• 대한한방부인과학회지
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• 제26권4호
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• pp.66-81
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• 2013

Purpose: This study was carried out to investigate the anti-inflammatory effects of Ligustri Lucidi Fructus water extract (LF) in the lipopolysaccharide (LPS)-induced mouse macrophages RAW 264.7 cell. Methods: Ligustri Lucidi Fructus was extracted with distilled water (2,000 ml) for 2 hours. In order to evaluate cytotoxicity of LF, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was performed. To investigate anti-inflammatory effects of LF, the concentration of nitric oxide (NO) was measured with NO assay, cytokine was measured by Bio-Plex cytokine assay, and intracellular calcium (Ca) was measured with Fluo-4 Ca assay in RAW 264.7 cell. And when p-value is below 0.05, it is judged to have the significant difference statistically (P<0.05). Results: 1. LF showed no cytotoxicity. 2. LF inhibited significantly the production of NO at the concentration of 25, 50 and $100{\mu}g/ml$. 3. LF inhibited significantly the production of interleukin (IL)-4, macrophage inflammatory protein (MIP)-$1{\alpha}$, granulocyte colony stimulating factor (G-CSF) at the concentration of 25, 50, 100 and $200{\mu}g/ml$. 4. LF inhibited significantly the production of granulocyte macrophage-colony stimulating factor (GM-CSF), vascular endothelial growth factor (VEGF) at the concentration of 50, 100 and $200{\mu}g/ml$, the interferon (IFN)-${\gamma}$ at 25, 50 and $100{\mu}g/ml$ respectively. 5. LF inhibited significantly the production of IL-$1{\beta}$ at the concentration of 50 and $200{\mu}g/ml$, the IL-5 at 25 and $100{\mu}g/ml$, the IL-12p70, MIP-$1{\beta}$ at 50 and $100{\mu}g/ml$, the regulated on activation, normal T cell expressed and secrete d (RANTES) at 100 and $200{\mu}g/ml$ respectively. 6. LF inhibited significantly the production of IL-10, interferon gamma-induced protein (IP)-10 at the concentration of $200{\mu}g/ml$. 7. LF inhibited significantly the production of intracellular Ca at the concentration of 25, 50, 100 and $200{\mu}g/ml$. Conclusions: These results suggest that LF has anti-inflammatory effect and immuno-modulating activity.

• Journal of Ginseng Research
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• 제39권3호
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• pp.230-237
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• 2015

Background: Colorectal cancer (CRC) is a leading cause of death worldwide. Chronic gut inflammation is recognized as a risk factor for tumor development, including CRC. American ginseng is a very commonly used ginseng species in the West. Methods: A genetically engineered $Apc^{Min/+}$ mouse model was used in this study. We analyzed the saponin composition of American ginseng used in this project, and evaluated its effects on the progression of high-fat-diet-enhanced CRC carcinogenesis. Results: After oral ginseng administration (10-20 mg/kg/d for up to 32 wk), experimental data showed that, compared with the untreated mice, ginseng very significantly reduced tumor initiation and progression in both the small intestine (including the proximal end, middle end, and distal end) and the colon (all p < 0.01). This tumor number reduction was more obvious in those mice treated with a low dose of ginseng. The tumor multiplicity data were supported by body weight changes and gut tissue histology examinations. In addition, quantitative real-time polymerase chain reaction analysis showed that compared with the untreated group, ginseng very significantly reduced the gene expression of inflammatory cytokines, including interleukin-$1{\alpha}$ (IL-$1{\alpha}$), IL-$1{\beta}$, IL-6, tumor necrosis factor-${\alpha}$, granulocyte-colony stimulating factor, and granulocyte-macrophage colony-stimulating factor in both the small intestine and the colon (all p < 0.01). Conclusion: Further studies are needed to link our observed effects to the actions of the gut microbiome in converting the parent ginsenosides to bioactive ginseng metabolites. Our data suggest that American ginseng may have potential value in CRC chemoprevention.

