• 동의생리병리학회지
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• 제20권4호
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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.

• 동의생리병리학회지
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• 제20권3호
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• pp.690-696
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• 2006

Houttuynia cordata Thunb, well known as 'E-Sung-Cho' in Korea, is a traditional medicinal plant generally used in Oriental medicine therapy. In the present study, we investigated the effect of water extract of H. cordafa (WEHC) on the growth of human breast carcinoma MCF-7 cells. Exposure of MCF-7 cells to WEHC resulted in growth inhibition and apoptosis in a dose-dependent manner as measured by MTT assay and fluorescent microscope. The anti-proliferative effect of WEHC was associated with a dose-dependent up-regulation of cyclin-dependent kinase inhibitor p21 in a p53-independent fashion. We found WEHC 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 $E_2{\;}(PGE_2)$ synthesis. Taken together, these findings provide important new insights into the possible molecular mechanisms of the anti-cancer activity of H. cordata.

• Biomolecules & Therapeutics
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• 제16권3호
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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.

• 약학회지
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• 제48권6호
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• pp.317-322
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• 2004

The use of nonsteroidal anti-inflammatory drugs (NSAIDs) is limited by their ability to induce gastrointestinal injury. It has been shown that nitric oxide (NO), similar to pro staglandins (PGs), appears to play an important role in gastric mucosal defence. We hypothesized that NSAIDs contained NO group would be less acutely toxic to the gastric mucosa, but would not interfere with their ability to suppress inflammatory process in rats. We have compared the ulcerogenic and anti-inflammatory effect of CW-501029 (NO-NSAIDs), CW-501027 (NSAIDs) and indomethacin. Both did not change mean blood pressure and heart rates, indicating that they had no side effect on cardiovascular system. We found that CW-501029 increased nitrite/nitrate levels without changing of blood pressure and heart rates. We suggest that it may help gastric mucosal blood flow, the which helps reducing the discomfort in astrointestinal system. Carrageenan-induced PGE2 increase was reduced in a similar tendency when compared CW-501027 or CW-501027 with control in back exudate of rats, but CW-501029 less reduced PGE2 than CW-502027 or indomethacin in gastric tissues. CW-501027 or CW-501029 reduced platelet aggregation. From these results we suggest that CW-501029 may improve the side effect by reduction of short-term gastric injury and less inhibition of PGs synthesis.

• 동의생리병리학회지
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• 제22권5호
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• pp.1315-1321
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• 2008

본 논문은 오랫동안 민간요법으로 염증치료에 사용되어오던 섬수가 lipopolysaccharide(LPS)-자극된 BV2 소교 세포의 nitric oxide(NO) 분비에 미치는 효과에 대해 연구한 내용이다. 실험 결과 섬수는 세포 생존력에 대한 영향 없이 BV2 소교 세포에서 NO 분비를 억제시켰고, nitric oxide synthase (iNOS) 단백질도 감소시켰다. 또한 섬수는 prostaglandin E2 (PGE2) 생산 및 cyclooxygenase (COX)-2 발현을 저지하였고, proinflammatory cytokines과 ${IkB-\alpha}$감소를 억제시켰다. 따라서 섬수가 $I{\kappa}B-{\alpha}$감소를 억제함으로써 NO 합성을 저해하여 항염증작용을 할 수 있다는 내용이다.

• Journal of Ginseng Research
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• 제37권1호
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• pp.54-63
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• 2013

Ginsenoside Rd is a primary constituent of the ginseng rhizome and has been shown to participate in the regulation of diabetes and in tumor formation. Reports also show that ginsenoside Rd exerts anti-oxidative effects by activating anti-oxidant enzymes. Treatment with ginsenoside Rd decreased nitric oxide and prostaglandin $E_2$ ($PGE_2$) in lipopolysaccharides (LPS)-challenged RAW264.7 cells and in ICR mouse livers (5 mg/kg LPS; LPS + ginsenoside Rd [2, 10, and 50 mg/kg]). Furthermore, these decreases were associated with the down-regulations of inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX)-2 and of nuclear factor (NF)-${\kappa}B$ activity in vitro and in vivo. Our results indicate that ginsenoside Rd treatment decreases; 1) nitric oxide production (40% inhibition); 2) $PGE_2$ synthesis (69% to 93% inhibition); 3) NF-${\kappa}B$ activity; and 4) the NF-${\kappa}B$-regulated expressions of iNOS and COX-2. Taken together, our results suggest that the anti-inflammatory effects of ginsenoside Rd are due to the down-regulation of NF-${\kappa}B$ and the consequent expressional suppressions of iNOS and COX-2.

• Tuberculosis and Respiratory Diseases
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• 제70권2호
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• pp.113-124
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• 2011

Background: Macrophages are one of the most important inflammatory cells in innate immunity. PU.1 is a macrophage-specific transcription factor. Ubiquitins are the ultimate regulator of eukaryotic transcription. The ubiquitination process for PU.1 is unknown. This study investigated the lipopolysaccharide (LPS)-induced activation of PU.1 and its relation to ubiquitins in the macrophages. Methods: Raw264.7 cells, the primary cultured alveolar, pulmonary, and bone marrow derived macrophages were used. The Raw264.7 cells were treated with MG-132, $NH_4Cl$, lactacytin and LPS. Nitric oxide and prostaglandin D2 and E2 were measured. Immunoprecipitation and Western blots were used to check ubiquitination of PU.1. Results: The PU.1 ubiquitination increased after LPS ($1{\mu}g$/mL) treatment for 4 hours on Raw264.7 cells. The ubiquitination of PU.1 by LPS was increased by MG-132 or $NH_4Cl$ pretreatment. Two hours of LPS treatment on macrophages, PU.1 activation was not induced nor increased with the inhibition of proteasomes and/or lysosomes. The ubiquitination of PU.1 was increased in LPS-treated Raw264.7 cells at 12- and at 24 hours. LPS-treated cells increased nitric oxide production, which was diminished by MG-132 or $NH_4Cl$. LPS increased the production of $PGE_2$ in the alveolar and peritoneal macrophages of wild type mice; however, $PGE_2$ was blocked or diminished in Rac2 null mice. Pretreatment of lactacystin increased $PGE_2$, however it decreased the $PGD_2$ level in the macrophages derived from the bone marrow of B57/BL6 mice. Conclusion: LPS treatment in the macrophages ubiquitinates PU.1. Ubiquitination of PU.1 may be involved in synthesis of nitric oxide and prostaglandins.

