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Anti-neuroinflammatory effects of tryptanthrin from Polygonum tinctorium Lour. in lipopolysaccharide-stimulated BV2 microglial cells

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Abstract

This study was conducted to isolate the anti-neuroinflammatory component(s) in the 80% EtOH extract of P. tinctoria, and to investigate underlying molecular mechanism of the anti-neuroinflammatory component(s) in LPS-induced BV2 microglial cells. To isolate the active component(s) in the extract, various chromatographic methods were employed, and the structures of the isolated secondary metabolites were determined mainly by analysis of spectroscopic data such as NMR and MS data. Tryptanthrin (1), isolated from P. tinctoria extract, significantly inhibited the protein expression of iNOS and COX-2, and reduced the levels of their products (NO and PGE2) in LPS-stimulated BV2 microglial cells. Tryptanthrin (1) also downregulated the production of pro-inflammatory cytokines such as TNF-α, IL-6, and IL-1β. These anti-neuroinflammatory effects of tryptanthrin (1) was elucidated to be correlated with inactivating NF-κB pathway by interrupting the phosphorylation and degradation of the inhibitor of κB-α protein, and inhibiting the DNA binding activity of NF-κB. In addition, tryptanthrin (1) suppressed the activation of p38 MAPK pathway. Furthermore, tryptanthrin (1) inhibited the TLR4 and MyD88 protein expression in LPS-stimulated BV2 microglial cells. Taken together, it was suggested that tryptanthrin (1) have anti-neuroinflammatory effect by regulating TLR4-MyD88-mediated several inflammatory pathways including p38 and NF-κB pathways in LPS-induced BV2 microglial cells.

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Abbreviations

AD:

Alzheimer’s disease

ALS:

Amyotrophic lateral sclerosis

CC:

Column chromatography

Cdna:

Complementary DNA

CNS:

Central nervous system

COX-2:

Cyclooxygenase-2

DMEM:

Dulbecco’s modified eagle’s medium

DMSO:

Dimethyl sulfoxide

DNA:

Deoxyribonucleic acid

ECL:

Enhanced chemilumescent

ELISA:

Enzyme-linked immunosorbent assay

ERK1/2:

Extracellular signal-regulated kinase

EtOAc:

Ethyl acetate

FBS:

Fetal bovine serum

HCl:

Hydrochloric acid

HPLC:

High-performance liquid chromatography

IL-1β:

Interleukin-1β

IL-6:

Interleukin-6

iNOS:

Inducible nitric oxide synthase

IκB-α:

Inhibitor kappa B-α

JNK:

c-jun N-terminal kinase

LPS:

Lipopolysaccharide

MAPK:

Mitogen-activated protein kinase

MeOH:

Methyl alcohol

mRNA:

messenger RNA

MS:

Mass spectrometry

MTT:

3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide

MyD88:

Myeloid differentiation primary 88

NF-κB:

Nuclear factor-kappa B

NMR:

Nuclear magnetic resonance

NO:

Nitric oxide

PCNA:

Proliferating cell nuclear antigen

PCR:

Polymerase chain reaction

PD:

Parkinson’s disease

PGE2 :

Prostaglandin E2

PMSF:

Phenylmethylsulfonyl fluoride

RNA:

Ribonucleic acid

ROS:

Reactive oxygen species

TLR4:

Toll-like receptor 4

TNF-α:

Tumor necrosis factor-α

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Acknowledgements

This work was supported by the National Research Foundation of Korea (NRF) Grant funded by the Korea Government (NRF-2016R1A2B4007472).

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Author notes

  1. Seungjun Lee and Dong-Cheol Kim have contributed equally to this study.

Authors and Affiliations

  1. Institute of Pharmaceutical Research and Development, College of Pharmacy, Wonkwang University, Iksan, 54538, Republic of Korea

    Seungjun Lee, Dong-Cheol Kim, Youn-Chul Kim & Hyuncheol Oh

  2. Wonpharm Co., LTD, Iksan, 54588, Republic of Korea

    Hum Young Baek

  3. Department of Oriental Medicine Materials, Dongshin University, Naju, 58245, Republic of Korea

    Kyung-Dong Lee

  4. Hanbang Body-Fluid Research Center, Wonkwang University, Iksan, 54538, Republic of Korea

    Youn-Chul Kim & Hyuncheol Oh

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  1. Seungjun Lee
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Correspondence to Hyuncheol Oh.

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Lee, S., Kim, DC., Baek, H.Y. et al. Anti-neuroinflammatory effects of tryptanthrin from Polygonum tinctorium Lour. in lipopolysaccharide-stimulated BV2 microglial cells. Arch. Pharm. Res. 41, 419–430 (2018). https://doi.org/10.1007/s12272-018-1020-8

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