• The Journal of Korean Medicine
• /
• v.41 no.4
• /
• pp.12-26
• /
• 2020

Objectives: The aim of this study is to investigate the mechanisms involved in the anti-inflammatory and anti-tumor effects of Rhus verniciflua Stokes (RVS) on PMA-differentiated human monocytic leukemia THP-1 cells. Methods: Cells were treated with various concentrations of RVS decoction (0-300㎍/ml) for 24, 48, and 72h. Cell viability was evaluated by MTS/PMS assay. The expressions of MMP-2, MMP-9, TIMP-1, TIMP-2, iNOS and COX-2 mRNA and proteins were measured using RT-PCR and western blotting, respectively. Results: RVS suppressed expression of MMP-2 and MMP-9 mRNA. It also down-regulated iNOS and COX-2 mRNA and protein expression. RVS inhibited NF-𝜅B p65 activity and the phosphorylation of Akt and MAPK (ERK and p38 MAPK). Instead, the phosphorylation of JNK is increased at a very low concentration but decreased at higher concentrations. Conclusion: RVS is regarded to inhibit the expression of MMP and TIMP as well as iNOS and COX-2 gene expression via directly inhibiting the activation of NF-𝜅B and phosphorylation of MAPK pathway in THP-1 cells. This suggests RVS have potential to be used as a therapeutic agent for acute myeloid leukemia (AML).

• Journal of Physiology & Pathology in Korean Medicine
• /
• v.26 no.3
• /
• pp.293-300
• /
• 2012

Inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) are important inflammatory mediators implicated in pathogenesis of inflammation and certain types of human cancers. The present study was designed to determine whether Red Ginseng (RG) could modulate $I{\kappa}B$-kinase, iNOS and COX-2 gene expression and immune responses in RAW 264.7 macrophages stimulated with lipopolysaccharide (LPS). RG extract suppressed the expression of LPS-induced $I{\kappa}B$, iNOS, COX-2, and immune responses in a dose-dependent manner. It also showed an anti-inflammatory effect by inhibiting NF-${\kappa}B$ immune response induced by LPS treatment. Inhibitory effect of RG on LPS-induced inflammation was mediated by suppressed phosphorylation of ERK, JNK and p38 through the regulation of the mitogen-activated protein kinase (MAPK) pathway leading to a decreased production of NO, iNOS, COX-2 and NF-${\kappa}B$. The results implied the role of RG as an inflammation regulator and its possible application for curing inflammatory diseases.

• Archives of Pharmacal Research
• /
• v.26 no.10
• /
• pp.832-839
• /
• 2003

Activated macrophages express inducible isoforms of nitric oxide synthase (iNOS) and cyclooxygenase (COX-2), and produce excessive amounts of nitric oxide (NO) and prostaglandin E$_2$ (PGE$_2$), which play key roles in the processes of inflammation and carcinogenesis. The root of Paeonia lactiflora Pall., and the root cortex of Paeonia suffruticosa Andr., are important Chinese crude drugs used in many traditional prescriptions. 1,2,3,4,6-penta-O-galloyl-$\beta$-D-glucose (PGG) is a major bioactive constituent of both crude drugs. PGG has been shown to possess potent anti-oxidant, anti-mutagenic, anti-proliferative and anti-invasive effects. In this study, we examined the inhibitory effects of 1,2,3,4,6-penta-O-galloyl-$\beta$-D-glucose (PGG) isolated from the root of Paeonia lactiflora Pall. on the COX-2 and iNOS activity in LPS-activated Raw 264.7 cells, COX-1 in HEL cells. To investigate the structure-activity relationships of gallate and gallic acid for the inhibition of iNOS and COX-2 activity, we also examined (-)-epigallocatechin gallate (EGCG), gallic acid, and gallacetophenone. The results of the present study indicated that PGG, EGCG, and gallacetophenone treatment except gallic acid significantly inhibited LPS-induced NO production in LPS-activated macrophages. All of the four compounds significantly inhibited COX-2 activity in LPS-activated macrophages. Among the four compounds examined, PGG revealed the most potent in both iNOS ($IC_{50}$ = 18 $\mu\textrm{g}/mL$) and COX-2 inhibitory activity (PGE$_2$: $IC_{50}$ = 8 $\mu\textrm{g}/mL$ and PGD$_2$: $IC_{50}$ = 12 $\mu\textrm{g}/mL$), respectively. Although further studies are needed to elucidate the molecular mechanisms and structure-activity relationship by which PGG exerts its inhibitory actions, our results suggest that PGG might be a candidate for developing anti-inflammatory and cancer chemopreventive agents.

