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Tetramethylpyrazine Inhibits Platelet Adhesion and Inflammatory Response in Vascular Endothelial Cells by Inhibiting P38 MAPK and NF-κB Signaling Pathways

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Abstract

Damaged vascular endothelial cells after ischemic stroke release inflammatory cytokines and adhesion molecules, which could trigger platelet adhesion to vascular endothelial cells and platelet activation, and accelerate thrombus formation. Tetramethylpyrazine is the main bioactive component of Chuanxiong, which has demonstrated considerable protective effects in cerebrovascular diseases. However, the effect and mechanisms of tetramethylpyrazine on platelet adhesion to ischemia/reperfusion-injured endothelial cells have not been elucidated. In this study, we established an oxygen-glucose deprivation/reoxygenation (OGD/R)–induced brain microvascular endothelial cells (BMECs) injury model to investigate the protective effects of tetramethylpyrazine on platelet adhesion to endothelial cells and potential mechanisms. Experimental results showed that tetramethylpyrazine inhibited platelets adhesion to BMECs, alleviated expression of inflammatory cytokines and adhesion molecules on BMECs, and protected BMECs injured by OGD/R. Furthermore, tetramethylpyrazine could inhibit P38 MAPK and NF-κB activation in injured BMECs by OGD/R and inhibition of P38 MAPK with SB303580 and NF-κB with Bay-11-7082 attenuated the reduction of platelets adhesion to BMECs by tetramethylpyrazine. In conclusion, tetramethylpyrazine protected BMECs and inhibited platelets adhesion to BMECs after OGD/R injury, which was partially mediated by inhibiting P38 MAPK and NF-κB signaling pathways.

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Abbreviations

Akt:

serine/threonine kinase 1

Bax:

BCL2-associated X

Bcl-2:

B cell lymphoma-2

BMECs:

brain microvascular endothelial cells

CCK-8:

Cell Counting Kit-8

CXCR4:

C-X-C motif chemokine receptor 4

CXCR7:

C-X-C motif chemokine receptor 7

DMEM:

Dulbecco’s Modified Eagle’s Medium

EBSS:

Earle’s Balanced Salts

ERK:

extracellular regulated MAP kinase

FBS:

fetal bovine serum

GAPDH:

glyceraldehyde-3-phosphate dehydrogenase

HIF-1α:

hypoxia inducible factor 1 subunit alpha

HMGB1:

high mobility group box 1

HO-1:

heme oxygenase 1

ICAM-1:

intercellular adhesion molecule-1

IL-1β:

interleukin 1 beta

IL-6:

interleukin 6

IL-8:

interleukin 8

LDH:

lactate dehydrogenase

LPS:

lipopolysaccharide

MAPK:

mitogen-activated protein kinase

MCP-1:

macrophage cationic peptide 1

MLKL:

mixed lineage kinase domain-like protein

NF-κB:

nuclear factor-kappa B

Nrf2:

nuclear factor, erythroid 2 like 2

OGD/R:

oxygen-glucose deprivation/reoxygenation

PBS:

phosphate-buffered saline

PGE1:

prostaglandin E1

RIP1:

receptor-interacting protein kinase 1

RIP3:

receptor-interacting protein kinase 3

PIPES:

piperazine-1,4-bis(2-ethanesulfonic acid)

SDF-1:

stromal cell-derived factor 1

TLR4:

toll-like receptor 4

TNF-α:

tumor necrosis factor alpha

VCAM-1:

vascular cell adhesion molecule 1

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Funding

This research was funded by National Natural Science Foundation of China (81622051) and Natural Science Foundation of Tianjin City (15JCYBJC54800).

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    Authors and Affiliations

    1. Tianjin State Key Laboratory of Modern Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin, 300193, China

      Han Zhang, Junhua Zhang & Xiang Fan

    2. Institute of Traditional Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin, 300193, China

      Han Zhang, Weiwei Tang, Shuang Wang, Junhua Zhang & Xiang Fan

    Author notes

    1. Han Zhang and Weiwei Tang contributed equally to this work.

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      1. Han Zhang
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      Contributions

      HZ and XF contributed to the design of the study. HZ, WT, and SW were responsible for the data collection. HZ, WT, and XF analyzed the data. HZ and XF interpreted the data. XF drafted the manuscript. JZ revised the manuscript content. All the authors read and approved the final manuscript.

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      Correspondence to Xiang Fan.

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      Zhang, H., Tang, W., Wang, S. et al. Tetramethylpyrazine Inhibits Platelet Adhesion and Inflammatory Response in Vascular Endothelial Cells by Inhibiting P38 MAPK and NF-κB Signaling Pathways. Inflammation 43, 286–297 (2020). https://doi.org/10.1007/s10753-019-01119-6

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