Cargando…

Anti-inflammatory and antioxidant effects of MOK, a polyherbal extract, on lipopolysaccharide-stimulated RAW 264.7 macrophages

MOK, a pharmacopuncture medicine consisting of 10 herbs, has a long history as treatment for various inflammatory conditions. To investigate the mechanisms of action of MOK, its anti-inflammatory and antioxidative effects were assessed in RAW 264.7 macrophages stimulated by lipopoly-saccharide (LPS)...

Descripción completa

Detalles Bibliográficos
Autores principales: Hwang, Ji Hye, Ma, Jun Nan, Park, Jong Hun, Jung, Hyo Won, Park, Yong-Ki
Formato: Online Artículo Texto
Lenguaje:English
Publicado: D.A. Spandidos 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6257867/
https://www.ncbi.nlm.nih.gov/pubmed/30365058
http://dx.doi.org/10.3892/ijmm.2018.3937
_version_ 1783374410323853312
author Hwang, Ji Hye
Ma, Jun Nan
Park, Jong Hun
Jung, Hyo Won
Park, Yong-Ki
author_facet Hwang, Ji Hye
Ma, Jun Nan
Park, Jong Hun
Jung, Hyo Won
Park, Yong-Ki
author_sort Hwang, Ji Hye
collection PubMed
description MOK, a pharmacopuncture medicine consisting of 10 herbs, has a long history as treatment for various inflammatory conditions. To investigate the mechanisms of action of MOK, its anti-inflammatory and antioxidative effects were assessed in RAW 264.7 macrophages stimulated by lipopoly-saccharide (LPS). RAW 264.7 cells were treated with different concentrations of MOK extract for 30 min prior to stimulation with or without LPS for the indicated times. Nitric oxide (NO) production was measured using Griess reagent, while the mRNA levels of inflammatory cytokines, tumor necrosis factor (TNF)-α, interleukin (IL)-1β, IL-6 and the antioxidant enzymes Mn superoxide dismutase and heme oxygenase-1, were determined using reverse transcription-polymerase chain reaction analysis. Western blotting was used to determine the protein expression of inducible nitric oxide synthase (iNOS), cyclooxygenase (COX)-2, superoxide dismutase (SOD)2, catalase (CAT) and heme oxygenase-1 (HO-1), and the phosphorylation of mitogen-activated protein kinases (MAPKs), including ERK1/2, JNK and p38. Western blotting and immunocytochemistry were used to observe the nuclear expression of nuclear factor (NF)-κB p65. Additionally, reactive oxygen species (ROS) and prostaglandin (PG)E(2) production were determined using the ROS assay and an enzyme immunoassay. With MOK treatment, there was a notable decrease in NO and PGE(2) production induced by LPS in RAW 264.7 cells by downregulation of iNOS and COX-2 mRNA and protein expression. Furthermore, with MOK treatment, there was a decrease in the mRNA expression levels of TNF-α, IL-1β and IL-6, as well as in the phosphorylation of ERK, JNK and p38 MAPK, by blocking the nuclear translocation of NF-κB p65 in LPS-stimulated cells. In addition, MOK treatment led to an increase in the antioxidant enzymes SOD, CAT and HO-1 in LPS-stimulated cells, with a concomitant decrease in ROS generation. These results indicate that the inflammatory responses in activated macrophages are inhibited by MOK through downregulation of the transcription levels of inflammatory mediators and inhibition of the MAPK/NF-κB pathway. Moreover, MOK protects against oxidative damage by upregulating the expression of antioxidant enzymes and generating ROS scavengers.
format Online
Article
Text
id pubmed-6257867
institution National Center for Biotechnology Information
language English
publishDate 2019
publisher D.A. Spandidos
record_format MEDLINE/PubMed