• 대한한의학회지
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• 제29권1호
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• pp.156-166
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• 2008

Objective : Pamooltang (PM) and Sipjeondaebotang (SC) are used in Korea to treat many diseases such as sterility, menstrual disorder, and general prostration. We made a comparative study of PM and SC which are different in component ratio between Astragalus membranaceus BUNGE (AC) and Cinnamomum cassia PRESL. (CC). Methods : Anti-oxidation was studied by 1.1.-diphenyl-2-picrylhydrazyl (DPPH) assay and anti-inflammation was investigated by prostaglandin $E_2\;(PGE_2)$ and nitric oxide (NO) inhibition assay. For immune response activities, this study used NO synthesis on RAW 264.7 cells and splenocyte proliferation. Results : The results showed that PM and SC components had no significant effect of anti-oxidation or anti-inflammation. However, we observed their effects upon inducible NO synthesis in Raw 264.7 cells. The SC2 stimulated NO synthesis $11.42\pm1.36{\mu}M$ (control; $0.89\pm0.00{\mu}M$). PM and SC components had the effect of immune response which in a dose-dependent manner significantly induced the splenocyte proliferation. The splenocyte proliferation induced by SC2 was higher than others at the concentration of 1, 10, 100, 1000 and 2000 ${\mu}g/ml$. The SC8 was shown to up-regulate IgG, 100 ${\mu}g/ml$ 3.3 times, 1000 ${\mu}g/ml$ 2.6 times as a control. Conclusions : These results may have important implications for our understanding of the ratios of AC and CC in SC.

• 한국식품영양과학회지
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• 제41권7호
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• pp.907-913
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• 2012

포도, 대황, 사탕수수 등을 포함한 다양한 식물에서 발견되는 hydroxystilbene의 일종인 piceatannol은 암세포의 증식을 억제하고 apoptosis를 유발하는 것으로 알려져 있다. 본 연구에서는 AGS 인체위암세포를 대상으로 piceatannol에 의한 암세포 증식억제 과정에서 나타나는 또 다른 현상들을 조사하기 위하여 실시되었다. Piceatannol이 처리된 AGS 위암세포는 piceatannol의 처리 농도의 증가에 따라 생존율이 감소되었으며, 이는 세포주기 G1 arrest 유발과 연관이 있음을 MTT assay와 flow cytometry 분석을 통하여 확인하였다. Piceatannol에 의한 AGS 세포의 G1 arrest는 Cdks 및 cyclins의 발현 변화 및 Cdk 저해제인 p21의 발현을 전사 및 번역 수준에서 증가시켰으며, pRB 단백질의 인산화 감소 및 E2F1의 발현 억제와 연관성이 있었다. 아울러 piceatannol은 COX-2의 mRNA 및 단백질의 발현을 억제하였으나 COX-1의 발현에는 영향을 미치지 않았으며, piceatannol에 의한 COX-2의 발현억제는 PGE2의 생성 저하와 관련이 있었다. 본 연구의 결과는 piceatannol에 의한 세포주기 G1 arrest 유발이 COX-2의 선택적 발현 차단과 연관이 있음을 보여 주는 것이다.

• 대한화장품학회지
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• 제45권2호
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• pp.117-129
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• 2019

올벚나무 잎 추출물을 제조하고 LPS로 자극시킨 RAW 264.7 대식세포에 대한 항염 효과를 조사하였다. 추출물 및 분획물(n-hexane, ethyl acetate, n-butanol 및 water)에 대하여 활성 실험을 진행한 결과, $100{\mu}g/mL$ 농도에서 ethyl acetate (EtOAc) 분획물이 세포독성 없이 76.3%의 NO 생성 억제 효과를 나타내었다. 또한 EtOAc 분획물이 농도 의존적으로 NO 생성을 효과적으로 억제하고 있음을 확인하였다. 이러한 항염 효과에 대한 추가적인 기전 연구를 진행한 결과, EtOAc 분획물이 $PGE_2$, $IL-1{\beta}$ 및 IL-6의 생성을 감소시켰으며, iNOS와 COX-2 단백질의 발현도 억제시키는 것을 확인하였다. EtOAc분획물에서 활성 성분을 규명하기 위하여 column chromatography를 진행하였으며 5개의 화합물을 분리 동정하였다. ursolic acid (1), prunasin (2), methyl p-coumarate (3), kaempferol (4), astragalin (5). 분리된 compound 1 - 5는 모두 올벚나무 잎에서 처음 분리된 물질이다. 분리된 화합물을 대상으로 활성 실험을 진행한 결과, 플라보노이드 compound 4와 5가 NO 생성을 억제하고 있음을 확인하였다. 이상의 결과를 바탕으로, 플라보노이드를 유효성분으로 함유하는 올벚나무 잎은 항염 효과를 갖는 천연 소재로서 활용이 가능할 것이라 사료된다.