• 한국식품과학회지
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• 제28권5호
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• pp.850-858
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• 1996

본 연구는 녹차를 열수로 추출하여 LDL에 대한 항산화 활성을 연구한 결과이다. 녹차 1.25 g에 열수 500 ml를 가하여 추출한 후 다시 500 ml를 가하여 재추출하였으며 이를 증발, 건조한 결과 추출액의 건물량은 4.67 mg이었다. 녹차잎의 함량이 1.25%가 되도록 조절한 녹차에는 항산화 활성이 강한 polyphenol 화합물중 (-) epicatechin gallate 54.1%, (-) epicatechin gallate 26.2%, (-) epigallocatechin 10.7%, (-) epicatechin 이 7.0% 및 catechin이 1.8% 함유되었다. Low density lipoprotein (LDL)의 산화에 있어서 LDL은 $CuSO_4$ 존재하에서 macrophage와 배양시켰더니 빠르게 산화를 일으켰다. 그러나 $5\;{\mu}M\;CuSO_4$ 매개 LDL의 산화에 50 및 $100\;{\mu}g/ml$ 농도의 GTE를 넣어 배양하면 거의 완전히 산화를 억제하였다. $5\;{\mu}M\;CuSO_{4}$ 존재하에서 산화시킨 LDL의 전기영동 이동거리는 native LDL보다 높았다. 또 50 및 $100\;{\mu}g/ml$ 농도의 GTE는 J774, human monocyte derived macrophags 및 vascular endothelial cells에서 유도되는 LDL의 산화를 억제하였다. $CuSO_4$ 및 cell 유도에 의하여 산화되는 LDL은 native LDL보다 더욱 많은 양이 human macrophage에 의하여 분해되고 GTE는 macrophage에 의한 $^{125}I-LDL$의 산화를 강력히 억제하였다. 그리고 GTE는 cell 매개 LDL의 macrophage에 의한 축적 또는 분해를 억제하였다. LDL을 $5\;{\mu}M\;CuSO_4$, 존재하에서 산화시킬 때 GTE를 50 및$100\;{\mu}g/ml$ 농도로 넣어 배양하면 공액 dienes의 생성이 억제되고, 또한 Oxid LDL macrophage derived human monocytes에서도 축적이 억제되었다.

• Journal of Nutrition and Health
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• 제42권5호
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• pp.434-441
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• 2009

본 연구는 J774 대식세포에서 FPN 유전자 발현 조절에 구리가 미치는 영향을 알아보기 위하여 수행되었으며 그 결과는 다음과 같다. J774 대식 세포에 구리를 처리하였을 때, iron exporter FPN의 mRNA 수준이 농도 의존적으로 증가하는 것으로 나타났다. 반면, iron importer DMT1의 mRNA 수준은 구리 처리에 의해 영향을 받지 않았다. Actinomycin D를 이용하여 mRNA 합성을 억제한 상태에서 FPN mRNA 분해 정도를 시간별로 추적한 결과, acitnomycin D 처리 후 9시간 경과시 FPN mRNA 수준이 처음 수준의 약 60% 정도로 감소하였다. 배양액에 구리를 첨가한 경우에도 FPN mRNA의 분해 정도는 아무것도 처리하지 않은 대조군과 유의적인 차이가 없었으며, 이로 볼 때 구리는 FPN mRNA의 안정성에 영향을 미치지 않는 것으로 생각된다. 한편, reporter assay 실험 결과 구리의 첨가는 FPN 프로모터 활성을 유의적으로 증가시키는 것으로 나타나, 구리가 FPN mRNA의 전사 과정을 직접적으로 촉진함을 알 수 있었다. 또한, FPN 5'-UTR에 위치하는 IRE (iron response element)의 존재 여부는 구리에 의한 FPN 전사 개시 활성에 영향을 주지 않는 것으로 나타났으며, 이로 볼 때 구리는 철분과는 독립적인 작용 기작에 의해 FPN 유전자 발현을 조절하는 것으로 사료된다. 이상의 결과를 종합해 볼 때, 구리는 대식 세포에서 전사개시 과정을 활성화함으로써 농도 의존적으로 FPN 유전자 발현을 촉진하는 것으로 생각되며, 이는 구리가 철분의 대사에 미치는 새로운 작용 기작을 제시한다. 앞으로, 구리와 철 분의 상호 작용이 FPN의 철분 및 다른 무기질 이온의 세포내 외 수송 (transport)에 어떤 영향을 미치는지에 대한 기능적 연구가 계속적으로 이루어져야 할 것으로 사료된다.

