Methotrexate promotes platelet apoptosis via JNK-mediated mitochondrial damage: Alleviation by N-acetylcysteine and N-acetylcysteine amide

Manoj Paul; Mahadevappa Hemshekhar; Ram M. Thushara; Mahalingam S. Sundaram; Somanathapura K. NaveenKumar; Shivanna Naveen; Sannaningaiah Devaraja; Kumar Somyajit; Robert West; Basappa; Siddaiah C. Nayaka; Uzma I. Zakai; Ganesh Nagaraju; Kanchugarakoppal S. Rangappa; Kempaiah Kemparaju; Kesturu S.Girish Girish

doi:10.1371/journal.pone.0127558

Methotrexate promotes platelet apoptosis via JNK-mediated mitochondrial damage: Alleviation by N-acetylcysteine and N-acetylcysteine amide

Manoj Paul
, Mahadevappa Hemshekhar
, Ram M. Thushara
, Mahalingam S. Sundaram
, Somanathapura K. NaveenKumar
, Shivanna Naveen
, Sannaningaiah Devaraja
, Kumar Somyajit
, Robert West
, Basappa
, Siddaiah C. Nayaka
, Uzma I. Zakai
, Ganesh Nagaraju
, Kanchugarakoppal S. Rangappa
, Kempaiah Kemparaju
, Kesturu S.Girish Girish

University of Mysore
Manitoba Centre for Proteomics and Systems Biology
Defence Food Research Laboratory India
Tumkur University
University of Wisconsin–Madison
Bangalore University
Indian Institute of Science

Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › peer review

Abstract

Thrombocytopenia in methotrexate (MTX)-treated cancer and rheumatoid arthritis (RA) patients connotes the interference of MTX with platelets. Hence, it seemed appealing to appraise the effect of MTX on platelets. Thereby, the mechanism of action of MTX on platelets was dissected. MTX (10 μM) induced activation of pro-apoptotic proteins Bid, Bax and Bad through JNK phosphorylation leading to Δψm dissipation, cytochrome c release and caspase activation, culminating in apoptosis. The use of specific inhibitor for JNK abrogates the MTX-induced activation of pro-apoptotic proteins and downstream events confirming JNK phosphorylation by MTX as a key event. We also demonstrate that platelet mitochondria as prime sources of ROS which plays a central role in MTX-induced apoptosis. Further, MTX induces oxidative stress by altering the levels of ROS and glutathione cycle. In parallel, the clinically approved thiol antioxidant N-acetylcysteine (NAC) and its derivative N-acetylcysteine amide (NACA) proficiently alleviate MTX-induced platelet apoptosis and oxidative damage. These findings underpin the dearth of research on interference of therapeutic drugs with platelets, despite their importance in human health and disease. Therefore, the use of antioxidants as supplementary therapy seems to be a safe bet in pathologies associated with altered platelet functions.

Originalsprog	Engelsk
Artikelnummer	e0127558
Tidsskrift	PLOS ONE
Vol/bind	10
Udgave nummer	6
Antal sider	15
ISSN	1932-6203
DOI	https://doi.org/10.1371/journal.pone.0127558
Status	Udgivet - jun. 2015
Udgivet eksternt	Ja

Bibliografisk note

Funding Information:
Manoj Paul and Mahalingam S. Sundaram thank UGC-BSR for the research fellowship. Authors thank Pavan MS, Sharath BN, IISc Bangalore and Mr. Jagadish S, UOM Mysore for their kind help during the study and also thank Vashishta from the IISc FACS facility, Bangalore. Authors also thank Central instrumentation facility, Institute of excellence (IOE), University of Mysore, Mysore.

Publisher Copyright:
© 2015 Paul et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Finansiering

Manoj Paul and Mahalingam S. Sundaram thank UGC-BSR for the research fellowship. Authors thank Pavan MS, Sharath BN, IISc Bangalore and Mr. Jagadish S, UOM Mysore for their kind help during the study and also thank Vashishta from the IISc FACS facility, Bangalore. Authors also thank Central instrumentation facility, Institute of excellence (IOE), University of Mysore, Mysore.