• 한방재활의학과학회지
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• 제24권3호
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• pp.71-85
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• 2014

Objectives In this study, we investigated the inhibitory effects of Samhwangsasim-tang (S.H) on the allergic response caused by ovalbumin(OVA) sensitization and challenge in BALB/c mice. Methods The experimental animals were divided into five groups; 1) normal as negative control, 2) OVA-sensitized mice, 3) OVA-sensitized mice treated with 200 mg/kg of S.H 200, 4) OVA-sensitized mice treated with 400 mg/kg of S.H 400, and 5) OVA-sensitized mice treated with 5 mg/kg of Dexamethasone (Dex). Antigen sensitization for allergic mouse model was performed with twice injection of OVA for 2 weeks. After secondary injection, S.H was administrated orally into mice every day for 13 days and the inhibitory effect of S.H on allergic responses was evaluated. Results Treatment of S.H into allergic mice reduced significantly ear edema and infiltration of immune cells in ear tissues induced with OVA challenge in a dose-dependent manner. S.H reduced significantly the serum levels of Total Immunoglobulin(Ig)G and IgE, and particularly inhibited the production of OVA-specific IgE, but not OVA-specific IgG. The serum level of proinflammatory cytokine TNF-${\alpha}$ and Th2-associated cytokine IL-4 also were significantly decreased by S.H adminstration in a dose denpendent manner. S.H attenuated OVA-induced secretion of IFN-${\gamma}$, but not IL-12 which is a cytokine inducing the development of Th1 cells. It also reduced significantly the secretion of IL-4, which is a cytokine inducing the development of Th2 cells, after splenocytes were stimulated with OVA. However the secretion of IL-5 and IL-13 was influenced weakly or a little. Conclusions These results indicate that S.H could reduce the allergic response through inhibition of antigen-specific IgE and Th2-inducing cytokines. It suggest that S.H may be available clinically for the treatment of allergic patients.

• 한국식품영양과학회지
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• 제45권4호
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• pp.613-618
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• 2016

1,2,3,4,6-Penta-O-galloyl-${\beta}$-D-glucose(PGG)는 오배자(Galla Rhois)의 gallotannin으로 항산화 효과, 항균 효과, 항알레르기 효과 등을 가지는 것으로 알려졌다. 알레르기성질환은 비만세포를 포함한 다양한 면역세포와 관련된 질환이다. 비만세포는 탈과립화에 의한 알레르기 매개성 물질(histamine 및 ${\beta}-hexosaminidase$)의 분비, T helper 2 type 사이토카인(IL-4) 분비 및 염증을 일으키는 매개체($TNF-{\alpha}$)의 증가를 통해 알레르기 반응에서 중요한 역할을 한다. 따라서 본 연구에서는 PGG가 phorbol 12-myristate 13-acetate(PMA)와 A23187로 비만세포인 RBL-2H3 세포에서 탈과립 그리고 Th2 type 및 염증성 사이토카인 생산에 미치는 영향을 조사하고, 관련 메커니즘에 대해 평가하였다. PGG는 PMA와 A23187 자극에 의한 탈과립 마커(histamine 및 ${\beta}-hexosaminidase$)의 분비를 억제하고, IL-4 및 $TNF-{\alpha}$와 같은 사이토카인의 분비를 억제하였다. 이러한 효과는 전사인자인 $NF-{\kappa}B$의 세포질에서 핵으로의 이동을 억제함으로써 나타나는 것으로 판단된다. 이러한 결과로부터 gallotannin의 하나인 PGG가 알레르기 반응을 현저히 저해하는 효과가 있는 것으로 나타나, 향후 알레르기성 질환을 예방, 개선 및 치료하는 데 유용한 물질로 사용될 가능성이 있을 것으로 생각된다.

