• Journal of Applied Biological Chemistry
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• 제66권
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• pp.519-526
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• 2023

본 연구는 마우스 대식세포 유래 Raw264.7 세포주에서 황기와 산초 1:1 혼합물(AZM-1:1)의 면역 증진 효능을 탐색하였다. Raw264.7 세포에 100-400 ㎍/mL의 A ZM-1:1 처치는 세포 생존율의 변화 없이 inducible nitric oxide synthase mRNA의 발현 증가와 함께 nitric oxide의 생성을 통계적으로 유의하게 증가시켰다. 더불어 A ZM-1:1은 처치 농도 의존적으로 cyclooxygenase-2 mRNA의 유도와 함께 세포 배양액 중 prostaglandin E₂의 함량을 증가시켰다. 또한, AZM-1:1은 tumor necrosis factor-α, interleukin-1β, interleukin-6 및 monocyte chemoattractant protein-1의 전사를 촉진하였다. Immunoblot 분석을 통하여 AZM-1:1은 mitogen-activated protein kinase의 인산화를 증가시키고, inhibitory-κBα의 인산화를 매개한 분해를 촉진하며, p65의 인산화를 증가시킬 수 있음을 확인하였다. AZM-1:1의 처치는 녹색 형광으로 표지된 대장균 파편의 탐식작용을 촉진하였다. 따라서, 이상의 결과는 A ZM-1:1가 대식세포를 포함한 내재면역을 증진시키는 기능성 식의약 소재가 될 수 있음을 나타낸다.

• 한국응용과학기술학회지
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• 제33권3호
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• pp.606-613
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• 2016

본 연구는 커피부산물의 화장품 소재로서의 활용가능성을 평가하기 위해 커피부산물을 유기용매인 n-hexane을 이용하여 $60({\pm}10)^{\circ}C$에서 24시간동안 교반하며 추출하여 실험을 수행하였다. B16F10 melanoma cell line과 RAW264.7 macrophage cell line에서의 커피박 추출물의 세포독성을 water solubletetrazolium salt-1 assay로 평가하였다. 또한 free radical 소거능을 측정하기 위해 DPPH법을 사용하여 항산화를 평가하였으며, 항균력 검색을 위해 Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli, Candida albicans를 이용하였다. 실험결과 B16F10 murine melanoma cells에서 커피부산물 추출물을 처리한 군은 $0.125{\sim}2{\mu}{\ell}/m{\ell}$ 농도에서, RAW 264.7 macrophage cells에서는 0.125부터 $0.5{\mu}{\ell}/m{\ell}$ 농도에서 세포독성을 나타내지 않았다. 항산화 실험 결과 커피박 추출물은 농도 의존적인 DPPH radical 소거능을 보였다. 또한 커피박 추출물의 항균 효능을 측정하기 위해 Paper disc법을 이용하였으며 그 결과 Straphylococcus epidermidis, Straphylococcus aureus, Escherichia coli, Candida albicans에서 각각 $11.3{\pm}0.4$, $12.{\pm}0.7$, $12.0{\pm}0.0$, $0.0{\pm}0.0mm$의 clear zone을 나타내었다. 이러한 결과는 커피박 추출물의 화장품 소재로서의 가치를 제안할 수 있다.

• 한국식품저장유통학회지
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• 제30권1호
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• pp.65-77
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• 2023

In this study, we investigated the immunological properties of six strains of Acetobacter pasteurianus through nuclear factor-kappa B/activator protein-1 (NF-κB/AP-1) transcription factor activation and nitric oxide (NO) and cytokine production in macrophages. We found that the six A. pasteurianus strains had no significant inhibitory effect on the cell viability of RAW-Blue^TM cells at the concentration of (25, 50, 100 CFU/macrophage). The production of NO and cytokines (TNF-α, IL-1β, and IL-6) showed different abilities of immune activation for each strain, and it was 0.7 to 0.9 times higher than that of the LPS (100 ng/mL, v/v) positive control and 7 to 8 times superior to that of the negative control group. To explore the underlying mechanism, we evaluated the mRNA expression of pro-inflammatory genes. Consequently, we found that inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX)-2 expression including genes expression of cytokines were elevated by the six A. pasteurianus treatment. These results suggested that the six strains of A. pasteurianus have an excellent industrial application value as a functional material for the purpose of enhancing immune function.

