• 대한한방부인과학회지
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• 제20권1호
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• pp.144-160
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• 2007

목 적 : 이 연구는 천식, 기관지염, 폐렴, 결핵, 산후감모 등의 호흡기 질환에 사용되는 청폐화담탕(淸肺化痰湯)의 항염작용(抗炎作用)의 효과에 대해 알아보기 위해 시행되었다. 방 법 : 청폐화담탕(淸肺化痰湯)의 항염작용(抗炎作用)의 효과를 평가하기 위해 세포독성에 미치는 영향, NO, TNF-${\alpha}$, IL-l${\beta}$, IL-6 생성량에 미치는 영항, TNF-${\alpha}$, IL-l${\beta}$, IL-6 유전자 발현에 미치는 영향, iNOS, 염증cytokine 유전자 및 단백질 발현에 미치는 영향, PGE$_2$ 합성에 미치는 영향 및 NF-${\kappa}$B 활성에 미치는 영향에 대한 실험평가를 하였다. 결 과 : 청폐화담탕(淸肺化痰湯)은 MTT 분석을 통한 RAW 264.7 세포주의 생존력 평가에서 세포독성이 없었고, LPS로 유도된 RAW 264.7 세포주에서 NO 생성량을 농도 의존적으로 억제하였다. 청폐화담탕(淸肺化痰湯)은 400 g/ml 농도에서 LPS로 유도된 RAW 264.7 세포주에 대해 TNF-${\alpha}$, IL-1${\beta}$, IL-6 생성량을 각각 41.86${\pm}$2.26 %, 61.11${\pm}$2.54 %, 55.33${\pm}$3.65 % 억제하였으며 TNF-${\alpha}$, IL-1${\beta}$ 및 IL-6 유전자 발현을 농도 의존적으로 억제하였고, LPS로 유도된 RAW 264.7 세포주에서 iNOS와 COX-2 유전자 및 단백질 발현은 농도 의존적으로 억제하였다. 또한 그 농도에 따라 PGE$_2$ 생성량이 현저하게 억제하였고, LPS로 유도된 NF-${\kappa}$B 전사활성을 농도 의존적으로 억제함으로써 iNOS와 염증Cytokine 유전자 발현을 하향조절 하였다. 결 론 : 이상의 실험을 통해 청폐화담탕(淸肺化痰湯)은 LPS로 유도된 macrophage에서 NO와 염증Cytokine 생성량을 억제하였고 murine macrophage에서 NF-${\kappa}$B 활성을 억제함으로써 iNOS와 염증Cytokine 유전자 발현을 하향조절 하였다. 이러한 청폐화담탕(淸肺化痰湯)의 항염작용으로 천식, 기관지염, 폐렴, 결핵, 산후감모 등의 호흡기 질환에 응용할 수 있으리라 사료된다.

• 한국식품영양학회지
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• 제29권3호
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• pp.431-437
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• 2016

본 연구는 전통약재로 주로 사용되는 구절초에서 분리한 다당류(CZPS)가 선천 및 적응면역에서 중추적인 역할을 수행하는 대식세포의 활성화에 미치는 영향에 관하여 관찰하였다. 구절초에서 분리한 다당류를 마우스 유래 대식세포주인 RAW 264.7 cell에 처리하였을 때, iNOS의 발현을 조절하여 NO의 생성이 증가되었으며, 또한 면역활성 물질인 TNF-${\alpha}$, IL-$1{\beta}$ 및 IL-6 등의 cytokine의 분비능이 증가되었으며, 이러한 면역활성 매개자인 NO 및 cytokine의 증가의 원인에 관한 정확한 면역세포내 신호전달에 관하여 알아본 결과, CZPS 처리는 MAPKs(ERK, p38)의 인산화를 촉진시키며, 이로 인한 전사인자인 NF-${\kappa}B$의 인산화를 증가시켜, 대식세포의 활성화를 유도시키는 것으로 관찰되었다.

• Journal of Acupuncture Research
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• 제25권5호
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• pp.105-116
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• 2008

