Dataset Information

Siegesbeckia pubescens Makino inhibits Pam₃CSK₄-induced inflammation in RAW 264.7 macrophages through suppressing TLR1/TLR2-mediated NF-?B activation.

ABSTRACT:

Background

Siegesbeckia pubescens Makino (SP) is one of the important plant origins for the anti-inflammatory Chinese herbal medicine of Siegesbeckiae Herba. The current investigations indicated that the anti-inflammatory effects of SP were associated with the toll-like receptors (TLRs)-mediated nuclear factor-?B (NF-?B) and the mitogen-activated protein kinase (MAPK) signaling pathways.

Methods

Raw 264.7 macrophages were pretreated with the 50% ethanol extract of SP (SPE, 50-200 µg/mL) and then co-treated with Pam₃CSK₄ (200 ng/mL) for another 12 h. The inhibitory effect of SPE on Pam₃CSK₄-stimulated NO release and post-inflammatory cytokines secretions were determined using Griess reagent and Elisa kits, respectively. The influence of SPE on NF-?B and MAPKs signaling relevant proteins was measured by Western blotting analysis, while the intracellular nitric oxide (NO) generation and NF-?B/p65 nuclear translocation were determined using Leica TCS SP8 laser scanning confocal microscope. Moreover, the effect of SPE on luciferase reporter gene in NF-?B-luc DNA transfected raw 264.7 cells was determined using the Dual-Glo luciferase assay system kit.

Results

SPE dose-dependently (50-200 µg/mL) attenuated Pam₃CSK₄-induced NO release, post-inflammatory cytokines (IL-6, TNF-? and MCP-1) secretions and intracellular NO generation in raw 264.7 cells. Biologically, SPE suppressed Pam₃CSK₄-induced expressions of cyclooxygenase-2 (COX-2), inducible nitric oxide synthase (iNOS), phosphorylation of NF-?B/p65 and I?B?, but did not significantly show effect on the proteins involved in MAPKs signaling (p38, ERK and JNK). The results were further confirmed by NF-?B-luc reporter gene assay and p65 nuclear translocation assay.

Conclusions

In conclusion, SPE ameliorated Pam₃CSK₄-induced inflammation in raw 264.7 cells through suppressing TLR 1/2-mediated NF-?B activation.

SUBMITTER: Sang W

PROVIDER: S-EPMC6034227 | biostudies-literature | 2018

REPOSITORIES: biostudies-literature

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Publications

<i>Siegesbeckia pubescens</i> Makino inhibits Pam<sub>3</sub>CSK<sub>4</sub>-induced inflammation in RAW 264.7 macrophages through suppressing TLR1/TLR2-mediated NF-κB activation.

Sang Wei W Zhong Zhangfeng Z Linghu Kegang K Xiong Wei W Tse Anfernee Kai Wing AKW Cheang Wai San WS Yu Hua H Wang Yitao Y

Chinese medicine 20180705

<h4>Background</h4><i>Siegesbeckia pubescens</i> Makino (SP) is one of the important plant origins for the anti-inflammatory Chinese herbal medicine of Siegesbeckiae Herba. The current investigations indicated that the anti-inflammatory effects of SP were associated with the toll-like receptors (TLRs)-mediated nuclear factor-κB (NF-κB) and the mitogen-activated protein kinase (MAPK) signaling pathways.<h4>Methods</h4>Raw 264.7 macrophages were pretreated with the 50% ethanol extract of SP (SPE, ...[more]

PMID: 30002726

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Project description:Mycobacterium ulcerans produces a macrolide exotoxin, mycolactone which suppresses immune cells activity, is toxic to most cells and the key virulence factor in the pathogenesis of Buruli ulcer disease. Mycolactone is reported to mediate the production of reactive oxygen species in keratinocytes; cells that play critical role in wound healing. Increased levels of reactive oxygen species have been shown to disrupt the well-ordered process of wound repair; hence, the function of wound-healing cells such as macrophages, keratinocytes, and fibroblast could be impaired in the presence of the reactive oxygen species mediator, mycolactone. To ensure regeneration of tissues in chronic ulcers, with proper and timely healing of the wounds, natural antioxidants that can combat the effects of induced reactive oxygen species in wound-healing cells ought to be investigated. Reactive oxygen species activity was determined in mycolactone-treated RAW 264.7 macrophages and the scavenging ability of the antioxidants (ascorbic acid, gallic acid, and green tea kombucha) against mycolactone-induced reactive oxygen species (superoxide anions) was assessed using fluorescein probe (DCF-DA) and nitroblue tetrazolium dye. Cytotoxicity of the antioxidants, mycolactone, and the protective effect of the antioxidants on the cells upon treatment with mycolactone were determined using the Alamar blue assay. The expression levels of endogenous antioxidant enzyme genes (superoxide dismutase, catalase, and glutathione peroxidase) in response to mycolactone-mediated reactive oxygen species were determined using RT-qPCR. Mycolactone induced the production of reactive oxygen species in RAW 264.7 macrophages, and the resulting superoxide anions were scavenged by some of the antioxidants. The selected endogenous antioxidant enzyme genes in the macrophages were upregulated in the presence of the antioxidants and mycolactone. The exogenously supplied ascorbic acid and green tea kombucha offered moderate protection to the macrophages against the toxicity of mycolactone. We conclude that the results provide insights into alternate and adjunct therapeutic approaches in Buruli ulcer treatment, which could significantly attenuate the toxicity of the pathogenic factor; mycolactone.

Dataset Information

Siegesbeckia pubescens Makino inhibits Pam₃CSK₄-induced inflammation in RAW 264.7 macrophages through suppressing TLR1/TLR2-mediated NF-?B activation.

Background

Methods

Results

Conclusions

Publications

<i>Siegesbeckia pubescens</i> Makino inhibits Pam<sub>3</sub>CSK<sub>4</sub>-induced inflammation in RAW 264.7 macrophages through suppressing TLR1/TLR2-mediated NF-κB activation.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets

Dataset Information

Siegesbeckia pubescens Makino inhibits Pam3CSK4-induced inflammation in RAW 264.7 macrophages through suppressing TLR1/TLR2-mediated NF-?B activation.

Background

Methods

Results

Conclusions

Publications

<i>Siegesbeckia pubescens</i> Makino inhibits Pam<sub>3</sub>CSK<sub>4</sub>-induced inflammation in RAW 264.7 macrophages through suppressing TLR1/TLR2-mediated NF-κB activation.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets

Siegesbeckia pubescens Makino inhibits Pam₃CSK₄-induced inflammation in RAW 264.7 macrophages through suppressing TLR1/TLR2-mediated NF-?B activation.