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Schisandrin B elicits a glutathione antioxidant response and protects against apoptosis via the redox-sensitive ERK/Nrf2 pathway in H9c2 cells

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Abstract

This study investigated the signal transduction pathway involved in the cytoprotective action of (−)schisandrin B [(−)Sch B, a stereoisomer of Sch B]. Using H9c2 cells, the authors examined the effects of (−)Sch B on MAPK and Nrf2 activation, as well as the subsequent eliciting of glutathione response and protection against apoptosis. Pharmacological tools, such as cytochrome P-450 (CYP) inhibitor, antioxidant, MAPK inhibitor, and Nrf2 RNAi, were used to delineate the signaling pathway. (−)Sch B caused a time-dependent activation of MAPK in H9c2 cells, with the degree of ERK activation being much larger than that of p38 or JNK. The MAPK activation was followed by an increase in the level of nuclear Nrf2, an indirect measure of Nrf2 activation, and the eliciting of a glutathione antioxidant response. The activation of MAPK and Nrf2 seemed to involve oxidants generated from a CYP-catalyzed reaction with (−)Sch B. Both ERK inhibition by U0126 and Nrf2 suppression by Nrf2 RNAi transfection largely abolished the cytoprotection against hypoxia/reoxygenation-induced apoptosis in (−)Sch B-pretreated cells. (−)Sch B pretreatment potentiated the reoxygenation-induced ERK activation, whereas both p38 and JNK activations were suppressed. Under the condition of ERK inhibition, Sch B treatment did not protect against ischemia/reperfusion injury in an ex vivo rat heart model. The results indicate that (−)Sch B triggers a redox-sensitive ERK/Nrf2 signaling, which then elicits a cellular glutathione antioxidant response and protects against hypoxia/reoxygenation-induced apoptosis in H9c2 cells. The ERK-mediated signaling is also likely involved in the cardioprotection afforded by Sch B in vivo.

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Abbreviations

ABT:

1-Amino-benzotriazole

AIF:

Apoptosis inducing factor

CYP:

Cytochrome P-450

DMTU:

Dimethylthiouracil

DTT:

Dithiothreitol

EpRE:

Electrophile response element

ERK:

Extracellular signal-regulated protein kinase

GCL:

γ-Glutamyl cysteine ligase

G6PDH:

Glucose-6-phosphate dehydrogenase

GR:

Glutathione reductase

GSH:

Reduced glutathione

JNK:

C-jun-NH2-terminal kinases

LDH:

Lactate dehydrogenase

MAPK:

Mitogen-activated protein kinases

mGCL:

Modulatory subunit of GCL

Nrf2:

Nuclear factor erythroid 2-related factor 2

p38:

p38 MAPK

PMSF:

Phenylmethylsulphonyl fluoride

ROS:

Reactive oxygen species

Sch B:

Schisandrin B

SDS:

Sodium docecyl sulfate

Trx:

Thioredoxin-1

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Acknowledgments

This study was supported by a GRF Grant (Project number: 661107) (Principal Investigator Dr. K.M. Ko) from the Research Grants Council, Hong Kong.

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  1. Section of Biochemistry and Cell Biology, Division of Life Science, The Hong Kong University of Science and Technology, Clear Water Bay, Hong Kong SAR, China

    Po Yee Chiu, Na Chen, Po Kuan Leong, Hoi Yan Leung & Kam Ming Ko

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  1. Po Yee Chiu
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Correspondence to Kam Ming Ko.

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Chiu, P.Y., Chen, N., Leong, P.K. et al. Schisandrin B elicits a glutathione antioxidant response and protects against apoptosis via the redox-sensitive ERK/Nrf2 pathway in H9c2 cells. Mol Cell Biochem 350, 237–250 (2011). https://doi.org/10.1007/s11010-010-0703-3

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