinase (MAPK) and extracellular signal-regulated protein kinase (ERK)-1 are proteins which regulate proliferation and apoptosis role during intrauterine period. This study aimed to observe the expression of these proteins in cleft affected tissue of the lip.Materials and Methods: A descriptive study by examining the TGF-β1, p38 MAPK, and ERK-1 immunohistochemical expression of cleft affected tissue of the lip was conducted. Subjects were patients that were participating for the social event held by Plastic Surgery Department, Faculty of Medicine, Univesitas Brawijaya, on December 3-12, 2012 in Nusa Tenggara Timur. Excess lip mucosa (waste tissue) during the operation were stored in 10% formalin then stained by immunohistochemistry for TGF-β1, p38 MAPK, and ERK-1. We counted the average protein expression under the light microscope with 1000x magnification for 20 different fields of view, randomly.Results: Paraffin blocks from 30 subjects were selected. The mean p38 MAPK expression was found to be highest, with the average of 8 per field of view; followed by the mean TGF-β1 expression, with the average of 5 per field of view; and the mean ERK-1 expression was found to be the lowest, with the average of 2 per field of view. Conclusion: Expression of p38 MAPK and TGF-β1 are higher than ERK-1, suggesting that p38 MAPK is in the same signalling pathway as TGF-β1, while ERK-1 is lower, as its role as anti-apoptotic. This is consistent with several previous studies showing that all proteins took part in the development of cleft lip or craniofacial development. Further study needs to be conducted to determine which protein plays the bigger role.Keywords: cleft lip, TGF-β1, p38 MAPK, ERK-1

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