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Pyrrolidine Dithiocarbamate Activates p38 MAPK and Protects Brain Endothelial Cells From Apoptosis: A Mechanism for the Protective Effect in Stroke?

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Abstract

The antioxidant and inhibitor of nuclear factor κB pyrrolidine dithiocarbamate (PDTC) potently reduces infarct size in various experimental stroke models. In addition, it has been shown to have a favourable safety profile in humans. In this study, we further investigated the mechanistic actions of PDTC on cerebral microvascular endothelial cells as main components of the blood–brain barrier. We propose activation of p38 MAPK by PDTC as an additional protective mechanism. C57/BL6 mice were subjected to transient MCAO for 2 h and treated with PDTC (100 mg/kg) or vehicle i.p. before reperfusion. Infarct size was determined after 24 h. Apoptosis was induced in a cerebral microvascular endothelial cell line and the effect of pretreatment with PDTC and its dependency on p38 MAPK activity was assayed. PDTC administered 2 h after MCAO reduced infarct size by 61% (P < 0.05) and reduces the apoptotic death rate in vivo. In vitro, PDTC reduces the apoptotic death rate of bEnd.3 cells. p38 MAPK was activated by PDTC and its inhibition abrogated the protective effect of PDTC. PDTC reduces infarct size after stroke with a reasonable time window and decreases apoptotic cell death in vivo and in vitro. The attenuation of apoptotic cell death in brain microvascular endothelial cells is dependent on p38 MAPK activity.

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Abbreviations

Bad:

Bcl-2 Antagonist of cell death

Bcl-2:

B-Cell lymphoma 2

CHX:

Cycloheximide

CO:

Carbon monoxide

DETA-NO:

Diethylenetriamine-NO

GSK3:

Glycogen synthase kinase 3

HO-1:

Heme oxygenase-1

IAP:

Inhibitor of apoptosis protein

IκBα:

Inhibitory protein of NF-κB

IKK:

IκB Kinase

JNK:

c-Jun N-terminal kinase

MCAO:

Middle cerebral artery occlusion

MIP-2:

Macrophage inflammatory protein-2

NF-κB:

Nuclear factor κB

p38 MAPK:

p38 Mitogen-activated protein kinase/stress-activated protein kinase

PBS:

Phosphate-buffered solution

PDTC:

Pyrrolidine dithiocarbamate

PI3K:

Phosphatidylinositol 3-kinase

PKB:

Protein kinase B

rt-PA:

Recombinant tissue plasminogen activator

SEM:

Standard error of the mean

STAT:

Signal transducer and activator of transcription

TNFα:

Tumour necrosis factor α

TUNEL:

TdT-Mediated dUTP-biotin nick end labelling

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Acknowledgments

The technical advice of Dr. Peter Lüdike on the MCAO model is gratefully acknowledged. We thank Dr. Alf Theisen and Christa Tandi for providing the infrastructure for the animal studies. W.P. was supported by a grant from the Medical faculty of Johann Wolfgang Goethe University (Patenschaftsstipendium), the Novartis Stiftung für Therapeutische Forschung and by the trust of endowments of Marie Christine Held and Erika Hecker.

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Authors and Affiliations

  1. Klinik für Neurologie, Klinikum der Johann Wolfgang Goethe-Universität, Theodor-Stern-Kai 7, 60590, Frankfurt am Main, Germany

    Waltraud Pfeilschifter, Timo Kahles, Helmuth Steinmetz & Tobias Neumann-Haefelin

  2. Pharmazentrum Frankfurt, Klinikum der Johann Wolfgang Goethe-Universität, Theodor-Stern-Kai 7, 60590, Frankfurt am Main, Germany

    Waltraud Pfeilschifter, Bożena Czech, Britta P. Hoffmann, Marian Sujak & Josef Pfeilschifter

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  1. Waltraud Pfeilschifter
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  2. Bożena Czech
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  3. Britta P. Hoffmann
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  4. Marian Sujak
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  5. Timo Kahles
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  8. Josef Pfeilschifter
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Correspondence to Waltraud Pfeilschifter.

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Waltraud Pfeilschifter and Bozena Czech contributed equally to this work.

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Pfeilschifter, W., Czech, B., Hoffmann, B.P. et al. Pyrrolidine Dithiocarbamate Activates p38 MAPK and Protects Brain Endothelial Cells From Apoptosis: A Mechanism for the Protective Effect in Stroke?. Neurochem Res 35, 1391–1401 (2010). https://doi.org/10.1007/s11064-010-0197-0

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  • DOI: https://doi.org/10.1007/s11064-010-0197-0

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