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

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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.

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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

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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).

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Authors and Affiliations

  1. Drug and Herbal Research Center, Faculty of Pharmacy, Universiti Kebangsaan Malaysia, Jalan Raja Muda Abdul Aziz, 50300, Kuala Lumpur, Malaysia

    Hemavathy Harikrishnan, Ibrahim Jantan, Md. Areeful Haque & Endang Kumolosasi

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  1. Hemavathy Harikrishnan
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  2. Ibrahim Jantan
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Correspondence to Ibrahim Jantan.

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Harikrishnan, H., Jantan, I., Haque, M.A. et al. Anti-Inflammatory Effects of Hypophyllanthin and Niranthin Through Downregulation of NF-κB/MAPKs/PI3K-Akt Signaling Pathways. Inflammation 41, 984–995 (2018). https://doi.org/10.1007/s10753-018-0752-4

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