• Mycobiology
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• 제38권2호
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• pp.144-148
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• 2010

$\beta$-Glucans have been known to exhibit antitumor activities by potentiating host immunity by an unknown mechanism. The C-type lectin dectin-1, a $\beta$-glucan receptor, is found on the macrophage and can recognize various $\beta$-glucans. Previously, we demonstrated the presence of $\beta$-glucan receptor, dectin-1, on the Raw 264.7 cells as well as on murine mucosal organs, such as the thymus, the lung, and the spleen. In order to investigate immunopotentiation of innate immunity by $\beta$-glucan, we stimulated a murine macrophage Raw 264.7 cell line with $\beta$-glucans from Pleurotus ostreatus, Saccharomyces cerevisiae, and Laminaria digitata. Then, we analyzed cytokines such as tumor necrosis factor (TNF)-$\alpha$ and interleukin (IL)-6 by reverse transcription-polymerase chain reaction (RT-PCR). In addition we analyzed gene expression patterns in $\beta$-glucan-treated Raw 264.7 cells by applying total mRNA to cDNA microarray to investigate the expression of 7,000 known genes. When stimulated with $\beta$-glucans, the macrophage cells increased TNF-$\alpha$ expression. When co-stimulation of the cells with $\beta$-glucan and lipopolysaccharide (LPS), a synergy effect was observed by increased TNF-$\alpha$ expression. In IL-6 expression, any of the $\beta$-glucans tested could not induce IL-6 expression by itself. However, when co-stimulation occurred with $\beta$-glucan and LPS, the cells showed strong synergistic effects by increased IL-6 expression. Chip analysis showed that $\beta$-glucan of P. ostreatus increased gene expressions of immunomodulating gene families such as kinases, lectin associated genes and TNF-related genes in the macrophage cell line. Induction of TNF receptor expression by FACS analysis was synergized only when co-stimulated with $\beta$-glucan and LPS, not with $\beta$-glucan alone. From these data, $\beta$-glucan increased expressions of immunomodulating genes and showed synergistic effect with LPS.

• 대한본초학회지
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• 제24권1호
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• pp.169-178
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• 2009

Objectives : In this study, the effects of ethylacetate extract of Ostericum koreanum on inflammation in RAW264.7 cells were investigated. Methods : Dried roots of Ostericum koreanum was extracted with 80% methanol for 24 h, and then fractionated with n-butanol, n-hexan and ethylacetate. RAW264.7 cells, a mouse macrophage line were incubated with different concentrations of the extract for 30 min and then stimulated with LPS at indicated times. Cell toxicity was determined by MTT assay. The concentrations of nitric oxide (NO) and prostaglandin E2 ($PGE_2$) were measured by Griess assay and enzyme immunoassay (EIA), respectively. The expressions of inducible nitric oxide synthease (iNOS) and cyclooxyganase (COX) -2 mRNA and protein were determined by RT-PCR and Western blot. Results : The methanol extract of Ostericum koreanuman and its fractions were significantly inhibited the NO and PGE2 productions in LPS-stimulated RAW264.7 cells. Among the fractions of Ostericum koreanuman the ethylacetate fraction was more strongly inhibited NO and $PGE_2$ productions compared with other fractions. The ethylacetate fraction was also suppressed LPS-induced mRNA expressions of iNOS and its protein levels in RAW264.7 cells. Conclusions : This study suggests that the ethylacetate fraction of Ostericum koreanum may have an anti-inflammatory property through suppressing inflammatory mediator productions in activated macrophages, suggesting have a therapeutic potential for the treatment of various inflammatory diseases.

• Mycobiology
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• 제40권1호
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• pp.47-52
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• 2012

$Elfvingia$ $applanata$, a medicinal mushroom belonging to Basidiomycota, has been used in the effort to cure cancers of the esophagus and stomach, and is also known to have inhibitory effects on hepatitis B virus infection. The hot water soluble fraction (as Fr. HW) was extracted from fruiting bodies of the mushroom. $In$ $vitro$ cytotoxicity tests showed that hot water extract was not cytotoxic against cancer cell lines such as Sarcoma 180, HT-29, HepG2, and TR at concentrations of 10-2,000 ${\mu}g/mL$. Intraperitoneal injection with Fr. HW resulted in a life prolongation effect of 45.2% in mice previously inoculated with Sarcoma 180. Treatment of Fr. HW resulted in a 2.53-fold increase in the numbers of murine spleen cells at a concentration of 50 ${\mu}g/mL$, compared with control. Incubation of murine spleen cells with Fr. HW at a concentration of 500 ${\mu}g/mL$ resulted in improved immune-potwntiating activity of B lymphocytes through an 8.3-folds increase in alkaline phosphatase activity, compared with control. Fr. HW generated 12.5 ${\mu}M$ of nitric oxide (NO) when cultured with RAW 264.7, a mouse macrophage cell line, at the concentration of 50 ${\mu}g/mL$, while lipopolysaccharide, a positive control, produced 15.2 ${\mu}M$ of NO. Therefore, the results suggested that antitumor activities of Fr. HW from $E.$ $applanata$ might, in part, be due to host mediated immunostimulating activity.

