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Falcarindiol inhibits LPS-induced inflammation via attenuating MAPK and JAK-STAT signaling pathways in murine macrophage RAW 264.7 cells

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Abstract

Falcarindiol (FAD) is a natural polyacetylene compound found rich in many plants of the Umbelliferae family. Previously, we isolated FAD from the rhizome of Cnidium officinale Makino, which belongs to the Umbelliferae family and found it to have a significant inhibitory effect on lipopolysaccharide (LPS)-induced production of nitric oxide, a pro-inflammatory molecule in murine macrophage RAW 264.7 cells. In this study, we investigated its effect on the expression of other major pro-inflammatory molecules as well as the mechanism underlying these effects. Pre-treatment of RAW 264.7 cells with FAD suppressed LPS-stimulated mRNA expression of inducible nitric oxide synthase (iNOS), tumor necrosis factor alpha (TNFα), interleukin-6 (IL-6), and interleukin-1 beta (IL-1β) and thereby reduced the respective protein levels. Mechanistic studies demonstrated that FAD attenuated the LPS-induced activation of JNK, ERK, STAT1, and STAT3 signaling molecules. Moreover, we found that FAD did not influence LPS-induced activation of p38 and NFκB signaling pathways. Collectively, this study provides evidence that FAD inhibits the production of major pro-inflammatory molecules in LPS-challenged murine macrophages via suppression of JNK, ERK, and STAT signaling pathways.

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Abbreviations

BCA:

Bicinchoninic acid

BSA:

Bovine serum albumin

cDNA:

Complementary deoxyribonucleic acid

DMEM:

Dulbecco’s modified Eagle’s medium

DMSO:

Dimethyl sulfoxide

E. coli :

Escherichia coli

ERK:

Extracellular signal–regulated kinase

FAD:

Falcarindiol

FBS:

Fetal bovine serum

IL-1β:

Interleukin-1β

HBSS:

Hank’s balanced salt solution

HEPES:

4-(2-Hydroxyethyl)piperazine-1-ethanesulfonic acid, N-(2-hydroxyethyl)piperazine-N′-(2-ethanesulfonic acid)

HRP:

Horseradish peroxidase

IκB:

Inhibitory kappa B protein

IL-6:

Interleukin-6

iNOS:

Inducible nitric oxide synthase

JAK-STAT:

Janus kinase-signal transducers and activators of transcription

JNK:

c-Jun N-terminal kinase

LPS:

Lipopolysaccharide

MAPKs:

Mitogen-activated protein kinases

mRNA:

Messenger RNA

MTT:

(3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide)

NaCl:

Sodium chloride

NFκB:

Nuclear transcription factor kappa B

NO:

Nitric oxide

P38:

P38 Mitogen-activated protein kinase

PBS:

Phosphate-buffered saline

qRT-PCR:

Quantitative real-time polymerase chain reaction

RIPA:

Radioimmunoprecipitation assay

RNA:

Ribonucleic acid

SDS:

Sodium dodecyl sulfate

STAT1:

Signal transducers and activators of transcription factor 1

STAT3:

Signal transducers and activators of transcription factor 3

TLR4:

Toll-like receptor 4

TNF-α:

Tumor necrosis factor α

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Funding

The funding was provided by Forest Science & Technology Projects, Forest Service, Republic of Korea (Grant No. S211316L010110), Kookmin University (KR) research grant (Grant No. 2016)

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Correspondence to Young-Kyoon Kim.

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Venkatesan, T., Choi, YW., Lee, J. et al. Falcarindiol inhibits LPS-induced inflammation via attenuating MAPK and JAK-STAT signaling pathways in murine macrophage RAW 264.7 cells. Mol Cell Biochem 445, 169–178 (2018). https://doi.org/10.1007/s11010-017-3262-z

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  • DOI: https://doi.org/10.1007/s11010-017-3262-z

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