• 동의생리병리학회지
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• 제17권1호
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• pp.146-150
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• 2003

It had been known for a while that Crataegi Fructus(CF; Crataegus pinnatifida Bunge) had only a digestive effect. Recently, it has been demonstrated that CF also has an anti-hypertensive effect. However, its mechanism of relaxant effect has not been investigated yet. This study was examined to investigate the mechanism of vascular relaxation effect of CF in isolated rat thoracic aorta. CF revealed significant relaxation to phenylephrine(PE)-induced arterial contraction but much less to KCI-induced one. When CF was pretreated, it inhibited PE-induced contraction non-competitively. Methylene blue(10/sup -6/M) completely blocked the relaxant effect of CF whereas L-NAME(10/sup -5/M) did almost completely. However, atropine(10/sup -6/M) did not have any influence on vascular relaxation effect of CF. Regarding cNOS activity, CF significantly increased its activity from rat whole brain homogenate in a dose dependent manner which was inhibited by L-NAME(10/sup -5/M). On the other hand, CF did not affect on expression of TNF-α mRNA in RAW 264.7 cells, suggesting that CF is not related to iNOS activity. These results indicate that CF would be effective in relaxing vascular contraction through release of endothelial nitric oxide.

• Journal of Acupuncture Research
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• 제21권3호
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• pp.29-41
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• 2004

Objective : This study was produced to examine the effects of moxibustion that had been played important role to traditional oriental medical treatment on disease. Recently, it was reported that moxi-tar which is generated in the process of moxibustion as burning combustibles decreased NO and iNOS generation in C6-glioma and RAW 264.7 cells in our lab. Methods : C6-glioma cells were cultured in RPMI 1640 with FBS 10% in CO2 incubator. To study the protective effects of moxi-tar, we observed cell viability, DPPH activity, SOD activity, catalase activity and cell morphology after injury with $H_2O_2$. Results and Conclusions : Moxi-tar increased cell viability about twice as much as that of being injury by $H_2O_2$(moxi-tar $40{\mu}g/m{\ell}$, $H_2O_2$ $500{\mu}M$). And the results of free radical scavenger activity($80{\mu}g/m{\ell}$ : $78.91{\pm}4.4%$), SOD activity and catalase activity($80{\mu}g/m{\ell}$ : 21.6unit/mg protein) were increased by moxi-tar as dose-dependent manner. So we concluded that the effects of moxibustion which is played important role in Oriental medicine, might be free radical scavenger effects induced by moxi-tar. Conclusion : These results indicate that tBHP induces apoptosis through a lipid peroxidation-dependent mechanism and JS exerts the protective effect against the apoptosis by preventing peroxidation of membrane lipids.

• 한국식품영양학회지
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• 제28권3호
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• pp.343-350
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• 2015

Lonicera japonica 에탄올 추출물의 처리에 의하여 LPS에 의해서 활성화된 RAW 264.7 세포에서 NO의 생성량과 TNF-${\alpha}$, IL-$1{\beta}$ 및 IL-6와 같은 염증성 사이토카인의 분비를 억제하였고, MAPK family인 ERK, p38 및 JNK의 인산화와 $I{\kappa}-B{\alpha}$의 분해를 억제하였다. LPS로 유도한 endotoxin shock 동물실험에서 Lonicera japonica 에탄올 추출물 20 mg/kg에서 LPS로 유도한 endotoxin shock에 대한 생존율을 3배 이상 증가시켰으며, 생존시간도 1.3~1.4배 증가시켰다.

• 대한본초학회지
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• 제30권5호
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• pp.67-74
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• 2015

Objectives : TheHaedokgumhwa-sanwater extract (HDKHS) is used in Korea, Japan and China as a traditional therapeutic agent to cure an infectious disease. But its study is not enough. Therefore, the present study focused on the elucidation of HDKHS to investigate the anti-inflammatory effects and to established the possible mechanisms involved in its action on LPS-stimulated immune response in murine macrophages.Methods : Inflammatory status was induced by LPS and measured by increasement of inflammatory mediators. LPS induced secretions of NO and PGE₂in RAW 264.7 cells were measured using griess reagent and enzyme-linked immunosorbent assay (ELISA) kit respectively. production of IL-6 was examined using ELISA kit and expression of IL-6 mRNA was measured by RT-PCR method. To investigate the effects of HDKHS on inflammatory mediators, such as iNOS, COX-2 and MAPKs, western blot and RT-PCR were performed.Results : HDKHS significantly reduced production of NO and PGE2 which were induced by LPS. Also, activation of IL-6 was reduced both protein and mRNA levels. The expressions of inflammatory mediator include iNOS and COX-2 were decreased by pretreatment with HDKHS. futhermore The result showed HDKHS down-regulate the LPS induced phosphorylation of ERK 1/2, one of the MAPK family, which is considered as a main regulator of transmission from pathogens to nucleus of immune cells.Conclusions : Our results suggest that the anti-inflammatory properties of HDKHS may stem from the inhibition of pro-inflammatory mediators via suppression of initiation of inflammatory response by inhibiting MAPKs signaling pathways.