• Journal of Nutrition and Health
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• 제48권6호
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• pp.459-467
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• 2015

본 연구는 maltodextrin cyclodextrin에 의해 형성된 고분자 미셀에 나노캡슐화 시킨 리코펜과 일반 리코펜을 LPS로 염증을 유도시킨 RAW 264.7 대식세포주에 다양한 농도 (0~20)로 처리하여 염증매개 사이토카인 유전자 발현과 단백질 생성, 염증매개 효소인 iNOS, COX-2 mRNA 유전자 발현에 미치는 영향을 비교하였다. 나노캡슐화된 리코펜은 염증세포의 증식억제, 염증매개 사이토카인, 세포침식등의 염증으로 진행되는 데 관여하는 IL-6, IL-$1{\beta}$, TNF-${\alpha}$ 등의 사이토카인과 iNOS, COX-2 유전자 발현을 일반리코펜에 비해 효과적으로 억제하고 사이토카인 단백질 생성억제를 통해 염증억제 효과를 증진시키는 것이 관찰되었다. 따라서 지용성으로 생체이용성이 낮은 생리활성물질의 건강증진효과를 개선시키기 위해 나노캡슐화는 좋은 모델이 될 수 있을 것으로 사료된다.

• 동의생리병리학회지
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• 제28권3호
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• pp.330-336
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• 2014

This study aimed at examining the anti-inflammatory effects of Scutellariae Radix & Lonicerae Caulis water extract(SC). RAW 264.7 mouse macrophage cells were treated with $25{\sim}200{\mu}g/m{\ell}$ SC for 24 hours. Cell viability was then measured using MTT assays. The nitric oxide(NO) production and the creation of several cytokines in LPS-stimulated RAW 264.7 cells were investigated. SC inhibited significantly increasing the production of NO in LPS-induced RAW 264.7 cell at the density of 25, 50 and $200{\mu}g/m{\ell}$. SC inhibited significantly the TNF-${\alpha}$ of the RAW 264.7 cell induced by LPS at the density of $50{\mu}g/m{\ell}$. SC inhibited significantly the MIP-$1{\alpha}$ of the RAW 264.7 cell induced by LPS at the density of 25, 50 and $100{\mu}g/m{\ell}$. SC inhibited significantly the MIP-$1{\beta}$, MIP-2 at the density of 50, $100{\mu}g/m{\ell}$ in the RAW 264.7 cell increased by LPS, respectively. SC did not affect the production levels of VEGF in RAW 264.7 cell. As a result, SC significantly inhibited the inductions of MIP-$1{\alpha}$, MIP-$1{\beta}$, MIP-2 and NO in LPS-induced RAW 264.7 cell without causing the toxicity. These results signify that SC has anti-inflammatory effects on controlling the over inflammatory reaction on the RAW 264.7 cell.

• 한방안이비인후피부과학회지
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• 제20권2호통권33호
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• pp.10-35
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• 2007

Objectives : To investigate the effects of Ohbaesangami (OBSGM) on mucosa and skin diseases, anti-microbial and anti-inflammatory tests were performed using several in vitro test models. Results : In anti-microbial test, OBSGM showed the slight inhibitory effect against Propionibacterium acnes (P. acnes) and Staphylococcus aureus (S. aureus). In anti-oxidant test, OBSGM showed the potent radical scavenging activity. In anti-inflammatory test, OBSGM weakly inhibited the lipopolysaccharide (LPS)-induced nitric oxide(NO) release from the RAW 264.7 macrophage cells. OBSGM also inhibited the LPS-induced $interleukin-1{\beta}(IL-1{\beta})$ and cyclooxygenase-2 (COX-2) expressions. The inhibitory effects of OBSGM on macrophage activation was via the inhibition of $NF-{\kappa}B$, evidenced by transient transfection assay. Furthermore, OBSGM markedly inhibited the activation of Jun-N-terminal kinase (JNK) and p38 MAP kinase in RAW 264.7 cells. In skin wrinkle formation assay, OBSGM strongly inhibited collagnease and elastase, whose activities are tightly related with the wrinkle formation. In addition, OBSGM inhibited the activities of MMP-1, MMP-2 on the mRNA levels in RAW 264.7 cells. However, OBSGM did not show an inhibitory potential on tyrosinase activity and melanin synthesis, indicating that it could not be applicable for skin whitening. Conclusion : These results suggest that the anti-inflammatory effect of OBSGM may be due to its inhibitory potentials on the macrophage activation. And, the anti-wrinkle effects of OBSGM may be due to its inhibitory potential on the collagnease and elastase activities. Therefore, OBSGM could be applicable for the treatment of mucosa and skin diseases.

