• Natural Product Sciences
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• v.22 no.1
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• pp.70-75
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• 2016

A phytoformula containing the root barks of Morus alba, the fructus of Schizandra sinensis and the roots of Asparagus cochinchinensis (MSA) was prepared as a potential new herbal remedy, and its therapeutic potential for alleviating inflammatory lung conditions was examined. For in vivo evaluation, an animal model of lipopolysaccharide (LPS)-induced acute lung injury (ALI) in mice was used. With oral administration of 6 - 60 mg/kg, MSA potently and dose-dependently inhibited bronchitis-like symptoms in acute lung injury induced by intranasal treatment of LPS as judged by the number of cells in the bronchoalveolar lavage fluid (BALF) and histological observation. The inhibitory potency was comparable with that of dexamethasone. For in vitro assay, the effects on the production of proinflammatory molecules in lung epithelial cells and alveolar macrophages were examined. Although MSA inhibited IL-6 production in IL-$1{\beta}$-treated lung epithelial cells (A549) only at a high concentration ($300{\mu}g/ml$), the formula strongly and concentration-dependently inhibited NO production in LPS-treated alveolar macrophages (MH-S) at $20-300{\mu}g/ml$. Based on all of these findings, the new phytoformula MSA is suggested to have the potential to control inflammatory lung diseases including bronchitis, at least in part, by inhibiting inducible nitric oxide synthase-catalyzed NO production.

• Journal of Physiology & Pathology in Korean Medicine
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• v.20 no.4
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• pp.966-972
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• 2006

The effect of water extract of Chungjogupae-tang (CJGPT) was investigated _on the growth of human lung carcinoma A549 cells. Methods: MTT assay and fluorescent microscope peformed to compare and examine the efficacy of CJGPT treatment on the cytostaticity of lung cancer cells in proportion to time and doses, and DAPI staining and Western blot analysis were used to examine their effect on apoptosis. In addition, the quantitative RT-PCR was used to examine to lung cancer cells growth, and Prostaglandin E2 activity were measured. Results: Exposure of A549 cells to CJGPT respited in the growth inhibition and apoptosis in a dose-dependent manner as measured by MTT assay and fluorescent microscope. The antiproliferative effect by CJGPT treatment in A549 cells was associated with morphological changes such as membrane shrinking and cell rounding up. CJGPT treatment resulted in an up-regulation of cyclin-dependent kinase inhibitor p21 (WAFl/CIPl) in a p53-independent fashion. We found that CJGPT treatment decreased the levels of cyclooxygenase (COX)-2 and inducible nitric oxide synthease (iNOS) expression without significant changes in the expression of COX-1 , which was correlated with a decrease in prostaglandin E2 (PGE2) synthesis. Conclusion: These findings suggested that CJGPT-induced inhibition of human lung carcinoma A549 cell growth was connected with the induction of apoptotic cell death and the results provided important new insights into the possible molecular mechanisms of the anti-cancer activity of CJGPT.

• Journal of Physiology & Pathology in Korean Medicine
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• v.20 no.4
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• pp.914-918
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• 2006

It was examined that the effect of fermented traditional wine made by using mycelium of Phellinus linteus on the expression of inflammation-related proteins in HepG2 cells. HepG2 cells were incubated with or without ertract of traditional wine (ETMP), then analyzed by microscopic observation, reverse transcription polymerase chain reaction (RT-PCR) and Western blot. The results of RT-PCR and Western blot analyses showed that the level of inducible nitric oxide synthase (iNOS), cyclooxygenase (COX)-2 and tumor necrosis factor $(TNF)-{\alpha}$ was induced by LPS, Dut the treatment of ETMP inhibited the expression of these proteins and its mRNAs. Besides, the results of Western blot analyses showed that the expression of nuclear $factor-{\kappa}Bp65$ and $inhibitory-{\kappa}B{\alpha}$ were also slightly affected by ETMP treatment. These results suggest that ETM P alleviate the expression of inflammation-related protein expressions and thus may be used as a functional alcoholic beverage.

• Journal of Physiology & Pathology in Korean Medicine
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• v.21 no.3
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• pp.642-646
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• 2007

It was examined that the effect of fermented traditional wine made by using mycelium of Phellinus linteus (TWPL) on the expression of inflammation-related proteins in rat liver. Levels of aspartate aminotransferase (AST) was significantly increased in the serum of ethanol-treated rats compared to normal. However, the level of AST showed no significant changes in the TWPL-treated rat compared normal. Slight histopathological changes of liver such as cloudy swelling, inflammatory cells infiltration, Kupffer cell reaction were demonstrated in the rats challenged with ethanol compared with normal. Fewer scores of these changes were observed in TWPL-treated rat with recovered glycogen in hepatocytes of whole hepatic lobule. The RT-PCR and Western analysis showed that the expression of inflammatory proteins such as cyclooxygenase-2, inducible nitric oxide synthase, tumor necrosis factor (TNF)-${\alpha}$ were decreased in the TWPL-treated rat compared with ethanol-treated ones. Immunohistochemical analysis showed that the expression of interleukin-lf and TNF-${\alpha}$ tended to decrease in TWPL-treated rat compared with ethanol-treated ones. These results suggest that TWPL may contains some protective agent for alcohol-induced liver injury through a regulating inflammation-related proteins.