• The Korea Journal of Herbology
• /
• v.30 no.6
• /
• pp.77-82
• /
• 2015

Objectives : This study was designed to investigate of the anti-inflammatory effects of Schisandra chinensis seed oil(SSO) on the production of pro-inflammatory substances in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages.Methods : SSO was measured the production of pro-inflammatory factor (NO, PGE2, IL-1β iNOS and, COX-2) in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. we used the following methods : cell viability assay, Griess reagent assay, enzyme-linked immunosorbent assay, Western blotting analysis.Results : The cell viability of SSO(0∼500 μl/mL) processing group was 96.9% and the processing of SSO didn't have an effect on the cytotoxicity. The inhibitory effect of the nitric oxide (no) production of SSO(500 μg/mL, 50 μg/mL, 10 μg/mL) was each 70.3%, 37.6% and 26.5%. IL-1β production inhibition ability of SSO(500 μg/mL, 100 μg/mL) was each 49.88% and 48.8%. PGE2 production inhibition ability of SSO(500 μg/mL, 100 μg/mL) was each 49.88% and 73.1%, 70.5%. By using SSO, it experimented about iNOS protein expression inhibition ability, that is the NO production enzyme. iNOS protein expression increased in the group processing LPS independently. iNOS protein expression decreased in the group processing SSO together. The expression of the COX-2 protein decreased 89.6%, 81.8% in the group processing SSO. The significance was in the relationship with NO formation inhibition with the relationship with the PGE₂ formation inhibition and iNOS protein, it confirmed in SSO with the COX-2 protein.Conclusions : Stimulation of the RAW 264.7 cells with LPS caused an elevated production of nitric oxide (NO), IL-1β and PGE₂ which was markedly inhibited by the pretreatment with SSO without causing any cytotoxic effects. The reduced expressions of iNOS protein were consistent with the reductions in NO production in the culture media. SSO may be useful for the treatment of various inflammatory diseases.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
• /
• v.26 no.4
• /
• pp.70-80
• /
• 2013

Objectives : Haliotidis Concha has been used to treat various human diseases such as liver dysfunction and inflammatory disorder. Although it has been shown the effects of Haliotidis Concha on the various diseases, it has almost not been studied about the anti-inflammatory effects of the Haliotidis Concha and its mechanisms. Methods : This research investigated the effects of the Haliotidis Concha ethanol extract (HCE) on the production of nitric oxide (NO) and prostaglandin $E_2$ ($PGE_2$) as well as tumor necrosis factor-alpha (TNF-${\alpha}$). The protein expressions of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) were assayed by immunoblot analyses, and the productions of NO, $PGE_2$ and TNF-${\alpha}$ were assessed by ELISA. Results : Haliotidis Concha decreased the production of NO and $PGE_2$, and inhibited the expression iNOS and COX-2 proteins in a concentration-dependent manner in LPS-treated Raw 264.7 cells. HCE suppressed the ability of LPS to activate the signaling pathways of nuclear factor kappa B (NF-${\kappa}B$) as indicated by HCE inhibited nuclear NF-${\kappa}B$ level and I-${\kappa}B{\alpha}$ phosphorylation. Also, HCE inhibited mitogen-activated protein kinases (MAPKs). Conclusions : HCE repressed the production of LPS-inducible NO, $PGE_2$ and TNF-${\alpha}$, which may be mediated by inhibition of NF-${\kappa}B$ translocation. This study suggest the use for the treatment of acute inflammatory disorders.