• 한국식품과학회지
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• 제44권2호
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• pp.228-234
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• 2012

현재 우리나라에서 시판되고 있는 대부분의 양조간장은 일본의 $koji$ 방식으로 제조되고 있는 반면, 재래식 조선간장은 자연적으로 미생물이 접종 및 발효된 메주를 이용하여 제조되고 있다. 본 연구는 일본식으로 만들어진 양조간장과 우리나라 전통방식으로 만들어진 재래식 조선간장으로부터 얻어진 다당의 특성과 면역증진활성을 비교하였다. 조선간장 유래 다당인 KTSP-0는 2-keto-3-deoxy-D-$manno$-2-octulosonic acid(KDO)(1.1%)의 구조를 가지는 rhamnogalacturonan II(RG-II)가 있음을 확인하였다. 양조간장 유래 다당 CSP-0와 KTSP-0의 보체계 활성화능을 측정한 결과 두 시료 모두 농도의존적으로 증가하는 보체계 활성을 가지고 있었으나, $1,000{\mu}g/mL$ 농도에서 KTSP-0는 64.7%, CSP-0는 56%의 활성을 나타내었다. C3의 보체계 활성 경로를 확인하기 위해 2차원 면역전기영동을 행하였다. 그 결과, CSP-0는 고전경로로만 보체계를 활성화 시키는 반면, KTSP-0는 고전경로와 부경로 모두를 통해 보체계를 활성화 시키는 것으로 나타났다. CSP-0와 KTSP-0는 비장 유래세포와 macrophage에 대한 세포독성이 없음이 확인되었으며, KTSP-0는 활성화된 복강유래 macrophage에서 IL-6는 $8-1,000{\mu}g/mL$의 비교적 넓은 범위에서, IL-12는 $40{\mu}g/mL$의 농도에서 cytokine의 분비를 증가시켰다. 본 연구를 통해 양조간장보다 조선간장이 보체계 활성화를 통해 높은 면역 증강 효과를 가지고 있는 것으로 확인되었다.

• 동의생리병리학회지
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• 제18권6호
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• pp.1622-1627
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• 2004

Nephrite has been widely used as a medicinal mineral resource to treat a numerous chronic diseases and to replenish vital essence and blood in the Korean traditional medicine. However, as of yet, there is little understanding of the pharmacological and biochemical mechanisms of its therapeutic effects as regards anti-inflammation. We therefore examined whether nephrite represses the expression of major inflammation mediators, such as interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), inducible nitric oxide synthase (iNOS), and cyclooxygenase-2 (COX-2), in LPS-stimulated murine macrophage cell line, RAW264.7 by using RT-PCR. The powder suspension and water extracts of nephrite significantly inhibited the mRNA expression of the mediators, despite a little toxic effects on growth of RAW 264.7 cells within the concentration range tested. These experimental results suggested that the nephrite can be utilized as a functional mineral exerting the anti-inflammation medicinal effect.

• 한국식품과학회지
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• 제34권3호
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• pp.510-517
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• 2002