• 한국식품과학회지
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• 제34권4호
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• pp.732-736
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• 2002

더덕열수추출물의 면역증강효과를 조사하였다. 더덕열수추출물을 1에서 $25\;{\mu}g/mL$의 농도로 첨가했을 때 BSA로 면역된 마우스의 림파절 세포는 무첨가군에 비해 2.8에서 11.2배의 세포증식 효과를 나타냈다. 더덕열수추출물은 인삼열수추출물보다 증식효과가 높게 나타났으며, 면역증강효과가 알려진 B. adolescentis M101-4 와는 유사한 정도의 효과를 보였다. 비장 및 Peyer's patch 세포에 대해서는 1에서 $25\;{\mu}g/mL$의 더덕열수추출물을 첨가했을 때 각각 4.2에서 13.8배, 3.1에서 6.9배의 증식효과를 나타냈다. 더덕열수추출물은 또한 RAW 264.7 세포주에 의한 $TNF-{\alpha}$ 나 IL-6 같은 사이토카인 생산을 자극하였다. RAW 264.7 세포를 lipopolysaccharide로 자극했을 경우, $TNF-{\alpha}$ 및 IL-6는 무첨가군에 비해 별다른 증진효과를 나타내지 않았으나, 자극하지 않은 RAW 264.7 세포에 대해서는 $TNF-{\alpha}$는 12.6에서 67.8배, Il-6는 2.8에서 10.1배 증가하였다. 이상의 결과로 더덕열수추출물을 이용하여 노화 및 질병에 따라 수반되는 면역력 감소를 방지하는 기능성식품 및 신소재로의 개발이 기대된다.

• 대한본초학회지
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• 제26권2호
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• pp.17-23
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• 2011

Objectives : This study aims at examining the effect of the fermentative extract of root of Sophorae Radix on the immuno-modulating activity. Methods : Cell viabilities were measured by MTT assay. Effect of SFS on nitric oxide(NO), hydrogen peroxide production from RAW 264.7 cells was accessed by Griess reagent assay. Effect of SFS on productions of inflammatory cytokines such as TNF-${\alpha}$, IL-6 in LPS-induced RAW 264.7 cells was accessed by a multiplex bead array assay based on xMAP technology. Results : The results of the experiment are as follows. 1. As a result of carrying out MTT assay to check the cellular toxicity of the fermentative extract of Sophorae Radix. There was not any excessive toxicity to the macrophage when the fermentative extract of root of Sophorae Radix was treated in different concentrations. 2. The fermentative extract of Sophorae Radix increased the generation of hydrogen peroxide in the macrophage and significantly restored the suppression of the generation of the hydrogen peroxide in the macrophage induced by LPS. 3. The fermentative extract of Sophorae Radix reduced the generation of NO in the macrophage and significantly suppressed the increase of the generation of NO in the macrophage induced by LPS. 4. The fermentative extract of Sophorae Radix significantly decreased the amount of TNF-${\alpha}$ generated in the macrophage induced by LPS when it was $25{\mu}g/mL$ or higher. Conclusion : These results suggest that SFS has anti-inflammatory moiety related with its inhibition of NO, hydrogen peroxide, TNF-${\alpha}$, IL-6, in macrophage led by LPS.