• Food Science and Biotechnology
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• v.17 no.4
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• pp.745-750
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• 2008

Anti-inflammatory potential of the enzymatic extract prepared by Kojizyme (ECK), a component of brown seaweeds Ecklonia cava (Alariaceae, Phaeophyta) in vivo was investigated. For the application of mouse ear edema model, 12-O-tetradecanoylphorbol acetate (TPA) was used, a topical inducer of a long-lasting inflammatory response. Our results demonstrated that ECK inhibited ear edema when topically applied to mouse ear skin. In histological evaluation, the inhibition activity of ECK on TPA-induced inflammation is similar to that of dexamethasone, although less strong. In addition, the mRNA expression levels of IL-$1{\beta}$, IFN-$\gamma$, TNF-$\alpha$, and cyclooxygenase-2 (COX2) and the immunoreactivity to inducible nitric oxide synthase (iNOS) and COX2 expressed mainly in inflammatory cells were down-regulated by ECK. These results indicate that ECK has anti-inflammatory effects through the inhibition of Th1 cytokines and 2 inducers of inflammation in TPA-induced ear skin edema.

• The Korea Journal of Herbology
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• v.31 no.6
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• pp.37-44
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• 2016

Objective : This study aimed to evaluate the protective effect of Artemisiae Capillaris Herba (AC) in reflux esophagitis (RE) rats. Methods : The AC was measured antioxidant activity through in vitro experiments, such as total polyphenol and flavonoid contents, 1, 1-diphenyl-2-picrylhydrazyl (DPPH) and 2, 2'-azinobis-3-ethyl-benzothiazoline-6-sulfonic acid (ABTS) radical scavenging activity. Base on the results, we had conducted in vivo experiments. Rats were divided normal, control, AC treatment 50 mg/kg BW (AC50), and AC treatment 100 mg/kg BW (AC100) groups. AC were orally administered 2 h before the induction of RE. RE was induced by tie the pylorus and the transitional junction between the forestomach and the corpus in Sprague-Dawley rats. The rats were sacrificed 5 h after the surgery. We analyzed the expression of inflammatory related markers by western blot and observed the production of reactive oxygen species (ROS) and hematoxylin-eosin staining, Results : The $IC_{50}$ of AC for DPPH and ABTS were showed 12.60 and $33.32{\mu}g/m{\ell}$ respectively. In the RE rat, AC decreased inflammatory related markers, such as phosphorylated inhibitor of ${\kappa}B{\alpha}$, nuclear factor-kappa B, cyclooxygenase-2, inducible nitric oxide synthase, and tumor necrosis factor alpha. Also, AC reduced the increased reactive oxygen species in serum. The anti-inflammatory effect of AC appeared to be partially mediated through the inhibition of ROS. Also, AC markedly ameliorated esophageal mucosa damage via the inhibition of protein expression related to inflammation. Conclusions : Therefore, these results suggest that AC would be used as a therapeutic material in protection and/or treatment for reflux esophagitis.

• Archives of Pharmacal Research
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• v.24 no.6
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• pp.607-612
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• 2001

Endogenous carbon monoxide (CO) shares with nitric oxide (NO) a role as a putative neural messenger in the brain. Both gases are believed to modulate CNS function via an increase in cytoplasmic cGMP concentrations secondary to the activation of soluble guanylate cyclase (sGC). Recently CO and NO were proposed as a possible mediator of febrile response in hypothalamus. NO has been reported to activate both the constitutive and inducible isoform of the cyclooxygenase (COX). Thus, we investigated whether CO arising from heme catabolism by heme oxygenate (HO) is involved in the febrile response via the activation of COX in the hypothalamus. $PGE_2$ which is a final mediator of febrile response released from primary cultured hypothalamic cells was taken as a marker of COX activity. $PGE_2$ concentration was measured with EIA kits. Exogenous CO (CO-saturated medium) and hemin (a substrate and potent inducer of HO) evoked an increase in $PGE_2$ release from hypothalamic cells, and these effects were blocked by methylene blue (an inhibitor of sGC). And membrane permeable cGMP analogue, dibutyryl-cGMP elicited significant increases in $PGE_2$release. These results suggest that there may be a functional link between HO and COX enzymatic activities. The gaseous product of hemin through the HO pathway, CO, might play a role through the modulation of the COX activity in the hypothalamus.