• Molecules and Cells
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• 제23권1호
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• pp.23-29
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• 2007

Cyclic AMP-responsive element binding protein (CREB) is known to be associated with angiogenesis. In the present study we investigated the possible role of CREB in the expression of vascular endothelial growth factor (VEGF) by mouse macrophages. Over-expression of CREB increased VEGF secretion by cells of the RAW264.7 mouse macrophage cell line. It also increased the promoter activity of a mouse reporter driven by the VEGF promoter, while a dominant negative CREB (DN-CREB) abrogated the activity, suggesting that CREB mediates VEGF transcription. Forskolin, an adenylyl cyclase activator, stimulated VEGF transcription, and the PKA inhibitor H89 abolished this effect. IFN-${\gamma}$, a potent cytokine, stimulated VEGF expression only in part through the PKA-CREB pathway. These results indicate that PKA phosphorylates CREB and so induces VEGF gene expression. An analysis of mutant promoters revealed that one of the putative CREB responsive elements (CREs), at -399 ~ -388 in the promoter, is critical for CREB-mediated VEGF promoter activity, and the significance of this CRE was confirmed by chromatin immunoprecipitation assays.

• Molecules and Cells
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• 제43권5호
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• pp.479-490
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• 2020

Interleukin-9 (IL-9) is well known for its role in allergic inflammation. For cancer, both pro- and anti-tumor effects of IL-9 were controversially reported, but the impact of IL-9 on tumor metastasis has not yet been clarified. In this study, IL-9 was expressed as a secretory form (sIL-9) and a membrane-bound form (mbIL-9) on B16F10 melanoma cells. The mbIL-9 was engineered as a chimeric protein with the transmembrane and cytoplasmic region of TNF-α. The effect of either mbIL-9 or sIL-9 expressing cells were analyzed on the metastasis capability of the cancer cells. After three weeks of tumor implantation into C57BL/6 mice through the tail vein, the number of tumor modules in lungs injected with IL-9 expressing B16F10 was 5-fold less than that of control groups. The percentages of CD4⁺ T cells, CD8⁺ T cells, NK cells, and M1 macrophages considerably increased in the lungs of the mice injected with IL-9 expressing cells. Among them, the M1 macrophage subset was the most significantly enhanced. Furthermore, peritoneal macrophages, which were stimulated with either sIL-9 or mbIL-9 expressing transfectant, exerted higher anti-tumor cytotoxicity compared with that of the mock control. The IL-9-stimulated peritoneal macrophages were highly polarized to M1 phenotype. Stimulation of RAW264.7 macrophages with sIL-9 or mbIL-9 expressing cells also significantly increased the cytotoxicity of those macrophages against wild-type B16F10 cells. These results clearly demonstrate that IL-9 can induce an anti-metastasis effect by enhancing the polarization and proliferation of M1 macrophages.

• Toxicological Research
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• 제16권1호
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• pp.1-8
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• 2000

Astrocytes generate free radicals including nitric oxide (NO) and reactive oxygen intermediates(ROI) which in turn play roles in the pathogenesis of degenerative diseases and sclerotic changes of the brain. This study was designed to evaluate the mechanism that free radicals contribute to the cytotoxicty of rat neonatal primary astrocytes. Treatment with NO donors alone including soldium nitroprusside(SNP), S-nitrosoglucathinoe (GSNO), and S-nitroso-n-acetylpenicillamine (SNAP) showed a little effect on the death of rat neonatal primary astrocytes, whereas SNP markedly induced the death of RAW 264.7 cells. ROI inculding H2O2 and O2 donor also slightly induced the death of rat primary astrocytes. However, 3-morpholinosydnonimine(SIN-1), a donor of peroxynitrite (ONOO), which is a reactive compound of NO with superoxide, significantly decreased the viability of rat primary astrocytes in a dose-dependent manner. Cells were retarded in outgrowth of viability of cellular processes with cell shrinkage and detachment from culture dishes. Hoechst staining demonstrated that SIN-1-induced cell death might be due to an apoptosis which was characterized by nuclear condensation and fragmentation. SIN-1-induced apoptosis was prevented by the pretreatment with superoxide dismutase (SOD) and catalase in rat primary astorocytes. Furthermore, prevention of the generation of reduced glutathione (GSH) by DL-buthionine-[S, R]-sulfoximine (BSO) aggravated the cytotoxic effects of SNP, benzene triol, and SIN-1 in rat primary astrocytes. Taken together, it is suggested that peroxynitrite may be a major effector of apoptosis and cellular antioxidant system is important for cell survival in rat prima교 astrocytes.

