Chronic ethanol feeding potentiates Fas Jo2-induced hepatotoxicity: Role of CYP2E1 and TNF-α and activation of JNK and P38 MAP kinase

Abstract Cytokines such as TNF α, IFN γ and others have been implicated in the pathogenesis of ineffective hematopoiesis in MDS and are thought to lead to the high rate of apoptosis in hematopoietic progenitors. The p38 Mitogen Activated Protein Kinase (MAPK) is an evolutionary conserved enzyme that is involved in many cellular processes including stress signaling. We have previously shown that the p38 MAP kinase is strongly activated by IFNs, TNF α, TGF β and other inhibitory cytokines in normal primary hematopoietic progenitors and plays an important role in the negative regulation of normal hematopoiesis. In the present study, we determined the role of the p38 MAPK in the pathogenesis of MDS evaluated its inhibition as a potential therapeutic strategy in this disease. p38 MAPK inhibition was achieved by the use of a novel p38 inhibitor - SD-282, a specific inhibitor of p38α MAP kinase. SD-282 performs very similarly in animal and cell models to a p38 inhibitor now in the clinic. We also transfected primary hematopoietic cells with flurescent labeled siRNAs against p38 and successfully downregulated the levels of the protein. Using these approaches, we demonstrate that pharmacological inhibition of the p38 MAPK can reverse the growth inhibitory effects of TNF α and IFN γ on erythroid and myeloid colony formation. This reversal of TNF α mediated inhibition correlates with significant reduction of apoptosis seen in human hematopoeitic progenitors pretreated with p38 inhibitor SD-282. Having established the importance of p38 MAPK in cytokine mediated inhibition of normal hematopoiesis, we performed colony forming assays with bone marrow CD34+ cells from 8 patients with MDS in the presence of either pharmacologic or siRNA based inhibitors of p38. All patients had refractory cytopenias with multilineage dysplasia. Our data indicates that SD-282 treatment strongly enhances both erythroid and myeloid colony formation in MDS CD34+ bone marrow cells in vitro. This increase was not observed when these progenitors were grown in the presence of negative controls - SB 202474 and the MEK inhibitor PD 98059. Similarly, an increase in hematopoietic colony formation, though of a lesser magnitude was seen when MDS bone marrow progenitors were transfected with siRNAs against p38 MAPK. To further determine the role of cytokines in the pathogenesis of MDS, we also used bone marrow derived sera from the same MDS patients. Our studies show exposure to patient derived sera led to the phosphorylation/activation of p38 MAPK in normal hematopoietic progenitors when compared to sera from healthy volunteers. Our studies also demonstrate that bone marrow derived sera from MDS patients can inhibit erythroid and myeloid colony formation of normal hematopoietic progenitors. This inhibition can be reversed by blocking p38 MAPK using SD-282, other p38 inhibitors and siRNAs. This finding confirms the role of marrow cytokine /serum factors in the ineffective hematopoiesis seen in MDS and suggests the importance of p38 MAPK activation in this phenomenon. Thus our studies show the p38 MAPK may be a common effector of inhibitory cytokine signaling in normal and MDS hematopoietic cells. These results provide a strong rationale for using p38 inhibition as a novel treatment strategy for MDS. Supported by Harris Methodist Foundation Grant, VISN-17 New Investigator Grant and VA Research Corp Grant to AV.

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Barrier Dysfunction of the Corneal Endothelium in Response to TNF-α: Role of p38 MAP Kinase

Investigative Opthalmology & Visual Science ◽

10.1167/iovs.09-4343 ◽

2010 ◽

Vol 51 (3) ◽

pp. 1575 ◽

Cited By ~ 34

Author(s):

Mahesh Shivanna ◽

Gangaraju Rajashekhar ◽

Sangly P. Srinivas

Keyword(s):