Objectives : The objective of this study is to investigate the suppressing effect of the cervi pantotrichum cornu pharmacopuncture on the expression of iNOS mRNA and production of NO in synoviocytes from artificially arthritis-induced mice. Methods : In vitro test, synoviocytes extracted from a knee joint of a mouse were cultivated, and the herbal extract of cervi pantotrichum cornu($0.4mg/m{\ell}$, $0.6mg/m{\ell}$, $0.8mg/m{\ell}$, and $1.0mg/m{\ell}$) was added into the wells of synoviocytes to suppress the expression of iNOS mRNA and production of NO. In vivo test, each ten mice were allocated into three groups; Normal group, CIA-elicitated group(CIA), and group treated with cervi pantotrichum cornu pharmacopuncture after CIA elicitation(CCA). The extract of cervi pantotrichum cornu was injected into the acupoint of $SP_{10}$ to observe the changes of foot thickness in mice and the suppression of MIF, TNF-$\alpha$, NF-${\kappa}B$ p65, and iNOS. Results : In vitro test, the expression of iNOS mRNA and production of NO were dose-dependently decreased in the wells of synoviocytes treated with PMA. In vivo test, the suppression of MIF, TNF-$\alpha$, NF-${\kappa}B$ p65, and iNOS was clearly shown in the pieces of the synovial joint treated with the extract of cervi pantotrichum cornu. The foot thickness also decreased dose-dependently. Conclusions : It is speculated that the cervi pantotrichum cornu pharmacopuncture can be applicable to the therapy of rheumatoid arthritis by suppressing the expression of iNOS mRNA and production of NO.

• 대한한방소아과학회지
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• 제32권4호
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• pp.71-86
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• 2018

Objectives The purpose of this study is to investigate the effects of Gardenia jasminoides for. grandiflora extracts' (GAJ) anti-inflammatory effect on RBL-2H3 mast cells and OVA/alum-sensitized mice. Methods In this study, IL-4 and IL-13 production was measured via ELISA analysis, and mRNA expressions of GM-CSF, IL-4, IL-5, $TNF-{\alpha}$, IL-6 were analyzed by real-time PCR. In addition, MAPKs and $NF-{\kappa}B$ p65 transcription factors were examined using western blotting, and ELISA was used to understand IgE, IL-4, and IL-13 production in ovalbumin-allergic mice in in vitro study. Results As a result of this study, 1. GAJ were observed to suppress the mRNA expression of GM-CSF, IL-4, IL-13, IL-5, $TNF-{\alpha}$, IL-6 in comparison to PMA 50 ng/ml, ionomycin $0.5{\mu}M$ (PI) control group. 2. GAJ also inhibited the IL-4, IL-13 production in comparison to PI control group. 3. Western blot analysis showed decrease on the expression of mast-cell-specific transcription factors, including MAKPs (ERK, JNK, p38) and $NF-{\kappa}B$ p65. 4. Orally-administered GAJ group in OVA/alum induced Balb/c mice showed decreased level of OVA-specific IgE in the serum. This group also has shown decreased the level of IL-4, IL-13 in the splenocyte culture supernatant. Conclusions Obtained results suggest that GAJ may regulate the allergic inflammation by transcription factors MAKPs (ERK, JNK, p38) and $NF-{\kappa}B$ p65 causing inhibition of Th2 cytokines in mast cells and OVA/alum-sensitized mice.

• 대한한의학회지
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• 제40권1호
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• pp.63-77
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• 2019

Objectives: The purpose of this study is to examine the effect of water and fermentation extracts of KMS (Kami-Mihudeongsikjang-tang) on AD (atopic dermatitis). Additionally, by applying the fermentation extracts of KMS at the first sensitization and latency period, I investgated whether it could prevent AD. Methods: In this study, to test the effect and preventive efficacy of KMSs on AD. DNCB-induced BALB/c mice of AD model was used. Through histological observation, epidermis and dermis thickness, the infiltration of eoshiphils and mast cells in epidermis and dermis were examined. We measured the serum level of IgE and IL-6, TNF-alpha, and the expressions of $NF-{\kappa}B$ and MAPK protein. In order to examine the effect of KMSs on keratinocyte, HaCaT cells were treated TNF-alpha and IFN-gamma to induce an inflammatory response. The KMSs were applied at various concentration in the experimental group. We investigated TARC expression. Results: The treatment groups were reduced epidermis and dermis thickness, inhibited the infiltration of eosinophils and mast cells, reduced the serum level of IL-6. Moreover, sfKMS group reduced serum level of TNF-alpha, inhibited the protein expressions of $NF-{\kappa}B$ and the phosphoryllation of ERK, JNK and p38. Especially sfKMS-pre group were reduced the serum level of IgE, show significant inhibition on the protein expression of $NF-{\kappa}B$ and the phosphoryllation of ERK, JNK and p38. In the experiment on HaCaT cells, sfKMS group were reduced expression of TARC. Conclusions: These result suggest that wKMS and sfKMS was effective in the treament on AD, and sfKMS would prevent AD.

