• 한국식품위생안전성학회지
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• 제27권4호
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• pp.406-414
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• 2012

본 연구에서는 fucoidan이 macrophage의 활성과 항암효과를 확인하기 위하여 수행되었다. Fucoidan을 Raw 264.7 세포에 처리한 후 MTT assay로 측정한 결과 고농도 $100{\mu}g/mL$까지 세포독성은 없었으며, NO 및 TNF-${\alpha}$의 분비를 농도 유의적으로 증가시켰다. 또한 AGS 위암 세포 성장저해효과를 확인하기 위하여 MTT assay를 하였고 그 결과 농도와 시간 의존적으로 암 세포 성장이 유의적으로 감소하였다. Apoptosis로 인해 암 세포 성장이 감소하였는지 확인하기 위하여 DAPI 염색을 한 결과 apoptotic body와 세포질 응축이 시간 의존적으로 증가하는 것을 확인하였다. 또한 fucoidan은 미토콘드리아의 투과율을 향상시키며 미토콘드리아에서 방출되는 cytochrome c의 발현을 증가시켰다. Western blotting의 결과 시간 의존적으로 anti-apoptotic 분자인 Bcl-2와 XIAP 발현 감소와 반대로 pro-apoptotic 분자인 Bax 발현이 증가하였다. Cleaved-caspase-9의 발현이 증가하였으며 Akt의 인산화는 시간 의존적으로 감소하였다. Caspase 억제제인 z-VAD-FMK 처리 시 Bax, caspase-9의 발현을 감소시켜 apoptosis 유도를 억제하였으며 이러한 결과는 caspase가 apoptosis 유도에 중요한 역할을 하는 것으로 나타낸다. 본 실험의 AGS 위암 세포주에서 대조군에 비하여 fucoidan 처리군에서 면역세포 활성 및 AGS 위암 세포주에서 caspase 활성을 통해 apoptosis를 유도하는 것으로 사료된다.

• 대한본초학회지
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• 제34권1호
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• pp.23-31
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• 2019

Objectives : The aim of this study was to investigate the effect for allergic-inflammation of Fritillariae Thunbergii Bulbus (FTB) on HaCaT cells and RBL2H3 cells. Methods : To investigate the effects of FTB for anti-inflammation in HaCaT cells, the cells were pretreated with FTB for 1h and then stimulated with $TNF-{\alpha}/IFN-{\beta}$ for 24h. Then thymus and activation-regulated chemokine (TARC) and Macrophage-derived chemokine (MDC) levels were analyzed with ELISA kit. Also to investigate the effect of skin barrier protein, the cells were treated with FTB of various concentrations, and then cells were harvested, expressions of skin barrier protein were measured with RT-PCR. To investigate the effects of FTB for anti-allergy in RBL2H3 cells, the cells were pre-treated with FTB for 1h, and then stimulated with A23187 for 30 min. ${\beta}$-hexosaminidase, IL-4 and $TNF-{\alpha}$ were measured using cultured media. The cells were harvested to analyze the mechanism of the effect for FTB via Western blot. Results : FTB did not show cytotoxicity in HaCaT and RBL2H3. In HaCaT cells, FTB significantly suppressed the expression of TARC, MDC at a dose-dependent manner and markedly increased formation of the skin barrier proteins. In RBL2H3 cells, FTB decreased release of the ${\beta}$-hexosaminidase, IL-4 and $TNF-{\alpha}$ in RBL2H3 through inhibition of the phosphorylation of JNK and p38, which are include in the signaling mechanism of MAPK Conclusion : These results indicate that FTB has an anti-inflammatory effect on the allergic response through blocking MAPK pathway. This suggest that FTB could be a therapeutic agent for allergic response.

