• Journal of Acupuncture Research
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• v.20 no.5
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• pp.93-106
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• 2003

Objective : To investigate the possible molecular mechanism(s) of melittin as a candidate of anti-cancer drug, we examined the effects of the compound on the growth of human lung carcinoma cell line A549. Methods: MTT, morphological changes, DAPI staining, Western blot, RT-PCR and in vitro prostaglandin E2 (PGE2) accumulation assays were performed. Results: The anti-proliferative effect by melittin treatment in A549 cells was associated with morphological changes such as membrane shrinking and cell rounding up. Melittin induced apoptotic cell death in a concentration-dependent manner, which was associated with inhibition or degradation of apoptotic target proteins such as ${\beta}$-catenin, poly(ADP-ribose) polymerase(PARP) and phospholipase $C-{\gamma}1(PLC-{\gamma}1)$. Melittin treatment inhibited the expression of cyclooxygenase-2(COX-2) and accumulation of PGE2 in aconcentration-dependent fashion. In addition, Melittin treatment induced the down-regulation of telomerase reverse transcriptase(hTERT) and proto-oncogene c-myc expression of A549 cells. Conclusions: Taken together, these findings suggest that melittin-induced inhibition of human lung cancer cell proliferation is associated with the induction of apoptotic cell death via regulation of several major growth regulatory gene products, and melittin may have therapeutic potential in human lung cancer.

• Preventive Nutrition and Food Science
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• v.17 no.1
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• pp.22-28
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• 2012

Melaleuca leucadendron L. has been used as a tranquilizing, sedating, evil-dispelling and pain-relieving agent. We examined the effects of M. leucadendron L. extracts on oxidative stress and inflammation. M. leucadendron L. was extracted with methanol (MeOH) and then fractionated with chloroform ($CHCl_3$) and butanol (BuOH). Antioxidant activity of the MeOH extract and BuOH fraction were higher than that of both ${\alpha}$-tocopherol and butyrated hydroxytoluene (BHT). Total phenol content in the extracts of M. leucadendron L., especially the BuOH fraction, well correlated with the antioxidant activity. The anti-inflammatory activity of BuOH extracts were investigated by lipopolysaccharide (LPS)-induced nitric oxide (NO) and prostaglandin $E_2$ ($PGE_2$) production, and cyclooxygenase-2 (COX-2) expression in RAW 264.7 macrophages. The BuOH fraction significantly inhibited LPS-induced NO and $PGE_2$ production. Furthermore, BuOH extract of M. leucadendron L. inhibited the expression of COX-2 and iNOS protein without an appreciable cytotoxic effect on RAW264.7 cells. The extract of M. leucadendron L. also suppressed the phosphorylation of inhibitor ${\kappa}B{\alpha}$ ($I{\kappa}B{\alpha}$) and its degradation associated with nuclear factor-${\kappa}B$ (NF-${\kappa}B$) activation. Furthermore, BuOH fraction inhibited LPS-induced NF-${\kappa}B$ transcriptional activity in a dose-dependent manner. These results suggested that M. leucadendron L. could be useful as a natural antioxidant and anti-inflammatory resource.

• Biomolecules & Therapeutics
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• v.16 no.3
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• pp.277-285
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• 2008