Dokumenter og links

10.1371/journal.pone.0127558Licens: CC BY

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Citationsformater

Paul, M., Hemshekhar, M., Thushara, R. M., Sundaram, M. S., NaveenKumar, S. K., Naveen, S., Devaraja, S., Somyajit, K., West, R., Basappa, Nayaka, S. C., Zakai, U. I., Nagaraju, G., Rangappa, K. S., Kemparaju, K., & Girish, K. S. G. (2015). Methotrexate promotes platelet apoptosis via JNK-mediated mitochondrial damage: Alleviation by N-acetylcysteine and N-acetylcysteine amide. PLOS ONE, 10(6), Artikel e0127558. https://doi.org/10.1371/journal.pone.0127558

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title = "Methotrexate promotes platelet apoptosis via JNK-mediated mitochondrial damage: Alleviation by N-acetylcysteine and N-acetylcysteine amide",

abstract = "Thrombocytopenia in methotrexate (MTX)-treated cancer and rheumatoid arthritis (RA) patients connotes the interference of MTX with platelets. Hence, it seemed appealing to appraise the effect of MTX on platelets. Thereby, the mechanism of action of MTX on platelets was dissected. MTX (10 μM) induced activation of pro-apoptotic proteins Bid, Bax and Bad through JNK phosphorylation leading to Δψm dissipation, cytochrome c release and caspase activation, culminating in apoptosis. The use of specific inhibitor for JNK abrogates the MTX-induced activation of pro-apoptotic proteins and downstream events confirming JNK phosphorylation by MTX as a key event. We also demonstrate that platelet mitochondria as prime sources of ROS which plays a central role in MTX-induced apoptosis. Further, MTX induces oxidative stress by altering the levels of ROS and glutathione cycle. In parallel, the clinically approved thiol antioxidant N-acetylcysteine (NAC) and its derivative N-acetylcysteine amide (NACA) proficiently alleviate MTX-induced platelet apoptosis and oxidative damage. These findings underpin the dearth of research on interference of therapeutic drugs with platelets, despite their importance in human health and disease. Therefore, the use of antioxidants as supplementary therapy seems to be a safe bet in pathologies associated with altered platelet functions.",

author = "Manoj Paul and Mahadevappa Hemshekhar and Thushara, \{Ram M.\} and Sundaram, \{Mahalingam S.\} and NaveenKumar, \{Somanathapura K.\} and Shivanna Naveen and Sannaningaiah Devaraja and Kumar Somyajit and Robert West and Basappa and Nayaka, \{Siddaiah C.\} and Zakai, \{Uzma I.\} and Ganesh Nagaraju and Rangappa, \{Kanchugarakoppal S.\} and Kempaiah Kemparaju and Girish, \{Kesturu S.Girish\}",

note = "Publisher Copyright: {\textcopyright} 2015 Paul et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.",

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Paul, M, Hemshekhar, M, Thushara, RM, Sundaram, MS, NaveenKumar, SK, Naveen, S, Devaraja, S, Somyajit, K, West, R, Basappa, Nayaka, SC, Zakai, UI, Nagaraju, G, Rangappa, KS, Kemparaju, K & Girish, KSG 2015, 'Methotrexate promotes platelet apoptosis via JNK-mediated mitochondrial damage: Alleviation by N-acetylcysteine and N-acetylcysteine amide', PLOS ONE, bind 10, nr. 6, e0127558. https://doi.org/10.1371/journal.pone.0127558

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T1 - Methotrexate promotes platelet apoptosis via JNK-mediated mitochondrial damage

T2 - Alleviation by N-acetylcysteine and N-acetylcysteine amide

AU - Paul, Manoj

AU - Hemshekhar, Mahadevappa

AU - Thushara, Ram M.