Map Kinase ◽

Corneal Endothelium ◽

P38 Map Kinase ◽

Barrier Dysfunction ◽

Tnf Α

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Tumour necrosis factor-α-induced phosphorylation and activation of cytosolic phospholipase A2 are abrogated by an inhibitor of the p38 mitogen-activated protein kinase cascade in human neutrophils

Biochemical Journal ◽

10.1042/bj3190017 ◽

1996 ◽

Vol 319 (1) ◽

pp. 17-20 ◽

Cited By ~ 91

Author(s):

Waltraut H WATERMAN ◽

Thaddeus F. P. MOLSKI ◽

Chi-Kuang HUANG ◽

Jerry L. ADAMS ◽

Ramadan I. SHA'AFI

Keyword(s):

Map Kinase ◽

P38 Map Kinase ◽

Mitogen Activated Protein Kinase ◽

Human Neutrophils ◽

Tnf Α ◽

Mitogen Activated Protein ◽

Kinase Cascade ◽

Factor Α ◽

Sb 203580

The role of the newly identified p38 mitogen-activated protein kinase (MAP kinase) in terminally differentiated cells, such as human neutrophils, is totally unknown. In order to examine the possible role of this MAP kinase in the phosphorylation and activation of cytoplasmic phospholipase A2 (cPLA2), we tested the effect of the recently synthesized inhibitor of p38 MAP kinase, SB 203580, on the phosphorylation and activation of both p38 MAP kinase and cPLA2. We found that while tumour necrosis factor-α (TNF-α)-stimulated tyrosine phosphorylation of p38 MAP kinase is affected only slightly by SB 203580, its stimulated kinase activity is greatly reduced in human neutrophils in suspension treated with this inhibitor. Furthermore, the TNF-α-stimulated phosphorylation and activation of cPLA2 are completely abolished in cells treated with SB 203580. Based on these data, it is reasonable to conclude that an SB 203580-sensitive kinase, or kinases and/or phosphatases, are involved in the phosphorylation and activation of cPLA2 in intact human neutrophils in suspension stimulated by TNF-α. The possible role of the p38 MAP kinase cascade in the phosphorylation and activation of cPLA2 is discussed.

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HIV-1 Tat protein induces IL-10 production by an alternative TNF-α-independent pathway in monocytes: Role of PKC-δ and p38 MAP kinase

Cellular Immunology ◽

10.1016/j.cellimm.2008.04.015 ◽

2008 ◽

Vol 253 (1-2) ◽

pp. 45-53 ◽

Cited By ~ 25

Author(s):

Kaoutar Leghmari ◽

Yamina Bennasser ◽

Jean Tkaczuk ◽

Elmostafa Bahraoui

Keyword(s):

Map Kinase ◽

P38 Map Kinase ◽

Tat Protein ◽

Tnf Α ◽

Hiv 1

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677 Inhibition of p38 MAP kinase improves endothelial dysfunction in chronic heart failure: role of vascular superoxide anion production

European Journal of Heart Failure Supplements ◽

10.1016/s1567-4215(03)90422-x ◽

2003 ◽

Vol 2 (1) ◽

pp. 139

Author(s):

J WIDDER ◽

T BEHR ◽

K HU ◽

M BEROVA ◽

C ANGERMANN ◽

...

Keyword(s):

Heart Failure ◽

Chronic Heart Failure ◽

Endothelial Dysfunction ◽

Map Kinase ◽

Superoxide Anion ◽

P38 Map Kinase ◽

Superoxide Anion Production ◽

Anion Production

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Species Comparison of the Role of p38 MAP Kinase in the Female Reproductive System

Journal of Toxicologic Pathology ◽

10.1293/tox.22.109 ◽

2009 ◽

Vol 22 (2) ◽

pp. 109-124 ◽

Cited By ~ 5

Author(s):

Zaher A. Radi ◽

Rosemary A. Marusak ◽

Dale L. Morris

Keyword(s):

Map Kinase ◽

Reproductive System ◽

P38 Map Kinase ◽

Female Reproductive System ◽

Species Comparison

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TNF-α and serum induce SKALP/elafin gene expression in human keratinocytes by a p38 MAP kinase-dependent pathway

Archives of Dermatological Research ◽

10.1007/s004030050475 ◽

2000 ◽

Vol 292 (4) ◽

pp. 180-187 ◽

Cited By ~ 68

Author(s):

R. Pfundt ◽

M. Wingens ◽

M. Bergers ◽

M. Zweers ◽

M. Frenken ◽

...

