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Inflammation

, Volume 41, Issue 3, pp 984–995 | Cite as

Anti-Inflammatory Effects of Hypophyllanthin and Niranthin Through Downregulation of NF-κB/MAPKs/PI3K-Akt Signaling Pathways

  • Hemavathy Harikrishnan
  • Ibrahim Jantan
  • Md. Areeful Haque
  • Endang Kumolosasi
ORIGINAL ARTICLE

Abstract

Hypophyllanthin (HYP) and niranthin (NIR) are major lignans in Phyllanthus spp. and have been shown to possess strong anti-inflammatory activity. In this study, we investigated the anti-inflammatory effects and the underlying molecular mechanisms of HYP and NIR in in vitro cellular model of LPS-induced U937 macrophages. The effects of HYP and NIR on the production of prostaglandin E2 (PGE2), cyclooxygenase-2 (COX-2), tumor necrosis factor-α (TNF-α), and interleukin-1β (IL-1β) were measured by using ELISA, Western blot, and qRT-PCR. The expressions of signaling molecules related to nuclear factor-kappa B (NF-κB), mitogen-activated protein kinases (MAPKs), and phosphatidylinositol 3′-kinase-Akt (PI3K-Akt) signaling pathways were examined. The role of NF-κB, MAPKs, and Akt signaling pathways was confirmed by using specific inhibitors (BAY 11-7082, U0126, SB202190, SP600125, and LY294002) mediated suppression of TNF-α and COX-2 production. HYP and NIR significantly inhibited the protein and gene levels of COX-2 as well as the downstream signaling products of PGE2, TNF-α, and IL-1β. HYP and NIR also suppressed the inhibitors of kappa B (IκB), IkB kinases (Ikkα/β), NF-κB phosphorylation, and IκB degradation. HYP suppressed the phosphorylation of c-Jun N-terminal kinase (JNK), extracellular signal-regulated kinase (ERK), and p38 while NIR only suppressed JNK and ERK but did not have effect on p38. These results demonstrate that HYP and NIR downregulated COX-2, TNF-α, and IL-1β gene expressions in U937 macrophages by interfering with the activation of NF-κB, MAPKs, and Akt. In conclusion, these lignans have potential to be developed as anti-inflammatory agents targeting the NF-κB, MAPK, and PI3K-Akt pathways.

KEY WORDS

hypophyllanthin niranthin macrophages NF-κB MAPK PI3K-Akt signaling 

Abbreviations

ANOVA

One-way analysis of variance

ATCC

American Type Culture Collection

COX-2

Cyclooxygenase-2

ELISA

Enzyme-linked immunosorbent assay

ERK

Extracellular signal-regulated kinase

FBS

Fetal bovine serum

IL-1β

Interleukin-1 beta

iNOS

Inducible nitric oxide synthase

IκB

I-kappa B kinase

Ikkα/β

IkB kinases

JNK

c-Jun N-terminal kinase

LPS

Lipopolysaccahride

MAPK

Mitogen-activated protein kinase

MyD88

Myeloid differentiation primary response gene 88

NF-κB

Nuclear factor-kappa B

PAMPs

Pathogen-associated molecular patterns

PGE2

Prostaglandin E2

PMA

Phorbol 12-myristate 13-acetate

PI3K-Akt

phosphatidylinositol 3′-kinase-Akt

qRT-PCR

Quantitative real-time polymerase chain reaction

RPMI

Roswell Park Memorial Institute

TBS-T

Tris-buffered saline with Tween 20

TLR4

Toll-like receptor 4

TNF-α

Tumor necrosis factor-alpha

Notes

Acknowledgements

The authors thank the Ministry of Agriculture and Agro-based Industries, Malaysia, for the financial support under the NKEA Research Grant Scheme (NRGS) (Grant No. NH1015D075).

Compliance with Ethical Standards

Conflict of Interest

The authors declare that they have no conflict of interest.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Drug and Herbal Research Center, Faculty of PharmacyUniversiti Kebangsaan MalaysiaKuala LumpurMalaysia

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