• Tuberculosis and Respiratory Diseases
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• 제47권2호
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• pp.172-183
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• 1999

연구배경: 산화질소(${\cdot}NO$)는 여러 세포에서 산화질소 합성효소(NOS)에 의해서 생산되며 다양한 병태생리과정에 관여한다. 여러 cytokine들이 iNOS의 발현을 촉진시키고 산화질소 생산을 증가시킴으로써 염증반응을 증폭시키고 세포와 조직손상을 초래한다고 알려진 바, 과산화수소($H_2O_2$)가 세포내 NOS의 발현과 산화질소형성에 미치는 영향을 알아보고자 하였다. 방법: 마우스 대식세포주 RAW264.7에 여러 가지 cytokine과 세균 내독소 (LPS)로 자극을 준 세포군 이에 더하여 $H_2O_2$, NOS 억제제 (L-NAME) 및 항산화제 (catalase)등을 사용하여 세포를 자극한 후 생성된 산화질소 산화물의 농도를 측정하고 Northern analysis로 iNOS mRNA의 발현정도를 보아 다음과 같은 성적을 얻었다. 결과: Cytokine과 LPS 자극군에서 대조군보다 ${\cdot}NO$ 생산이 높았고, 이 자극군에 $H_2O_2$를 추가로 자극하였을 때 ${\cdot}NO$생산이 2 배 이상 유의하게 높았다. Cytokine 자극군에서 $H_2O_2$의 자극 농도에 따른 ${\cdot}NO$생산은 $H_2O_2$의 농도가 증가할수록 유의하게 증가하였다. LPS와 IFN-$\gamma$ 자극군에서 L-NAME을 같이 자극시에 ${\cdot}NO$의 양은 L-NAME의 농도증가에 따라 유의하게 감소하였고, Cytokine 및 $H_2O_2$자극군에서도 추가로 자극한 L-NAME 의 농도증가에 따라 ${\cdot}NO$의 양은 유의하게 감소하였다. Cytokine과 $H_2O_2$ 자극균에 catalase를 같이 자극 하였을 때 ${\cdot}NO$의 양은 유의하게 감소했고, Mercaptoethanol과 phenanthroline을 전처치하고 LPS와 IFN-$\gamma$ 및 $H_2O_2$로 자극한 군에서 이들의 전처치한 농도가 높을수록 ${\cdot}NO$의 양은 유의하게 Cytokine자극군과 IFN-$\gamma$, LPS 자극군에 $H_2O_2$를 추가 자극 후 Northern analysis 결과 $H_2O_2$는 iNOS mRNA 발현을 현저히 증가시켰다. 결론: 이상의 결과로 과산화수소가 cytokine과 내독소 등으로 자극된 마우스 대식세포에서 산화질소생산에 유의한 증폭효과를 나타냈고, iNOS mRNA 의 발현도 증가시켰음을 확인할 수 있었다.

• Biotechnology and Bioprocess Engineering:BBE
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• 제6권1호
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• pp.72-74
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• 2001

Transgenic Nicotiana tabacum cells were cultivated for the production of murine granulocyte macrophage-colony stimulating factor (mGM-CSF) in both a stirred tank bioreactor and an airlift bioreactor with draft tube. Cell growth and mGM-CSF production in the airlift bioreactor were found to be better than those achieved in the stirred tank bioreactor. In the airlift bioreactor, 9.0g/L of cells and 2.2ng/mL of mGM-CSF were obtained (11.0g/L and 2.4ng/mL, respectively in shake flasks). Although the lag period was prolonged and mGM-CSF production was lowered by 33% in the stirred thank bioreactor as compared to the control culture, the maximum cell density was increased up to 12.0g/L due to better mixing by agitation at the higher cell density.