• Journal of Microbiology and Biotechnology
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• 제16권1호
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• pp.136-140
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• 2006

In this study, we investigated the synergistic effects of Staphylococcus aureus extracts (membranes and walls) and lipopolysaccharide (LPS) derived from Escherichia coli on tumor necrosis factor (TNF) production in the pathogenesis of silica-induced inflammation. The synergistic induction of TNF by silica particles $(<20\;{\mu}m)$ in combination with either S. aureus extracts or LPS was examined in J774A.1 cell cultures. Media from the treated and untreated cell cultures were assayed for TNF, using the mouse WEHI 164 cell cytotoxicity assay and enzyme immunoassay. Cells exposed simultaneously to silica and $0.5\;{\mu}g/ml$ S. aureus extracts (or 0.5 ng/ml of LPS) produced a significantly higher level of TNF than those produced by the inducer alone. Our results indicate that device-associated infections (or pyrogen contamination) could enhance inflammatory responses, because of particles produced by the wear of medical implants or particulate biomaterials used for clinical purposes.

• Parasites, Hosts and Diseases
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• 제53권1호
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• pp.21-27
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• 2015

Plants used for traditional medicine contain a wide range of substances that can be used to treat various diseases such as infectious diseases. The present study was designed to evaluate the antileishmanial effects of the essential oil and methanolic extract of Myrtus communis against Leishmania tropica on an in vitro model. Antileishmanial effects of essential oil and methanolic extract of M. communis on promastigote forms and their cytotoxic activities against J774 cells were evaluated using MTT assay for 72 hr. In addition, their leishmanicidal activity against amastigote forms was determined in a macrophage model, for 72 hr. Findings showed that the main components of essential oil were ${\alpha}$-pinene (24.7%), 1,8-cineole (19.6%), and linalool (12.6%). Findings demonstrated that M. communis, particularly its essential oil, significantly (P<0.05) inhibited the growth rate of promastigote and amastigote forms of L. tropica based on a dose-dependent response. The $IC_{50}$ values for essential oil and methanolic extract was 8.4 and $28.9{\mu}g/ml$ against promastigotes, respectively. These values were 11.6 and $40.8{\mu}g/ml$ against amastigote forms, respectively. Glucantime as control drug also revealed $IC_{50}$ values of 88.3 and $44.6{\mu}g/ml$ for promastigotes and amastigotes of L. tropica, respectively. The in vitro assay demonstrated no significant cytotoxicity in J774 cells. However, essential oil indicated a more cytotoxic effect as compared with the methanolic extract of M. communis. The findings of the present study demonstrated that M. communis might be a natural source for production of a new leishmanicidal agent.

• Natural Product Sciences
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• 제27권4호
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• pp.228-233
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• 2021

Bioactivity-guided fractionation by preliminary screening using interleukin-1β production in lipopolysaccharides (LPS)-induced J774A.1 cell line led to the isolation of fourteen structures including chromone, isocoumarins, flavanoids, and triterpenes from the aerial part of Agrimonia pilosa Ledeb. All structures were determined by measuring their spectroscopic data and comparing their spectroscopic data with the literatures. All the isolates were tested for their inhibitory activities against interleukin-1β production in LPS-induced J774A.1 cell. Of the tested compounds, (S)-(+)-5,7-dihydroxy-2-(1-methylpropyl)chromone (1), agrimonolide-6-O-β-D-glucopyranoside (5), agrimonolide-6-O-α-L-arabinofuranosyl-(1→6)-β-D-glucopyranoside (6), and catechin (10) were found to be active. Furthermore, compound 1 suppressed the protein expressions of NLRP3 and NLRC4 in murine macrophage.

• 한국식품영양과학회지
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• 제29권2호
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• pp.312-315
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• 2000

본 연구에서는 우리밀로부터 대식세포 식작용 활성화시키는 물질을 탐색하는 과정에서 고분밀의 EA 추출물이 가장 높은 식작용 활성 증강효과를 보임을 알았고, EA 추출물을 정성한 후 순수 분리 정제하여 대식세포 활성 증강효과를 나타내는 단일 물질을 찾고자 하였다. TLC 방법을 이용하여 ASW와 고분밀의 EA 추출물을 정성한 결과 약 5개 정도의 band를 확인한 후, column chromatography를 이용하여 EA 추출물을 분획하였다. 1차 column chromatography만으로는 single fractionation이 어려워 분획물을 fraction A, B, C, D로 분류하여 식작용 활성 증강효과를 측정하였다. 대분류된 각 fraction의 식작용 활성 증강효과를 측정한 결과 고분밀의 fraction b가 대부분의 활성을 가진 분획으로 사료되었다. 2차 column chromatography를 통해 fraction b만응 분리 하였으며, 구조결정 실험과 동물실험을 추가로 시행하여 fraction b의 효과를 입증할 것이다.

