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