Panax ginseng C.A. Mayer (Araliaceae, P. ginseng) has been used for the enhancement of vascular and immune functions in Korea and Japan for a long time. Ginsenoside $Rb_1$ and $Rg_3$ isolated from P. ginseng head-part butanolic extract (PGHB) were investigated for anti-inflammatory activity. Ginsenosides and PGHB did not affect the cell viability within $0\;-\;100\;{\mu}g/ml$ concentration to RAW 264.7 murine macrophage cells. Ginsenosides and PGHB inhibited partly lipopolysaccharide (LPS)-induced nitrite production in a dose-dependent manner. The ginsenosides and PGHB showed partially chemical nitric oxide (NO) quenching (maximum 40%) in the cell-free system. Also, ginsenoside $Rb_1$ and $Rg_3$ inhibited markedly approximately 74 and 54% of inducible nitric oxide synthase (iNOS) mRNA transcription from LPS-induced RAW 264.7 cells. Taken together, the inhibitory effect of ginsenosides and PGHB on NO production did not occur as a result of cell viability, but was caused by both the chemical NO quenching and the regulation of iNOS. Additionally, the ginsenoside $Rb_1$ and PGHB inhibited prostaglandin $E_2$ ($PGE_2$) synthesis in a concentration-dependent manner, showed approximately 70-98% inhibition at $100\;{\mu}g/ml$ concentration. And the treatment with ginsenosides and PGHB attenuated partially LPS-upregulated cyclooxygenase-2 (COX-2) gene transcription. Ginsenoside $Rg_3$ suppressed LPS-stimulated interleukin-6 (IL-6) level to the basal in RAW 264.7 cells. From these results, ginsenoside $Rb_1,\;Rg_3$, and PGHB may be useful for the relief and retardation of immunological inflammatory responses and its action may occur through the reduction of inflammatory mediators, including NO, $PGE_2$, and IL-6 production.

• The Korean Journal of Physiology and Pharmacology
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• v.13 no.4
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• pp.301-307
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• 2009

Inflammatory processes of vascular endothelial cells play a key role in the development ofatherosclerosis. We determined the anti-inflammatory effects and mechanisms of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) on LPS-treated human umbilical vein endothelial cells (HUVECs) to evaluate their cardioproteerive potential. Cells were pretreated with DHA, EPA, or troglitazone prior to activation with LPS. Expression of COX-2, prostaglandin $E_2$ ($PGE_2$) and IL-6 production, and $NF-{\kappa}B$ activity were measured by Western blot, ELISA, and luciferase activity, respectively. Results showed that EPA, DHA, or troglitazone significantly reduced COX-2 expression, $NF-{\kappa}B$ luciferase activity, and $PGE_2$ and IL-6 production in a dose-dependent fashion. Interestingly, low doses (10 ${\mu}$M) of DHA and EPA, but not troglitozone, significantly increased the activity of $NF-{\kappa}B$ in resting HUVECs. Our study suggests that while DHA, EPA, and troglitazone may be protective on HUVECs under inflammatory conditions in a dose-dependent manner. However there may be some negative effects when the concentrations are abnormally low, even in normal endothelium.

• Archives of Pharmacal Research
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• v.27 no.9
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• pp.923-929
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• 2004

Stylopine is a major component of the leaf of Chelidonium majus L. (Papaveraceae), which has been used for the removal of warts, papillomas and condylomas, as well as the treatment of liver disease, in oriental countries. Stylopine per se had no cytotoxic effect in unstimulated RAW 264.7 cells, but concentration-dependently reduced nitric oxide (NO), prostaglandin E$_2$ (PGE$_2$), tumor necrosis factor-a (TNF-$\alpha$) and interleukin-1$\beta$(IL-1$\beta$), and the IL-6 production and cyclooxygenase-2 (COX-2) activity caused by the LPS stimulation. The levels of inducible nitric oxide synthase (iNOS) and COX-2 protein expressions were markedly suppressed by stylopine in a concentration dependent manner. These results suggest that stylopine suppress the NO and PGE$_2$ production in macrophages by inhibiting the iNOS and COX-2 expressions. These biological activities of stylopine may contribute to the anti-inflammatory activity of Cheli-donium majus.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.23 no.2
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• pp.125-138
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• 2010