• Journal of Life Science
• /
• v.26 no.12
• /
• pp.1422-1430
• /
• 2016

This study was performed to investigate the modulatory effects of two prototypes of Panax ginseng saponin fractions, 20(S)-protopanaxadiol saponins (PDS) and 20(S)-protopanaxatriol saponins (PTS), on the induction of inflammatory mediators in lipopolysaccharide (LPS)-treated RAW264.7 murine macrophage cells. For this purpose, RAW264.7 cells were treated with LPS ($10{\mu}g/ml$) before, after, or simultaneously with PDS or PTS ($150{\mu}g/ml$), and the released level of nitric oxide (NO) and expression levels of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) were evaluated. When RAW264.7 cells were treated with LPS and ginseng saponin fractions simultaneously for 24 hr, PTS, compared to PDS, more strongly attenuated the NO production induced by LPS treatment. When the cells were pretreated with LPS for 2 hr followed by PDS or PTS treatment for 24 hr, both ginseng saponins strongly reduced NO release. The pretreatment of RAW264.7 cells with PDS or PTS for 2 hr followed by LPS treatment for 24 hr significantly attenuated the LPS-induced production of NO. PTS showed stronger inhibitory potency to NO generation than PDS. Our western blot experiment showed that both PDS and PTS ($150{\mu}g/ml$) also significantly down-regulated the expressions of iNOS and COX-2 induced by LPS treatment. Our results suggest that both PDS and PTS possess strong protective effects against LPS-stimulated inflammation and that their protective effects are mediated by the suppression of NO synthesis via down-regulation of pro-inflammatory enzymes, iNOS, and COX-2 in the RAW264.7 cells.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
• /
• v.23 no.2
• /
• pp.13-26
• /
• 2010

Objectives : The present study was conducted to evaluate the anti-inflammatory effects of the Gamroeum water extracts (GRE) in vivo and in vitro. Methods : The effects of GRE on anti-inflammation were measured by production of NO, $PGE_2$ (Prostaglandin $E_2$), iNOS (inducible Nitric Oxide Synthase), COX-2, $NF{\kappa}B$ (Nuclear Factor kappa B), TNF-$\alpha$ (Tumor Necrosis Factor-alpha) and IL-$1{\beta}$ (Interleukin-$1{\beta}$), IL-6 in Raw 264.7 macrophage cells stimulated with LPS. Results : 1. In machrophage cells, LPS displayed significant stimulatory effects on the production of NO and $PGE_2$. However, GRE showed significant inhibitory effects on NO and $PGE_2$ release. The level of NO and $PGE_2$ was decreased by GRE in a concentration dependent manner as compared with LPS only group. 2. Immunoblot analysis verified that LPS stimulation significantly increased the iNOS and COX-2 protein level, but GRE suppressed the induction of iNOS and COX-2 protein at a concentration dependent manner. 3. GRE reduced the elevated production of TNF-$\alpha$, IL-$1{\beta}$ and IL-6 by LPS. Moreover, the inhibitory effects of GRE was occurred in a dose-dependent manner. 4. GRE significantly reduced the expression of NF-${\kappa}B$ protein in nuclear fraction. 5. GRE effectively inhibited the increases of hind paw skin thicknesses and inflammatory cell infiltrations induced by carrageenan treatment. It, therefore, considered that GRE will be favorably inhibited the acute edematous inflammations. Conclusions : These results indicated that GRE could have anti-inflammatory capacity by inhibiting the production of NO, $PGE_2$ and cytokines in vitro and by reducing the formation of carrageenan-induced paw edema in vivo. Moreover, inhibitory effects of GRE on the macrophage activation were attributable to the reduction of some of inflammatory factors by inhibiting iNOS and COX-2 through the suppression of NF-${\kappa}B$.

• Journal of Physiology & Pathology in Korean Medicine
• /
• v.26 no.4
• /
• pp.483-490
• /
• 2012

This study was designed to identify the effects of Polygoni cuspidati Radix(PCR) on the generation of superoxide anion radicals (${\cdot}O_2{^-}$), nitric oxide (NO), peroxynitrite ($ONOO^-$) in the renal epithelial cells of mouse(LLC-$PK_1$). The effects of PCR on the expression of inflammation-related proteins, IKK-${\alpha}$, phospho-$I{\kappa}B-{\alpha}$, NF-${\kappa}B$ (p50, p65), COX-2, iNOS, IL-$1{\beta}$, VCAM-1, were examined by western blotting. For this study, the fluorescent probes, namely dihydrorhodamine 123 (DHR 123), 2',7'-dichloro dihydrofluorescein diacetate (DCFDA), 4,5-diaminofluorescein (DAF-2) were used. Protein expression levels of IKK-${\alpha}$, phospho-$I{\kappa}B-{\alpha}$, NF-${\kappa}B$ (p50, p65), COX-2, iNOS, IL-$1{\beta}$, VCAM-1 were assayed by western blot. PCR reduced $H_2O_2$-induced cell death dose-dependently. It inhibited the generation of ${\cdot}O_2{^-}$, NO, $ONOO^-$ and $PGE^2$ in the $H_2O_2$-treated LLC-PK1 cells in vitro. PCR inhibited the espression of IKK-${\alpha}$, phospho-$I{\kappa}B-{\alpha}$, COX-2, iNOS, IL-$1{\beta}$ and VCAM-1 genes by means of decreasing the NF-${\kappa}B$ activation. These results suggest that PCR is an effective NO, ${\cdot}O_2{^-}$, $ONOO^-$ scavenger, and this substance recommended to be applied in treatment for the inflammatory process and inflammation-related disease.