spelling pubmed-62578672018-11-29 Anti-inflammatory and antioxidant effects of MOK, a polyherbal extract, on lipopolysaccharide-stimulated RAW 264.7 macrophages Hwang, Ji Hye Ma, Jun Nan Park, Jong Hun Jung, Hyo Won Park, Yong-Ki Int J Mol Med Articles MOK, a pharmacopuncture medicine consisting of 10 herbs, has a long history as treatment for various inflammatory conditions. To investigate the mechanisms of action of MOK, its anti-inflammatory and antioxidative effects were assessed in RAW 264.7 macrophages stimulated by lipopoly-saccharide (LPS). RAW 264.7 cells were treated with different concentrations of MOK extract for 30 min prior to stimulation with or without LPS for the indicated times. Nitric oxide (NO) production was measured using Griess reagent, while the mRNA levels of inflammatory cytokines, tumor necrosis factor (TNF)-α, interleukin (IL)-1β, IL-6 and the antioxidant enzymes Mn superoxide dismutase and heme oxygenase-1, were determined using reverse transcription-polymerase chain reaction analysis. Western blotting was used to determine the protein expression of inducible nitric oxide synthase (iNOS), cyclooxygenase (COX)-2, superoxide dismutase (SOD)2, catalase (CAT) and heme oxygenase-1 (HO-1), and the phosphorylation of mitogen-activated protein kinases (MAPKs), including ERK1/2, JNK and p38. Western blotting and immunocytochemistry were used to observe the nuclear expression of nuclear factor (NF)-κB p65. Additionally, reactive oxygen species (ROS) and prostaglandin (PG)E(2) production were determined using the ROS assay and an enzyme immunoassay. With MOK treatment, there was a notable decrease in NO and PGE(2) production induced by LPS in RAW 264.7 cells by downregulation of iNOS and COX-2 mRNA and protein expression. Furthermore, with MOK treatment, there was a decrease in the mRNA expression levels of TNF-α, IL-1β and IL-6, as well as in the phosphorylation of ERK, JNK and p38 MAPK, by blocking the nuclear translocation of NF-κB p65 in LPS-stimulated cells. In addition, MOK treatment led to an increase in the antioxidant enzymes SOD, CAT and HO-1 in LPS-stimulated cells, with a concomitant decrease in ROS generation. These results indicate that the inflammatory responses in activated macrophages are inhibited by MOK through downregulation of the transcription levels of inflammatory mediators and inhibition of the MAPK/NF-κB pathway. Moreover, MOK protects against oxidative damage by upregulating the expression of antioxidant enzymes and generating ROS scavengers. D.A. Spandidos 2019-01 2018-10-16 /pmc/articles/PMC6257867/ /pubmed/30365058 http://dx.doi.org/10.3892/ijmm.2018.3937 Text en Copyright: © Hwang et al. This is an open access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License (https://creativecommons.org/licenses/by-nc-nd/4.0/) , which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made.
spellingShingle Articles
Hwang, Ji Hye
Ma, Jun Nan
Park, Jong Hun
Jung, Hyo Won
Park, Yong-Ki
Anti-inflammatory and antioxidant effects of MOK, a polyherbal extract, on lipopolysaccharide-stimulated RAW 264.7 macrophages
title Anti-inflammatory and antioxidant effects of MOK, a polyherbal extract, on lipopolysaccharide-stimulated RAW 264.7 macrophages
title_full Anti-inflammatory and antioxidant effects of MOK, a polyherbal extract, on lipopolysaccharide-stimulated RAW 264.7 macrophages
title_fullStr Anti-inflammatory and antioxidant effects of MOK, a polyherbal extract, on lipopolysaccharide-stimulated RAW 264.7 macrophages
title_full_unstemmed Anti-inflammatory and antioxidant effects of MOK, a polyherbal extract, on lipopolysaccharide-stimulated RAW 264.7 macrophages
title_short Anti-inflammatory and antioxidant effects of MOK, a polyherbal extract, on lipopolysaccharide-stimulated RAW 264.7 macrophages
title_sort anti-inflammatory and antioxidant effects of mok, a polyherbal extract, on lipopolysaccharide-stimulated raw 264.7 macrophages
topic Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6257867/
https://www.ncbi.nlm.nih.gov/pubmed/30365058
http://dx.doi.org/10.3892/ijmm.2018.3937
work_keys_str_mv AT hwangjihye antiinflammatoryandantioxidanteffectsofmokapolyherbalextractonlipopolysaccharidestimulatedraw2647macrophages
AT majunnan antiinflammatoryandantioxidanteffectsofmokapolyherbalextractonlipopolysaccharidestimulatedraw2647macrophages
AT parkjonghun antiinflammatoryandantioxidanteffectsofmokapolyherbalextractonlipopolysaccharidestimulatedraw2647macrophages
AT junghyowon antiinflammatoryandantioxidanteffectsofmokapolyherbalextractonlipopolysaccharidestimulatedraw2647macrophages
AT parkyongki antiinflammatoryandantioxidanteffectsofmokapolyherbalextractonlipopolysaccharidestimulatedraw2647macrophages