• 생약학회지
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• 제39권4호
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• pp.295-299
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• 2008

The antioxidant activities, anti-inflammatory activity and cytotoxic effects of methanol extract from Euphorbia humifusa were evaluated in this study. Total phenolic compound contents were $68.35{\pm}0.16$ mg/g and total flavonoid compound contents were estimated as $38.74{\pm}1.26$ mg/g. EC50 values for DPPH radical scavenging activity of methanol extract was $56.26{\pm}0.66{\mu}g/mL$ and those of positive controls as ascorbic acid, ${\alpha}$-tocopherol and BHA were $8.38{\pm}0.14{\mu}g/mL$, $16.45{\pm}0.89{\mu}g/mL$ and $21.18{\pm}1.01{\mu}g/mL$ respectively. NO scavenging activity increased in depending on concentration of extract. Treatment of RAW 264.7 cells with extract caused inhibition of LPS-induced nitric oxide production. The cell viability showed that the methanol extract had cytotoxicity in the growth of breast cancer cell ($66.54{\pm}1.91%$ at $400{\mu}g/mL$ conc., $43.98{\pm}3.35%$ at $800{\mu}g/mL$ conc.). Based on the results, It was suggested that the methanol extract of Euphorbia humifusa has a potential candidate for functional cosmetic and medicine.

• 한방재활의학과학회지
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• 제24권4호
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• pp.1-13
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• 2014

Objectives This study was to investigate the antioxidative capacity, antiobesity effect and anti-diabetes effects of Mori Fructus and dried Mori Fructus in Raw 264.7 cells and 3T3-L1 cells. Methods 3 different types of Mori Fructus extracts (water 100%, ethanol 30%, ethanol 100%) were used in this study. And 3 different types of dried Mori Fructus extracts (water 100%, ethanol 30%, ethanol 100%) were used in this study. Total polyphenol compund, total favonoid compound, DPPH radical scavenging, ROS activity, NO, cell proliferation were measured in the experiment. Expressions of adipogenic transcription factors including $C/EBP-{\alpha}$, $C/EBP-{\beta}$, $PPAR-{\alpha}$, $PPAR-{\gamma}$, $AMPK-{\alpha}$ were analyzed by Real time PCR. Results Mori Fructus extracts measurements are higher than dried Mori Fructus extracts measurements at Total flavonoid compound and total flavonoid compound. Mori Fructus extracts measurements are lower than dried Mori Fructus extracts measurements at DPPH radical scavenging, ROS activity, NO. In RT-PCR analysis, there is a tendency that dried Mori Fructus extracts inhibit the expression of $C/EBP-{\alpha}$, $C/EBP-{\beta}$ genes. In RT-PCR analysis, there is a tendency that dried Mori Fructus extracts promote the expression of $PPAR-{\alpha}$, $PPAR-{\gamma}$, $AMPK-{\alpha}$ genes. Conclusions Mori Fructus is effective on inhibiting the oxidation and dried Mori Fructus is effective on inhibiting the obesity and diabetes.

• 동의생리병리학회지
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• 제20권2호
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• pp.436-441
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• 2006

Rheumatoid arthritis is a chronic, systemic, and inflammatory autoimmune disorder that affects 1% of the adult population worldwide. Osteoarthritis is a multifactorial disease with high morbidity that is characterized by degradation of the matrix and destruction of articular cartilage. In this study, we examined the inhibition effect of Trachelospermi Caulis on the inflammation($TNF-{\alpha}$, $IL-1{\beta}$, NO), cartilage protection(MMP-13), and cell death in arthritis. RAW 264.7 and SW 1353 cells were cultivated in DMAE(GibcoBRL, USA) with 5% FBS and Fungizone in $37^{\circ}C$, 5% CO2. THP-1 cells were cultivated in RPMI(GibcoBRL, USA) with 5% FBS and Fungizone in $37^{\circ}C$, 5% CO2. Activity of caspase-3, XIAP, Cytochrome C in the cell was examined by using western blot. The results obtained were as Follows; Concentration of nitric oxide in Trachelospermi Caulis treatment group significantly decreased compared with that of non-treatment group (P<0.05). In treated group, Concentration of Trachelospermi Caulis was not significantly associated with cell death. Concentration of $TNF-{\alpha}$ and $IL-1{\beta}$ in Trachelospermi Caulis treatment group decreased significantly compared with that of none treatment group (P<0.05). Relative density of MMP-13 in Trachelospermi Caulis treatment group decreased significantly compared with that of none treatment group and dose-response relationship was observed. After treatment of staurosporin in SW1353 which increases cell death, in Trachelospermi Caulis treated group, the cell death was effectively decreased. In conclusion, these results suggest that Trachelospermi Caulis inhibit inflammation and cell death in arthritis. More researches about effect of Trachelospermi Caulis are considered to need.