• 생명과학회지
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• 제21권5호
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• pp.678-683
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• 2011

본 연구에서는 울릉도 특산 약용 작물 추출물이 항염증에 대한 실험연구가 이루어져 있지 않은 것에 착안하여 LPS에 의해 활성화된 대식세포로부터 유도되는 염증반응에 대한 억제효과를 조사하였다. 울릉도 자생 식물인 곰취, 미역취, 삼나물의 세가지 식물 추출물을 이용하여 피부 염증에 대하여 연구를 하였다. 산화질소와 cytokine의 생산은 면역세포의 대표적인 염증인자이다. 세포는 LPS 처리 후 한 시간 뒤에 곰취, 미역취, 삼나물 70% 아세톤 추출물을 처리를 하였다. 세포 독성이 나타나지 않는 농도인 1, 10, 100 ug/ml를 사용하였다. 곰취, 미역취, 삼나물 70% 아세톤 추출물은 NO, $PGE_2$, TNF-${\alpha}$, IL-1${\beta}$, IL-6, iNOS, COX-2의 생성을 저해 시켰다. $PGE_2$는 100 ug/ml의 농도에서 60%에 가까운 저해율을 나타내었다. iNOS와 COX-2 역시 100 ug/ml의 농도에서 각각 54%, 65%가 저해가 되었다. 게다가 곰취, 미역취, 삼나물 70% 아세톤 추출물은 염증성 사이토 카인인 TNF-${\alpha}$, IL-1${\beta}$, IL-6의 생성을 감소 시켰다. 이러한 결과로 곰취, 미역취, 삼나물 추출물은 염증 예방과 치료에 효과적임을 확인할 수 있었다.

• Archives of Pharmacal Research
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• 제25권6호
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• pp.895-902
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• 2002

Monocytes and macrophages playa major role in defense mechanism of the host response to tumor, in part through the secretion of several potent products and macrophage cytokines. Monocytes and tissue macro phages produce at least two groups of protein mediators of inflammation, interleukin 1 (IL-1) and tumor necrosis factor (TNF). Recent studies emphasizes that TNF and IL-1 modulate the inflammatory function of endothelial cells, leukocytes, and fibroblasts. In this study, our work is directed toward studying the in vitro effects of Korean propolis on the ability to induce cellular and secretory responses in murine macrophage cell line, RAW 264.7. It was found that Water Extract of Korean Propolis (WEP) could activate macro phages by producing cytokines. The production of the macrophage cytokines, IL-1 and TNF-$\alpha$, by RAW 264.7 treated with WEP was examined from 2.5 $\mu\textrm{g}$/ml up to 25 $\mu\textrm{g}$/ml with dose dependent manner. Nitric oxide (NO) production was also increased when cells were exposed to combination of LPS and WEP from 2.5 $\mu\textrm{g}$/ml up to 25 $\mu\textrm{g}$/ml. At high dose of WEP (50 to 100 $\mu\textrm{g}$/ml) used to prescribe for anti-inflammatory and analgesic medicine showed inhibition of NO production in LPS-stimulated macrophage. Besides cytokine production, NO release, surface molecule expression and cell morphologic antigen expression were increased in response to the stimulation by WEP. These results suggested WEP may function through macrophage activation.

• 한국축산식품학회지
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• 제30권5호
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• pp.739-745
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• 2010

The objective of the current study was to compare the abilities of undigested and enzymatically digested bifidobacteria to induce nitric oxide and cytokine release in RAW 264.7 macrophage cells. Nine different Bifidobacterium strains derived from herbivorous animals were digested with pepsin and then pancreatin, and the precipitates and supernatants were acquired via centrifugation. The RAW 264.7 cells were incubated with whole cells, the precipitate, or the supernatant, and the macrophage culture supernatants were analyzed with respect to the induction of nitric oxide and cytokines. Pronounced increases in the production of nitric oxide, interleukin (IL)-$1{\beta}$, IL-6, IL-12, and tumor necrosis factor-$\alpha$ (TNF-$\alpha$) were observed when cultured with whole cells and the precipitates. It is noteworthy that the precipitates in most of the Bifidobacterium strains evidenced a trend toward superior IL-12 release compared with whole cells. The results showed that both whole cells and digested Bifidobacterium sp. are effective at stimulating RAW 264.7 cells to induce the production of nitric oxide and cytokines. The pepsin-pancreatin system used in the current study may be useful in unraveling the mechanism by which ingested lactic acid bacteria modulate the induction of macrophage mediators at the cellular level.