Objective : The present study was examined to evaluate the effects of Ecliptae Herba on the production of inflammatory mediators in vivo and in vitro. Methods : In cell viability, all three doses of Ecliptae Herba extract (25, 50 and $100\;{\mu}g/ml$) had no significant cytotoxicity during the experimental period. The increases of NO production and iNOS expression were detected in LPS-activated cells compared to control, but these increases were dose-dependently attenuated by pre-treatment with Ecliptae Herba extract. Results : 1. LPS plays a pivotal roles in inducing to the massive production of pro-inflammatory cytokines such as TNF-$\alpha$, IL-$1{\beta}$ and IL-6 in macrophages. 2. Ecliptae Herba extract reduced the elevated production of cytokines by LPS. 3. Ecliptae Herba extract reduced $PGE_2$ levels in a dose-dependent manner as a consequence of inhibition of COX-2 activity. 4. Ecliptae Herba extract significantly reduced the nuclear translocation of $NF-{\kappa}B$ induced by LPS. 5. In histopathological study, Ecliptae Herba effectively inhibited the increases of hind paw edema, skin thicknesses and inflammatory cell infiltrations induced by carrageenan treatment. Conclusions : These results provide evidences that therapeutic effect of Ecliptae Herba on the acute inflammation is partly due to the reduction of some of inflammatory factors by inhibiting iNOS and COX-2 through the suppression of $NF-{\kappa}B$.

• Journal of Korean Medicine Rehabilitation
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• v.24 no.4
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• pp.15-28
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• 2014

Objectives This study was carried out to find out the Antioxidant and Anti-inflammatory Effects of Components of Mahwangbujaseshin-tang in LPS-Stimulated RAW264.7 Macrophages. Methods There are 5 experimental groups. ; normal, control, EH (Ephedrae Herba), ALRP (Aconiti Lateralis Radix Preparata) and AR (Asiasari Radix). The extract of EH, ALRP and AR ($100{\mu}g/ml$) was added to each group. We examined cytotoxicity, total phenolic contents, DPPH and ABTS free radical scavenging activity, Intracellular ROS (reactive oxygen species) production, NO (Total Nitric oxide), iNOS (inducible nitric oxide synthase), PGE2 (prostaglandin E2), COX-2 (cyclooxygenase-2), $IL-1{\beta}$ ($interleukin-1{\beta}$), IL-6 (interleukin-6), $TNF-{\alpha}$ (tumor necrosis factor-${\alpha}$), MMP-9 (matrix metalloproteinase-9), TIMP-1 (tissue inhibitor of metalloproteinase-1) and HO-1 (heme oxygenase-1) expression level. Results 1. Total phenolic contents of EH were in the highest level. 2. DPPH and ABTS free radical scavenging activity of EH was in the highest level. 3. ROS production was significantly decreased in AR. 4. NO production was significantly decreased in EH, ALRP, AR and iNOS expression was decreased in EH, AR. 5. PGE2 and COX-2 expression was decreased in EH, AR. 6. $IL-1{\beta}$ production was significantly decreased in EH, AR and IL-6 production was significantly decreased in AR. $TNF-{\alpha}$ production was significantly decreased in ALRP, AR. 7. MMP-9 and TIMP-1 production were significantly decreased in EH. 8. HO-1 expression was significantly increased in EH. 9. With simultaneous usage of SnPP which is expression inhibitor of HO-1, NO, $IL-1{\beta}$, IL-6 and $TNF-{\alpha}$ production were partially increased in EH, ALRP, AR. Conclusions According to this study, Components of Mahwangbujaseshin-tang have anti-oxidants and anti-inflammation effects in LPS-Stimulated RAW264.7 Macrophages.