AU - Sundaram, Mahalingam S.

AU - NaveenKumar, Somanathapura K.

AU - Naveen, Shivanna

AU - Devaraja, Sannaningaiah

AU - Somyajit, Kumar

AU - West, Robert

AU - Basappa,

AU - Nayaka, Siddaiah C.

AU - Zakai, Uzma I.

AU - Nagaraju, Ganesh

AU - Rangappa, Kanchugarakoppal S.

AU - Kemparaju, Kempaiah

AU - Girish, Kesturu S.Girish

N1 - Publisher Copyright: © 2015 Paul et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

PY - 2015/6

Y1 - 2015/6

N2 - Thrombocytopenia in methotrexate (MTX)-treated cancer and rheumatoid arthritis (RA) patients connotes the interference of MTX with platelets. Hence, it seemed appealing to appraise the effect of MTX on platelets. Thereby, the mechanism of action of MTX on platelets was dissected. MTX (10 μM) induced activation of pro-apoptotic proteins Bid, Bax and Bad through JNK phosphorylation leading to Δψm dissipation, cytochrome c release and caspase activation, culminating in apoptosis. The use of specific inhibitor for JNK abrogates the MTX-induced activation of pro-apoptotic proteins and downstream events confirming JNK phosphorylation by MTX as a key event. We also demonstrate that platelet mitochondria as prime sources of ROS which plays a central role in MTX-induced apoptosis. Further, MTX induces oxidative stress by altering the levels of ROS and glutathione cycle. In parallel, the clinically approved thiol antioxidant N-acetylcysteine (NAC) and its derivative N-acetylcysteine amide (NACA) proficiently alleviate MTX-induced platelet apoptosis and oxidative damage. These findings underpin the dearth of research on interference of therapeutic drugs with platelets, despite their importance in human health and disease. Therefore, the use of antioxidants as supplementary therapy seems to be a safe bet in pathologies associated with altered platelet functions.

AB - Thrombocytopenia in methotrexate (MTX)-treated cancer and rheumatoid arthritis (RA) patients connotes the interference of MTX with platelets. Hence, it seemed appealing to appraise the effect of MTX on platelets. Thereby, the mechanism of action of MTX on platelets was dissected. MTX (10 μM) induced activation of pro-apoptotic proteins Bid, Bax and Bad through JNK phosphorylation leading to Δψm dissipation, cytochrome c release and caspase activation, culminating in apoptosis. The use of specific inhibitor for JNK abrogates the MTX-induced activation of pro-apoptotic proteins and downstream events confirming JNK phosphorylation by MTX as a key event. We also demonstrate that platelet mitochondria as prime sources of ROS which plays a central role in MTX-induced apoptosis. Further, MTX induces oxidative stress by altering the levels of ROS and glutathione cycle. In parallel, the clinically approved thiol antioxidant N-acetylcysteine (NAC) and its derivative N-acetylcysteine amide (NACA) proficiently alleviate MTX-induced platelet apoptosis and oxidative damage. These findings underpin the dearth of research on interference of therapeutic drugs with platelets, despite their importance in human health and disease. Therefore, the use of antioxidants as supplementary therapy seems to be a safe bet in pathologies associated with altered platelet functions.

U2 - 10.1371/journal.pone.0127558

DO - 10.1371/journal.pone.0127558

M3 - Journal article

C2 - 26083398

AN - SCOPUS:84939209556

SN - 1932-6203

VL - 10

JO - PLOS ONE

JF - PLOS ONE

IS - 6

M1 - e0127558

ER -

Methotrexate promotes platelet apoptosis via JNK-mediated mitochondrial damage: Alleviation by N-acetylcysteine and N-acetylcysteine amide

Abstract

Bibliografisk note

Finansiering

Dokumenter og links

SDU link

Fingeraftryk

Citationsformater