Keyword(s):

Gene Expression ◽

Map Kinase ◽

Human Keratinocytes ◽

P38 Map Kinase ◽

Tnf Α ◽

Dependent Pathway

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Aqueous extract of Magnolia officinalis mediates proliferative capacity, p21WAF1 expression and TNF-α-induced NF-κB activity in human urinary bladder cancer 5637 cells; involvement of p38 MAP kinase

Oncology Reports ◽

10.3892/or.18.3.729 ◽

2007 ◽

Author(s):

Se-Jung Lee ◽

Hong-Man Kim ◽

Young-Hwa Cho ◽

Keerang Park ◽

Eun-Jung Kim ◽

...

Keyword(s):

Bladder Cancer ◽

Urinary Bladder ◽

Map Kinase ◽

Aqueous Extract ◽

Urinary Bladder Cancer ◽

P38 Map Kinase ◽

Proliferative Capacity ◽

Human Urinary Bladder ◽

Tnf Α ◽

Magnolia Officinalis

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Differential role of cAMP, cGMP and Ca2+ and involvement of kinases and CBP-CREB-CRE pathway in regulation of arylalkylamine N-acetyltransferase 2 gene in the pineal organ of air-breathing catfish, Clarias gariepinus

10.21203/rs.3.rs-422474/v1 ◽

2021 ◽

Author(s):

Hijam Nonibala ◽

Braj Bansh Prasad Gupta

Keyword(s):

Gene Expression ◽

Map Kinase ◽

Gene Transcription ◽

Pineal Organ ◽

Clarias Gariepinus ◽

Specific Inhibitor ◽

P38 Map Kinase ◽

Camp And Cgmp

Abstract Transcription of arylalkylamine N-acetyltransferase 2 (aanat2) gene leads to formation of AANAT2 - the rate-limiting enzyme in melatonin synthesis pathway in photosensitive fish pineal organ. However, unlike in avian and mammalian pineal gland, there is practically no information on signal transduction pathway(s) involved in regulation of aanat2 gene transcription in the fish pineal organ. Therefore, we investigated the role of important molecular components of signalling via cAMP, cGMP, Ca2+ involving PKA, PKG, PKC, MeK and p38 MAP kinase as well as possible role of serine/threonine phosphatases, CREB and CBP using their specific inhibitors and/or activators in aanat2 gene transcription in the fish pineal organ maintained under in vitro culture-conditions. db-cAMP and db-cGMP stimulated the expression of aanat2 gene. db-cAMP- and cGMP-induced aanat2 gene expression was significantly reduced in the presence of H-89 (specific inhibitor of PKA), KT5823 (specific inhibitor of PKG), chelerythrine chloride (specific inhibitor of PKC), U0126 ethanolate (specific inhibitor of MeK) and SB 202190 monohydrochloride hydrate (specific inhibitor of p38 MAP kinase). Inhibitors of PP1 and PP2A significantly increased aanat2 gene expression as well as significantly reduced cAMP- and cGMP-induced gene transcription, while inhibitor of PP2B had no effect on aanat2 gene expression. Inhibitors of both CREB and CBP-CREB interaction completely blocked cAMP-induced aanat2 gene transcription. Based on these findings, we suggest that cAMP, cGMP and Ca2+ stimulate aanat2 gene transcription via PKA, PKG and PKC, respectively. Further, protein phosphatases and CBP-CREB-CRE pathway are actively involved in regulation of on aanat2 gene expression in the fish pineal organ.

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