• Korean Journal of Medicinal Crop Science
• /
• v.26 no.1
• /
• pp.8-18
• /
• 2018

Background: This present study was conducted to evaluate the anti-inflammatory and immune regulatory effects of Aucklandia lappa Decne (AL). Methods and Results: We measured cytotoxicity, nitric oxide (NO) content, mRNA expression (iNOS, IL-1${\alpha}$, IL-$1{\beta}$, and TNF-${\alpha}$), protein expression (iNOS, COX-2, and $I{\kappa}B$) and phagocytic activity in RAW264.7 cells. Male BALB/c mice were fed 100 mg/kg AL (Aucklandia lappa Decneon 70% ethanol extract) and 250 mg/kg AL for 4 weeks; thereafter, we observed B/T or $CD4^+/CD8^+$ lymphocyte subpopulation change, and expression patterns of $CD4^+$ and $CD8^+$ lymphocytes by immunohistochemical staining in mouse splenocytes and/or thymocytes. To determine the experimental concentration of AL, cell viability was measured by MTT assay and tested at $12.5{\mu}g/m{\ell}$ or less. AL inhibited the levels of NO, lymphokine production (IL-$1{\beta}$, and TNF-${\alpha}$), and mRNA (iNOS, IL-1${\alpha}$, IL-$1{\beta}$, and TNF-${\alpha}$) and protein (iNOS, and COX-2) expression. Additionally, the levels of $I{\kappa}B$, phagocytic activity, and splenic and thymic T lymphocytes, especially $T_H$ and $T_C$ cells were significantly increased in AL administered mice. The immuno-reactive density of $CD4^+$ and $CD8^+$ lymphocytes was stronger in AL groups than in the normal group. AL stimulated NO, iNOS, and COX-2, and regulated IL-1${\alpha}$, IL-$1{\beta}$, TNF-${\alpha}$, and $I{\kappa}B$ in macrophages treated with LPS (lipopolysaccharide). In addition, AL increased the phagocytic activity of macrophages and the immunity of mouse T ($T_H$, and $T_C$) cells. Conclusions: These results suggested that AL might show anti-inflammatory activity via the suppression of various inflammatory markers and immuno-regulatory activity.

• Biomedical Science Letters
• /
• v.26 no.4
• /
• pp.267-274
• /
• 2020

The larva of Protaetia brevitarsis Lewis (P. brevitarsis), edible insect, is traditionally consumed as alternative source of nutrients and has various health benefits. However, the exact pharmaceutical effects of P. brevitarsis on inflammatory response are still not well understood. Thus, we investigated the anti-inflammatory effects and mechanisms of P. brevitarsis in lipopolysaccharide (LPS)-stimulated RAW264.7 cells. We investigated the effects of P. brevitarsis on the expression levels of inflammatory-related genes, including inflammatory cytokines, prostaglandin E2 (PGE2), cyclooxygenase (COX)-2 and inducible nitric oxide synthase (iNOS) in LPS-stimulated RAW264.7 cells. To understand the anti-inflammatory mechanism of P. brevitarsis, we explored the regulatory effect of P. brevitarsis on nuclear factor (NF)-κB and caspase-1 activation. The findings of this study demonstrated that P. brevitarsis inhibits the LPS-induced inflammatory cytokine and PGE2 levels, as well as COX-2 and iNOS expression. Moreover, we confirmed that the anti-inflammatory effect of P. brevitarsis occurs via suppression of the activation of NF-κB and caspase-1. Conclusively, these findings provide experimental evidence that P. brevitarsis may be useful candidate for the treatment of inflammatory-related diseases.