• 대한본초학회지
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• 제32권2호
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• pp.77-85
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• 2017

Objectives : This study aimed to investigate the unknown mechanisms behind the anti- inflammatory activity of Hyeonto-dan(HT) 70% ethanol extract on LPS-stimulated RAW 264.7 cells. Methods : Cells were treated with Hyeonto-dan 1 h prior to addition of 200 ng/mL of LPS. Cell viability was measured by the MTS assay. Nitric oxide levels were determined by the Griess assay. $PGE_2$ were measured using EIA kit. Pro-inflammatory cytokine production was measured by the enzyme-linked immunosorbent assay (ELISA). The expression of COX-2, iNOS, and MAPKs was investigated by Western blot, qRT-PCR. $NF-{\kappa}B$/p65 localization and interaction of the TLR-4 receptor with LPS was examined by immunofluorescence assays. Results : Hyeonto-dan had no cytotoxicity at the measured concentration. Hyeonto-dan inhibited NO production and pro-inflammatory cytokines such as IL-6, $TNF-{\alpha}$, and PGE2 as well as the protein and mRNA expression of iNOS and COX-2. Moreover, Hyeonto-dan inhibited the interaction between LPS and TLR-4 in murine macrophages. It suppressed phosphorylation of extracellular signal-regulated kinase (ERK 1/2), c-jun N-terminal kinase (JNK 1/2) and p38. Finally, it inhibited translocation of $NF-{\kappa}B$ in response to competitive LPS. Conclusions : Based on the results of this study, Hyeonto-dan inhibited the binding of TLR-4 receptor to LPS and inhibited the phosphorylation of extracellular signaling pathway MAPKs. These inhibitory effects are thought that the amount of $NF-{\kappa}B$ delivered to the nucleus was decreased and the inflammatory reaction was prevented by decreasing the production of LPS-induced $PGE_2$, NO, IL-6 and $TNF-{\alpha}$.

• Molecular & Cellular Toxicology
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• 제4권2호
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• pp.106-112
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• 2008

Parkinson's disease (PD) progresses severely by a gradual loss of dopaminergic neurons in the substantia nigra (SN). Epidemiological studies showed that the incidences of PD were reduced by smoking of which the major component, nicotine might be neuroprotective. But the function of nicotine, which might suppress the incidences of PD, is still unknown. Fortunately, recently it was reported that a glial reaction and inflammatory processes might participate in a selective loss of dopaminergic neurons in the SN. The levels of tumour necrosis factor (TNF)-${\alpha}$ synthesised by astrocytes and microglia are elevated in striatum and cerebrospinal fluid (CSF) in PD. TNF-${\alpha}$ kills the cultured dopaminergic neurons through the apoptosis mechanism. TNF-${\alpha}$ release from glial cells may mediate progression of nigral degeneration in PD. Nicotine pretreatment considerably decreases microglial activation with significant reduction of TNF-${\alpha}$ mRNA expression and TNF-${\alpha}$ release induced by lipopholysaccharide (LPS) stimulation. Thus, this study was intended to explore the role of nicotine pretreatment to inhibit the expressions of TNF-${\alpha}$ mRNA in human fetal astrocytes (HFA) stimulated with IL-$1{\beta}$. The results are as follows: HFA were pretreated with 0.1, 1, and $10{\mu}g/mL$ of nicotine and then stimulated with IL-$1{\beta}$ (100 pg/mL) for 2h. The inhibitory effect of nicotine on expressions of TNF-${\alpha}$ mRNA in HFA with pretreated $0.1{\mu}g/mL$ of nicotine was first noted at 8hr, and the inhibitory effect was maximal at 12 h. The inhibitory effect at $1{\mu}g/mL$ of nicotine was inhibited maximal at 24 h. Cytotoxic effects of nicotine were noted above $10{\mu}g/mL$ of nicotine. Moreover, Nicotine at 0.1, 1 and $10{\mu}g/mL$concentrations significantly inhibited IL-$1{\beta}$-induced TF-${\kappa}B$ activation. Collectively, these results indicate that in activated HFA, nicotine may inhibit the expression of TNF-${\alpha}$ mRNA through the pathway which suppresses the NF-${\kappa}B$ activation. This study suggests that nicotine might be neuroprotective to dopaminergic neurons in the SN and reduce the incidences of PD.

• 대한한방내과학회지
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• 제33권3호
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• pp.272-283
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• 2012

Objectives : The purpose of this study was to investigate the mechanism of protective effect of Jinmu-tang (JMT, Zhenwu-tang) extract on $H_2O_2$-induced cell death in C6 glial cells. Methods : Cultured C6 glial cells of white mice were pretreated with JMT extract and exposed to $H_2O_2$ for inducing cell death. We measure the cell viability by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay and investigate the cell morphology using a light microscope after crystal violet (CV) staining. Reactive oxygen species (ROS) formation was analyzed using a flow cytometer and a fluorescent microscope after staining with 2'7'-dichlorofluorescein diacetate (DCF-DA). DNA fragmentation was analyzed using a flow cytometer after propidium iodide (PI) staining and nuclei morphology was investigated using a fluorescent microscope after 2-[4-amidinophenyl]-6-indo-lecarbamidine dihydrochloride (DAPI) staining. We analyzed expression of Bax, processing of procaspase-3 and poly (ADP-ribose) polymerase (PARP), and activation of nuclear factor-${\kappa}B$ (NF-${\kappa}B$) by western blot method. Tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$) secretion was analyzed using Quantikine kit. Results : We determined the elevated cell viability by JMT extract on $H_2O_2$-induced C6 glial cell death. ROS formation, DNA fragmentation, $I{\kappa}B{\alpha}$ phosphorylation, NF-${\kappa}B$ activation, and secretion of TNF-${\alpha}$ induced by $H_2O_2$ are inhibited by JMT extract pre-treatment. JMT extract inhibits Bax expression, processing of caspase-3 and PARP that are critical biochemical markers of apoptotic cell death. Conclusions : These results suggest that JMT extract has a protective effect on $H_2O_2$-induced C6 glial cell death in various pathways.