• The Korean Journal of Physiology and Pharmacology
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• v.23 no.6
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• pp.493-499
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• 2019

Macrophage-associated inflammation is crucial for the pathogenesis of diverse diseases including metabolic disorders. Rhodanthpyrone (Rho) is an active component of Gentiana rhodantha, which has been used in traditional Chinese medicine to treat inflammation. Although synthesis procedures of RhoA and RhoB were reported, the biological effects of the specific compounds have never been explored. In this study, the anti-inflammatory activity and mechanisms of action of RhoA and RhoB were studied in lipopolysaccharide (LPS)-stimulated macrophages. Pretreatment with RhoA and RhoB decreased inducible nitric oxide synthase and cyclooxygenase-2 expressions in RAW 264.7 cells and in thioglycollate-elicited mouse peritoneal macrophages. In addition, it downregulated transcript levels of several inflammatory genes in LPS-stimulated RAW 264.7 cells, including inflammatory cytokines/chemokines (Tnfa, Il6, and Ccl2) and inflammatory mediators (Nos2 and Ptgs2). Macrophage chemotaxis was also inhibited by treatment with the compounds. Mechanistic studies revealed that RhoA and RhoB suppressed the nuclear factor $(NF)-{\kappa}B$ pathway, but not the canonical mitogen activated protein kinase pathway, in LPS-stimulated condition. Moreover, the inhibitory effect of RhoA and RhoB on inflammatory gene expressions was attenuated by treatment with an $NF-{\kappa}B$ inhibitor. Our findings suggest that RhoA and RhoB play an anti-inflammatory role at least in part by suppressing the $NF-{\kappa}B$ pathway during macrophage-mediated inflammation.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.31 no.1
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• pp.12-21
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• 2018

Objectives : Inflammation is one of the self-protective abilities against tissue injury, and it has clinical symptoms like redness, heat, swelling, pain, and loss of function. The purpose of this study is to examine inhibitory effects of Naetakchunkeum-san (NTCKS) on nitric oxide (NO), Prostaglandin E2 (PGE2), inducible NOS (iNOS), cyclooxygenase-2 (COX-2), and phosphorylation of extracellular signal-regulated kinase 1/2 (ERK1/2), which play a major role in inflammatory response. Methods : The experiment was performed using Raw 264.7 cells pretreated with NTCKS extracts. Cell viability was determined by MTT assay. To evaluate anti-inflammatory effects of NTCKS, we examined NO and $PGE_2$ production in LPS-induced macrophages. We also investigated effects of NTCKS on iNOS, Cox-2, and ERK1/2 expression using western blot. Results : In MTT assay, no cytotoxicity of NTCKS (50, 100, 150, $200{\mu}g/ml$) on RAW 264.7 cell was found. LPS-induced NO production was decreased after treatment with NTCKS (150, $200{\mu}g/ml$)(p<0.05). $PGE_2$ was decreased after treatment with NTCKS (150, $200{\mu}g/ml$)(p<0.05). NTCKS inhibited LPS-induced expressions of iNOS and COX-2 in a dose-dependent manner. Increased phosphorylation of ERK1/2 by LPS was decreased by NTCKS in a dose-dependent manner. Conclusions : According to above experiments, NTCKS may be applied to inflammatory diseases such as atopic dermatitis, rheumatoid arthritis, and inflammatory bowel disease.

• Archives of Pharmacal Research
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• v.27 no.7
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• pp.757-762
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• 2004

The protective adaptive response to electrophiles and reactive oxygen species is mediated by the induction of phase II detoxifying genes including glutathione S-transferases (GSTs). NF-E2-related factor-2 (Nrf2) phosphorylation by protein kinase C (PKC) is a critical event for its nuclear translocation in response to oxidative stress. Previously, we have shown that peroxynitrite plays a role in activation of Nrf2 and Nrf2 binding to the antioxidant response element (ARE) via the pathway of phosphatidylinositol 3-kinase (PI3-kinase) and that nitric oxide synthase in hepatocytes is required for GSTA2 induction. In view of the importance of PKC and Pl3-kinase in Nrf2-mediated GST induction, we investigated the role of these kinases in peroxynitrite formation for GSTA2 induction by oxidative stress and determined the relationship between PKC and PI3-kinase. Although PKC activation by phorbol 12-myristate-13-acetate (PMA) did not increase the extents of constitutive and inducible GSTA2 expression, either PKC depletion by PMA or PKC inhibition by staurosporine significantly inhibited GSTA2 induction by tert-butylhydroquinone (t-SHa) a prooxidant chemical. Therefore, the basal PKC activity is req- uisite for GSTA2 induction. 3-Morpholinosydnonimine (SIN-1), which decomposes and yields peroxynitrite, induced GSTA2, which was not inhibited by PKC depletion, but slightly enhanced by PKC activation, suggesting that PKC promotes peroxynitrite formation for Nrf2-mediated GSTA2 induction. Treatment of cells with S-nitroso-N-acetyl-penicillamine (SNAP), an exogenous NO donor, in combination with t-BHQ may produce peroxynitrite. GSTA2 induction by SNAP ＋ t-BHQ was not decreased by PKC depletion, but rather enhanced by PKC activation, showing that the activity of PKC might be required for peroxynitrite formation. LY294002 a P13-kinase inhibitor blocked GSTA2 induction by t-BHQ, which was reversed by PMA-induced PKC activation. These results provide evidence that PKC may playa role in formation of peroxynitrite that activates Nrf2 for GSTA2 induction and that PKC may serve an activator for GSTA2 induction downstream of PI3-kinase.