• 대한한의학회지
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• 제20권1호
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• pp.11-21
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• 1999

Dansam, the root of Salvia miltiorrhiza BGE. (Labiatae), has a bitter and a slightly 'cold' property. and is nontoxic. It has been used for the treatment of diseases such as tumors, bruises, gynecologic diseases, menoxenia, anemia and so forth. As an oriental medicine pathway, anal therapy has many kinds of treatments, Retention enema is one the most useful, advantageous methods of anal therapy. This experiment was performed in order to study the effect of an aqueous extract of Salviae radix root(SRRAE) on Immediate type allergic reaction by Retention enema. The results were obtained as follows, 1. SRRAE inhibited compound 48/80-induced Immediate type allergic reaction 100% with the dose of 0.1g/kg by anal treatment. However, SRRAE showed no significant inhibitory effect on the same reaction by oral treatment. When mice were pretreated with SRRAE at a concentration ranging from 0.001 to 1.0g/kg, by intra-anal treatment, the serum histamine levels were reduced in a dose-dependent manner. 2. SRRAE also inhibited by 79.8% local cutaneous allergic reaction activated by anti-dinitrophenyl (DNP) IgE. 3. SRRAE close-dependently inhibited the histamine release from rat peritoneal mast cells(RPMC) by anti-DNP IgE., but SRRAE not inhibited compound 48/80-induced histamine release. 4. SRRAE dose-dependently inhibited tumor necrosis $factor-\;{\alpha}$ $(TNF-\;{\alpha}$) production from RPMC by anti-DNP IgE. Moreover, the level of cAMP in RPMC, when SRRAE was added, significantly increased compared with that of a normal control. According to the above results, anal therapy(Retention enema) of SRRAE may be beneficial in the treatment of systemic and local Immediate type allergic reactions.

• 동의생리병리학회지
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• 제23권1호
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• pp.206-213
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• 2009

본 연구는 RBL-2H3 세포에서 고삼의 NF-${\kappa}B$ (p65) 활성 억제를 통한 과립감소와 전염증성 시토카인 억제 효과에 관한 것으로 주요 내용은 다음과 같다. 본 연구에서는 PMA와 A23187로 유발된 흰쥐 백혈병(RBL-2H3) 세포에서 고삼의 항알레르기 효과에 대하여 알아보았다. 고삼은 투여량에 따라 $\beta$-hexosaminidase의 방출과 TNF-$\alpha$, IL-4, COX-2 등의 생성과 발현을 억제하였다. 실험결과 고삼은 $NF-{\kappa}B$ (p65)의 조절을 통하여 항알레르기 효과를 나타내었는데 이는 $I{\kappa}B-{\alpha}$ 저해의 억제와 항염증 시토카인 발현 억제와도 관계가 있다는 내용이다.

• 대한한방내과학회지
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• 제29권2호
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• pp.318-333
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• 2008

Objectives : In this study, the author tried to examine whether Cheogjogupye-tang (淸燥救肺湯, CGPT) and Yieum-jeon (理陰煎, YEJ) significantly affect in vitro and in vivo mucin secretion, MUC5AC gene expression in airway epithelial cells and contractility of isolated tracheal smooth muscle of rabbit. Materials and Methods : For in vitro experiment, confluent hamster tracheal surface epithelial (HTSE) cells were chased for 30 minutes in the presence of CGPT and YEJ to assess the effects of the agents on mucin secretion by enzyme-linked immunosorbent assay (ELISA), with removal of oriental herbal medicine extract from each agent-treated sample by centrifuge microfilter. Also, the effects of the agents on TNF-alpha or EGF-induced MUC5AC gene expression in human airway epithelial cells (NCI-H292) were investigated. Possible cytotoxicities of the agent were assessed by examining both LDH release from HTSE cells and the rate of survival and proliferation of NCI-H292 cells. For in vivo experiment, hypersecretion of airway mucin and goblet cell hyperplasia was induced by exposure of rats to $SO_2$ over 3 weeks. Effects of CGPT and YEJ orally administered for 1 week on in vivo mucin secretion from tracheal goblet cells of rats and hyperplasia of goblet cells were assessed using ELISA and histological analysis after staining the epithelial tissue with alcian blue, respectively. Also, the effects of CGPT and YEJ on contractility of isolated tracheal smooth muscle were investigated. Results : (1) CGPT significantly inhibited in vitro mucin secretion from cultured HTSE cells. However, YEJ did not affect in vitro mucin secretion; (2) CGPT and YEJ did not affect hypersecretion of in vivo mucin and hyperplasia of tracheal goblet cells; (3) CGPT and YEJ slightly increased the expression levels of TNF-alpha or EGF-induced MUC5AC gene in NCI-H292 cells; (4) CGPT and YEJ inhibited acetylcholine-induced contraction of isolated tracheal smooth muscle of rabbit; (5) CGPT and YEJ did not affect LDH release from HTSE cells and the survival and proliferation of NCI-H292 cells. Conclusion : The results from the present study suggest that CGPT and YEJ mainly affect the expression of mucin gene rather than secretion of mucin and do not show remarkable cytotoxicity to respiratory epithelial cells.