• Journal of Life Science
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• 제9권2호
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• pp.57-61
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• 1999

This study was designed to develope the antioxidative activity on oxidation of human low density lipoprotein(LDL) from marine microbials. Bacillus KS-96 producign antioxidant have been isolated and identified from seawater, Bacillus sp. KS-96. The optimal medium pH was 7.0 and incubation temperature was 30$^{\circ}C$. The antiosidant of potential substance produced extracellularly in the culture broth by Bacillus sp. KS-96 was obtained by elution of silica gel culumn chromatography with hexane, ethylacetate and water. The ethylacetate faction are shown at highest level of antioxidant activity using thiocyanate method among them. By IR, NMR, and GC/MS, antioxidant purified from ehtylacetate fraction was identified and named as 2,6-dimethoxyphenol. 2,6-dimethoxyphenol inhibited the metal mediated oxidation of human LDL at concentration of 50∼100 ${\mu}$g/mL in the presence of 5uM CuSO4 with macrophage or J774 cells.

• Biotechnology and Bioprocess Engineering:BBE
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• 제5권5호
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• pp.366-371
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• 2000

The aim of this study was to investigate the efficiency of the pentagalloic acid compound in inhibiting the metal ions and cell lines that mediate in low density lipoprotein (LDL) oxidation. Pentagalloic acid prolonged the lag time preceeding the onset of conjugated diene formation. In chemically induced LDL oxidation by Cu$^2$(sup)+ plus hydrogen peroxide or peroxyl radical generated by 2, 2-azo-vis (2-amidino propane) hydrochloride (AAPH), pentagalloic acid inhibited LDL oxidation as monitored by measuring the thiobarbituric acid reactive substances(TBARS), malondialdehyde(MDA), and gel electrophoretic mobility. The physiological relevance of the antioxidative activity was validated at the cellular level where pentagalloic acid inhibited mouse macrophage J774 and endothelial cell-mediated LDL oxidation. When compared with several other antioxidants, pentagalloic acid showed a much higher ability than naturally occuring antioxidants, ${\alpha}$-tocopherol and ascorbic acid, and the synthetic antioxidant, probucol.

• 한국미생물·생명공학회지
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• 제25권3호
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• pp.286-292
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• 1997

This study was undertaken to evaluate an antioxidative activity of silymarin and silybin obtained from Silybum marianum against oxidation of human low density lipoprotein (LDL). The electrophoretic mobility observed apparently was higher phase for LDL oxidized by macrophages compared to native LDL. Silymarin and silybin inhibited the copper-catalysed oxidation of human LDL in a dose-dependent manner. Silymarin and silybin at the concentration of 50 $\mu$M/ml also inhibited the copper catalysed oxidation of LDL induced by the cell J774 and macrophages. LDL reisolated from the cell incubation in the presence of silymarin or silybin was degraded at rates similar with native LDL. Silymarin or silybin found to be potential inhibitors against oxidation of $^{125}$I-LDL by macrophages and endothelial cells.

• 대한한방내과학회지
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• 제26권3호
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• pp.575-586
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• 2005

FC(free choresterol) plays an important role in normal and pathophysiological cells including that of messenger molecule or dilator of blood vessels in such illnesses as artheriosclerosis, hypertension and myocardial infarction. Smooth muscle and endothelial cell functions in the arteria wall are unified by complex intercellular signalling processes. In arteria comprised of one layer of smooth muscle cells surrounding the endothelium, the close apposition of the two cell types enables a signal derived from one cell to rapidly diffuse to neighboring cells. Experimentation was conducted to investigate the potential contribution of Sohaphyangwon(SHHW) on levels of FC generated by goaded microphages, and mechanisms of protection against ACAT inhibitor. It was found that J774 macrophages, which normally do not express FC were expressed by oxLDL and ACAT inhibitor. SHHW protected cells were found to be resistant to oxLDL and delayed death following the FC. Inhibition of FC formation abolished the protective effect against ACAT inhibitor exposure. Cadiovascular diseases include abnormalities of blood vessels dysfunction of the renin-angiotension system. What relation herbal medicine may have with vessel endothelium necrosis was here studied. In Oriental Medicine, SHHW water extract used for diseases in relation to cardiovascular systems. The resistence to cardiovascular disease of ACAT inhibitor induced J774 macrophage cells were studied through analysis of cell morphological patterns and immunochemistry of SHHW. The results of this study suggest that SHHW has protective effects on the cardiovascular system, and that it is effective in both prevention and treatment of diseases of the cardiovascular system, particularity against necrosis of blood.