밀 배아에서 제조된 arabinoxylan(A1: low MW, A2: medium MW, A3, high MW)의 면역세포 활성화 작용을 invitro에서 마우스 비장 림프구와 복강 대식세포를 대상으로 관찰하여 다음과 같은 결과를 얻었다. 밀 arabinoxylan으로 처리된 마우스 비장 림프구의 생존능을 살펴보았을 때 50, $100\;{\mu}g/mL$ 농도에서 A3이 비장 림프구의 생존을 증가시켰고 비장 림프구에 $10\;{\mu}g/mL$ LPS를 첨가하여 활성화된 상태에서 $20\;{\mu}g/mL$ arabinoxylan을 처리한 결과 A1과 A3이 대조군에 비해 비장 림프구의 생존을 유의적으로 증가시켰다(p<0.05). 마우스 복강 대식세포의 생존율 관찰한 결과 $10{\sim}100\;{\mu}g/mL$ 농도에서 A1과 A3은 대식세포의 생존을 유의적으로 증가시켰다(p<0.05). 대식세포의 암세포 살해능을 살펴보았을 때 $5\;{\mu}g/mL$ 농도의 A3이 암세포독성을 유의적으로 증가시켰으며, phagocytic index를 측정한 결과 arabinoxylan을 $20\;{\mu}g/mL$ 농도로 처리했을 때, 대조군에 비하여 유의적인 증가 효과를 나타내어 밀 arabinoxylan이 대식세포의 식세포 작용을 증가시킴을 알 수 있었다(p<0.05). 또한 arabinoxylan은 대식세포의 lysosomal phosphatase와 myeloperoxidase 활성을 유의적으로 증가시켰으며(p<0.05) NO 생성을 감소시키는 경향을 나타내었다. 대식세포에서 분비되는 $H_2O_2$의 양을 측정한 결과, arabinoxylan은 유의적인 $H_2O_2$ 생성 증가를 나타내었고(p<0.05), NBT 환원법으로 대식세포의 $O_2$ 생성 지표를 측정하였을 때, arabinoxylan은 유의적으로 대식세포의 NBT 환원을 증가시켰다(p<0.05). 이상의 결과로 미루어 볼 때, 밀 arabinoxylan의 면역세포 활성화 효과는 대식세포에서 분비되는 lysosomal enzyme 및 반응성산소종(ROI)의 생성과 밀접한 관련이 있는 것으로 생각된다.

• Journal of Acupuncture Research
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• 제21권3호
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• pp.107-119
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• 2004

Objective : The purpose of this study was to investigate the effect of Bee Venom and melittin on the lipopolysaccharide(LPS) and sodium nitroprusside(SNP)-induced expressions of Cell viability, nitric oxide(NO), inducible nitric oxide synthase(iNOS), extra-signal response kinase(ERK), jun N-terminal Kinase(JNK) and p38 kinase(p38)- mitogen activated protein kinase(MAPK) Family- in RAW 264.7 cells, a murine macrophage cell line. Methods : The expressions of cell viability by MTT assay, NO by Nitrite assay and iNOS, ERK, JNK and p38 were determined by Western blotting. Results : 1. Compared with the control group, 0.5, 1, $5{\mu}g/m{\ell}$ bee venom and 5, $10{\mu}g/m{\ell}$ melittin increased cell viability of RAW 264.7 induced by LPS and SNP significantly respectively. 2. Compared with the control group, 0.5, 1, $5{\mu}g/m{\ell}$ bee venom and 5, $10{\mu}g/m{\ell}$ melittin inhibited expression of NO induced by LPS and SNP significantly respectively. 3. Compared with the control group, 1, $5{\mu}g/m{\ell}$ bee venom and 5, $10{\mu}g/m{\ell}$ melittin inhibited expression of iNOS induced by LPS significantly and 0.5, 1, $5{\mu}g/m{\ell}$ bee venom and 5, $10{\mu}g/m{\ell}$ melittin inhibited expression of iNOS induced by SNP significantly. 4. Compared with the control group, the expression of ERK induced by LPS and SNP decreased significantly in the treatment groups of $5{\mu}g/m{\ell}$ bee venom and 5, $10{\mu}g/m{\ell}$ melittin, which of p-ERK by LPS also did in 1, $5{\mu}g/m{\ell}$ bee venom and 5, $10{\mu}g/m{\ell}$ melittin, but which of p-ERK by SNP did not decrease. 5. Compared with the control group, the. expression of JNK induced by LPS and SNP decreased significantly in the treatment groups of 5, $10{\mu}g/m{\ell}$ melittin, which of p-JNK by LPS in 5, $10{\mu}g/m{\ell}$ melittin and by SNP in $1{\mu}g/m{\ell}$ bee venom and $10{\mu}g/m{\ell}$ melittin decreased significantly. 6. Compared with the control group, the expression of p38 induced by LPS did not have significant difference, which induced by SNP decreased significantly in the treatment groups of 1, $5{\mu}g/m{\ell}$ bee venom and 5, $10{\mu}g/m{\ell}$ melittin. p-p38 induced by LPS decreased significantly in the treatment group of $10{\mu}g/m{\ell}$ of melittin, which induced by SNP also decreased significantly in 0.5, 1, $5{\mu}g/m{\ell}$ bee venom and 5, $10{\mu}g/m{\ell}$ melittin.