• Journal of Ginseng Research
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• 제39권4호
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• pp.365-370
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• 2015

Background: The beneficial effects of ginsenoside species have been well demonstrated in a number of studies. However, the function of ginsenoside Ro (GRo), an oleanane-type saponin, has not been sufficiently investigated. Thus, the aim of the present study was to investigate the anti-inflammatory effects of GRo in vitro using the Raw 264.7 mouse macrophage cell line treated with lipopolysaccharide (LPS), and to clarify the possible mechanism of GRo involving heme oxygenase-1 (HO-1), which itself plays a critical role in self-defense in the presence of inflammatory stress. Methods: Raw 264.7 cells were pretreated with GRo (up to $200{\mu}M$) for 1 h before treatment with 1 mg/mL LPS, and both cell viability and inflammatory markers involving HO-1 were evaluated. Results: GRo significantly increased cell viability in a dose dependent manner following treatment with LPS, and decreased levels of reactive oxygen species and nitric oxide. GRo decreased inflammatory cytokines such as nitric oxide synthase and cyclooxygenase-2 induced by LPS. Moreover, GRo increased the expression of HO-1 in a dose dependent manner. Cotreatment of GRo with tin protoporphyrin IX, a selective inhibitor of HO-1, not only inhibited upregulation of HO-1 induced by GRo, but also reversed the anti-inflammatory effect of GRo in LPS treated Raw 264.7 cells. Conclusion: GRo induces anti-inflammatory effects following treatment with LPS via upregulation of HO-1.

• Archives of Pharmacal Research
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• 제27권1호
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• pp.94-98
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• 2004

Effects of dopaminergic drugs on the degranulation of mast cells (RBL-2H3 cells) and the nitric oxide production from macrophage cells (RAW 264.7) were studied. Among the dopaminergic agonists and antagonists tested, bromocriptine, 7-OH-DPAT, haloperidol, and clozapine showed potent inhibitions of mast cell degranualtion ($IC_{50} value, 5 \mu$ M). However, these dopaminergic agents did not affect the tyrosine phosphorylations of the signaling components of the high affinity IgE receptor ($Fc\varepsilonRI$), such as Syk, $PLC\gamma1$, and $PLC\gamma2$.; This suggested that these signaling components were not involved in the inhibition of the mast cell degranulation by these compounds. On the other hand, dopamine, bromocriptine, 7-OH-DAPT, and haloperidol markedly inhibited the nitric oxide production from RAW 264.7 cells ($IC_{50}$ values, 10-20$\mu$M). Bromocriptine, a dopamine agonist that is routinely used for the treatment of Parkinsons disease, inhibited the expression of the inducible nitric oxide synthase at an early stage of the LPS-induced protein expression in a dose-dependent manner. The results suggested that these dopaminergic agents, when used for the treatment of dopamine receptors-related diseases, such as Schizophrenia or Parkinsons disease, might have additional beneficial effects.

• The Korean Journal of Physiology and Pharmacology
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• 제27권2호
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• pp.157-165
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• 2023

Triple-negative breast cancer (TNBC) is the most aggressive subtype of breast cancer and current therapeutic strategies are limited in their effectiveness. The expressions of Rab5 and the M2 tumor-associated macrophage marker CD163 in tissues were detected by Western blot. The migration and invasion of cells were determined using a Transwell assay. The expressions of the exosome markers were evaluated by Western blot. The polarization of human macrophages (THP-1) was determined by incubation of THP-1 cells with conditioned medium or exosomes collected from MDA-MB-231 cells with indicated transfections or by a coculture system of THP-1 and MDA-MB-231 cells. The M1 and M2 macrophage markers were evaluated by qRT-PCR. The expression of Rab5 in TNBC was significantly higher than that in normal breast tissue. Rab5 expressions in triple-negative and luminal A breast cancer were higher than those in other molecular subtypes. Higher CD163 expression was observed in triple-negative breast cancer and in triple-negative and luminal B subtypes. Rab5 knockdown suppressed but Rab5 overexpression promoted the migration and invasion capacity of MDA-MB-231 cells. The levels of CD63 and CD9 in the medium of Rab5 knockdown cells were lower than those in control cells, whereas higher levels of CD63 and CD9 were observed in Rab5 overexpression cells. Rab5 knockdown decreased the excretion but did not alter the diameter of the exosomes. Knockdown of Rab5 facilitated the anti-tumor polarization of macrophages, which was partially reversed by Rab5 overexpression. Therefore, Rab5 is expected to be a potential therapeutic target for triple-negative breast cancer.