• 한국식품영양과학회지
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• 제43권11호
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• pp.1635-1641
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• 2014

본 연구는 고욤잎 유래 EA 분획물의 항산화 및 항가려움 활성을 조사하였다. EA 분획물은 DPPH 라디칼 소거 활성이 수용성 및 MeOH 추출물보다 매우 우수하였을 뿐만 아니라 합성 항산화제인 BHT보다 현저히 높았다. 또한 ABTS 라디칼 소거 활성도 수용성과 MeOH 추출물뿐만 아니라 Trolox보다 현저히 높았다. 본 연구에 사용된 EA 분획물과 수용성 및 MeOH 추출물은 세포에 대한 독성이 없었다. 또한 EA 분획물은 농도에 의존적으로 활성화된 비만세포로부터 히스타민 방출 억제와 TNF-${\alpha}$ 생성 억제 효과가 있었으며, 같은 농도에서 수용성 및 MeOH 추출물보다 억제 효과가 우수하였다. 더욱이 EA 분획물은 농도 의존적으로 compound 48/80으로 유도된 가려움증을 효과적으로 억제하는 활성이 있었고, 같은 농도에서 항가려움 약제로 알려진 azelastine보다는 그 효과가 낮았지만 수용성 및 MeOH 추출물뿐만 아니라 methysergide보다 우수한 항가려움 효과를 발휘하였다. 앞으로 이러한 항가려움 효과에 대한 EA분획물의 분자적 작용기전을 규명해야 할 필요가 있으며, 고욤잎 유래 EA 분획물은 강력한 항산화 및 항가려움 활성을 지닌 우수한 소재라 사료된다.

• 한국키틴키토산학회지
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• 제23권4호
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• pp.277-284
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• 2018

본 연구에서는 제주 자생해조류인 감태로부터 제조한 ethyl acetate 분획물의 비만 세포에 대한 항알러지 활성에 관한 연구를 수행하였다. IgE/BSA로 활성화시킨 비만 세포에서 EC-EtoAc는 세포 독성 없이 비만 세포의 탈과립 지표로 사용되는 ${\beta}$-hexosaminidase의 분비 뿐만 아니라 알러지 유발성 케모카인(TARC)과 사이토카인(IL-$1{\beta}$, IL-4, IL-6, IL-10, IL-13, IFN-${\gamma}$ 및 TNF-${\alpha}$)의 mRNA 발현을 억제하였다. 또한, EC-EtoAc는 비만 세포의 표면 리셉터인 $Fc{\varepsilon}RI$의 발현 및 IgE와 $Fc{\varepsilon}RI$의 결합능을 감소시켰다. 이 모든 결과로부터, 감태 유래 분획물인 EC-EtoAc가 활성화된 비만 세포에서 $Fc{\varepsilon}RI$의 발현 및 IgE와 $Fc{\varepsilon}RI$의 결합능을 감소시킴으로써 알러지 유발성 매개 인자들의 분비와 발현 등을 감소시켜 항알러지 효능을 가진다는 것을 알 수 있었다. 또한, 이 연구 결과는 EC-EtoAc가 항알러지 효능을 보이는 천연 소재로서 이용가치가 있다는 것을 제시한다.