• Journal of Ginseng Research
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• 제38권1호
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• pp.8-15
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• 2014

Helicobacter pylori-induced gastric inflammation includes induction of inflammatory mediators interleukin (IL)-8 and inducible nitric oxide synthase (iNOS), which are mediated by oxidant-sensitive transcription factor NF-${\kappa}B$. High levels of lipid peroxide (LPO) and increased activity of myeloperoxidase (MPO), a biomarker of neutrophil infiltration, are observed in H. pylori-infected gastric mucosa. Panax ginseng Meyer, a Korean herb medicine, is widely used in Asian countries for its biological activities including anti-inflammatory efficacy. The present study aims to investigate whether Korean Red Ginseng extract (RGE) inhibits H. pylori-induced gastric inflammation in Mongolian gerbils. One wk after intragastric inoculation with H. pylori, Mongolian gerbils were fed with either the control diet or the diet containing RGE (200 mg RGE/gerbil) for 6 wk. The following were determined in gastric mucosa: the number of viable H. pylori in stomach; MPO activity; LPO level; mRNA and protein levels of keratinocyte chemoattractant factor (KC, a rodent IL-8 homolog), IL-$1{\beta}$, and iNOS; protein level of phospho-$I{\kappa}B{\alpha}$(which reflects the activation of NF-${\kappa}B$); and histology. As a result, RGE suppressed H. pylori-induced mRNA and protein levels of KC, IL-$1{\beta}$, and iNOS in gastric mucosa. RGE also inhibited H. pylori-induced phosphorylation of $I{\kappa}B{\alpha}$ and increases in LPO level and MPO activity of gastric mucosa. RGE did not affect viable H. pylori colonization in the stomach, but improved the histological grade of infiltration of poly-morphonuclear neutrophils, intestinal metaplasia, and hyperplasia. In conclusion, RGE inhibits H. pyloriinduced gastric inflammation by suppressing induction of inflammatory mediators (KC, IL-$1{\beta}$, iNOS), MPO activity, and LPO level in H. pylori-infected gastric mucosa.

• 생명과학회지
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• 제22권11호
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• pp.1571-1586
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• 2012

지구상에는 약 5,400여 종의 포유동물이 있고 그 중 약 1,000여 종은 풀을 뜯어 먹고 사는 초식동물이다. 초식동물 중에서 약 250여 종이 반추동물로 알려져 있다. 반추동물인 소와 양은 반추위에서 주로 발효가 일어나지만 비반추동물인 돼지와 사람은 맹장과 결장, 직장에서 주로 발효가 일어난다. 반추위 미생물의 종류와 우점도 Bacteroidetes 51%, Firmicutes 43% 존재하며, 사람의 대장미생물의 우점도Firmicutes 65%, Bacteroidetes 25%로 존재한다. 풀의 세포벽 구성성분은 미생물에 의해 분해, 발효에 의해 SCFA (short chain fatty acid)를 생성하게 되고 acetate, propionate, butyrate 생성비율은 60:25:15이다. 장내 primary butyrate transporter인 MCT1(monocarboxylatetransports-1)에 의해서 흡수된 butyrate는 SCFA receptor GPR43과 GPR41을 활성화시킨다. Butyrate는 강력한 anti-tumorigenic 기능을 가지고 있다. Butyrate는 다양한 cancer cell에 효과를 나타내며 세포내의 기능 조절에 기여하고, 암세포사멸을 유도하는 특성이 있다. Butyrate는 caspase의 활성화, HDAC (histone deacetylase) 활성을 억제하여apoptosis를 유도하고, p53 발현증가로 cell cycle arrest와 apoptosis를 유도한다. SCFA의 항 염증작용으로는 장 상피세포에서 IL-8 발현 감소, NO합성과 NF-${\kappa}B$ (nuclear factor ${\kappa}B$)의 활성을 억제하여 염증으로 인한 암 발생을 억제한다. Butyrate는 장 점막의 생리적 기능을 유지하는데 중요한 역할을 하며 IBD (inflammatory bowel disease) 치료법으로 이용되고 있다.