• Preventive Nutrition and Food Science
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• v.16 no.4
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• pp.333-338
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• 2011

In this study, antioxidative and anti-inflammatory activities of the herb (cinnamon, clove, glehnia root, ginger, violet-root cromwell, licorice, citrus peel and longan) extracts used for gamhongroju, one of the popular liqueurs in Korea, were investigated. Twenty grams of individual herbs were extracted in 60% purified ethanol and freeze-dried. A mixture of the individual herb extracts (HEM) was separately prepared. Cytotoxicity of the individual extracts and HEM on murine RAW264.7 macrophage cells were examined along with their recovering activity on $H_2O_2$-treated RAW264.7 cells. Antioxidant and anti-inflammatory activities of the extract-treated cells were determined by measuring superoxide dismutase (SOD) and glutathione peroxidase (GPx) activities, and Trolox equivalent antioxidant capacity (TEAC), nitric oxide (NO) and prostaglandin E2 (PGE2) levels. Violet-root cromwell extract showed the least cytotoxicity in terms of treated concentration. Most of the extracts, below levels of cytotoxicity, recovered the $H_2O_2$-treated cells. Treatment with some of the extracts increased SOD and GPx activities and TEAC levels while a majority inhibited the production of NO and PGE2 in lipopolysaccharide (LPS)-treated cells.

• Herbal Formula Science
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• v.20 no.2
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• pp.65-82
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• 2012

Objectives : The aim of this study was verification of the anti-oxidative and anti-inflammatory effects of Odukhwan(ODH) and Sasinhwan(SSH) in mouse macrophage, RAW264.7 cells. Methods : To investigate the anti-oxidative effect and scavenging activities of DPPH radical, superoxide anions, nitric oxide and peroxynitrite were measured. Cytotoxic activity of extract of ODH and SSH on RAW264.7 cells was measured using MTS assay. To proof the reductive activity of intracellular oxidation, DCFH-DA assay was performed. The nitric oxide(NO) production was measured and pro-inflammatory cytokines and $PGE_2$ were measured by ELISA kit. The levels of inducible nitric oxide synthase(iNOS), cyclooxygenase-2(COX-2) and nuclear NF-${\kappa}B$ p65 expression were detected by western blot. Results : After those analyses, we bring to a conclusion as follows. Both herbal formulations scavenged DPPH radical and nitric oxide. But ODH had no scavenging activity of superoxide anions and SSH had low scavenging activity in peroxynitrite. And the results indicated that ODH and SSH inhibited the LPS-induced NO, $PGE_2$ production and iNOS, COX-2 expression accompanied by an attenuation of IL-$1{\beta}$ and IL-6 production in RAW264.7 cells. They also have suppression effects of LPS-induced NF-${\kappa}B$ activation. Conclusions : ODH and SSH have anti-oxidative and anti-inflammatory effects and they may be a part of database for development of new anti-oxidative and anti-inflammatory drugs.

• The Korea Journal of Herbology
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• v.30 no.3
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• pp.35-40
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• 2015

Objectives : Zizyphus jujube (ZJ) has been used as a traditional medicine for various diseases. However, the inhibitory effect of ZJ on intestinal inflammation has not been fully understood, yet. The aim of this study is to investigate anti-colitis activity of ZJ in dextran sulfate sodium (DSS)-induced colitis mouse model. Methods : To investigate the protective effects of ZJ,the colitis mice were induced by drinking water containing 5% DSS for 7 days. Mice were randomized into groups receiving ZJ (500 mg/kg), sulfasalazine (SFZ) (150 mg/kg) as a positive control, or water as a negative control. We assayed the effects of ZJ on DSS-induced the clinical signs, measuring weight loss, colon length and disease activity index (DAI). Additionally, to find a possible explanation for the anti-inflammatory effects of ZJ, we evaluated the effects of ZJ on the production of prostaglandin $E_2$ ($PGE_2$) and expression of cyclooxygenase (COX)-2 in colitis tissue. Results : The results showed that mice treated with DSS showed considerable clinical signs, including weight loss, and reduced colon length. However, administration of ZJ significantly reduced the weight loss, shortens colon length, and improved DAI as clinical symptoms. Moreover, ZJ inhibited the $PGE_2$ production and COX-2 expression levels in DSS-treated colon tissues. Conclusions : Collectively, the findings of this study provide us with novel insights into the pharmacological actions of ZJ as a potential molecule for use in the treatment of intestinal inflammation including ulcerative colitis.