• 한국식품영양과학회지
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• 제44권2호
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• pp.182-190
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• 2015

본 연구는 홍삼추출물(홍삼단)과 생약복합추출물(헤모힘)이 주요 기능성분은 다르지만 면역활성이라는 생리활성 측면에서 동일한 효과를 인정하고 있어서 이들의 병용 처리가 면역세포에 어떠한 영향을 미치는지에 대해 알아보기 위하여 수행되었다. 면역활성능에 관한 평가를 진행하기 위하여 마우스의 골수에서 분리한 미분화 골수세포를 선천면역에서 중요한 역할을 수행하는 대식세포 및 수지상세포로 분화시킨 후 홍삼추출물과 생약복합추출물을 병용 처리하였을 때 대식세포 및 수지상세포의 세포 증식능 및 사이토카인의 분비능이 증가되는 것으로 관찰되었다. 또한 활성화된 탐식(면역)세포의 세포 표면에서 발현되는 CD80과 CD86의 발현과 탐식(면역)세포의 항원제시에 밀접한 관련이 있는 주 조직적합성 복합체(MHC class II)의 발현이 홍삼추출물과 생약복합추출물의 병용 처리구에서 유의적으로 증가되는 것으로 관찰되었다. 또한 후천면역에서 중요한 역할을 수행하는 면역 T 세포가 다량으로 분포하는 비장 조직으로부터 비장세포를 분리하여 홍삼추출물과 생약복합추출물을 병용처리하였을 때 세포 증식능 및 면역활성을 유도하는 Th1 세포가 분비하는 사이토카인의 함량이 증가되는 것으로 나타났다. 이러한 결과로 미루어 볼 때 홍삼추출물과 생약복합 추출물의 병용 처리는 선천면역뿐만 아니라 후천면역에 관여하는 다양한 면역세포의 활성화에 직 간접적으로 다양한 상승작용을 미치는 것으로 사료된다.

• 대한약침학회지
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• 제20권2호
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• pp.127-131
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• 2017

Objectives: Polymorphonuclear neutrophils (PMNs) constitute the first line of defense against invading microbial pathogens. Early events in inflammation involve the recruitment of neutrophils to the site of injury or damage where changes in intracellular calcium can cause the activation of pro-inflammatory mediators from neutrophils including superoxide generation, degranulation and release of myeloperoxidase (MPO), productions of interleukin (IL)-8 and tumor necrosis factor ${\alpha}$ ($TNF-{\alpha}$), and adhesion to the vascular endothelium. To address the anti-inflammatory role of flavonoids, in the present study, we investigated the effects of the flavonoids quercetin and vitexin on the stimulus-induced nitric oxide (NO), $TNF-{\alpha}$, and MPO productions in human neutrophils. Methods: Human peripheral blood neutrophils were isolated, and their viabilities were determined by using the Trypan Blue exclusion test. The polymorphonuclear leukocyte (PMNL) preparations contained more than 98% neutrophils as determined by morphological examination with Giemsa staining. The viabilities of cultured neutrophils with various concentrations of quercetin and vitexin ($1-100{\mu}M$) were studied using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assays. Neutrophils were cultured in complete Roswell Park Memorial Institute (RPMI) medium, pre-incubated with or without quercetin and vitexin ($25{\mu}M$) for 45 min, and stimulated with phorbol 12-myristate 13-acetate (PMA) ($10^{-7}M$). NO production was carried out through nitrite determination by using the Griess method. Also, the $TNF-{\alpha}$ and the MPO productions were measured using enzyme-linked immunosorbent assay (ELISA) kits and MPO assay kits. Results: Neutrophil viability was not affected up to a concentration of $100{\mu}M$ of quercetin or vitexin. Both quercetin and vitexin significantly inhibited $TNF-{\alpha}$, NO, and MPO productions in human neutrophils (P < 0.001). Conclusion:The present study showed that both quercetin and vitexin had significant anti-inflammatory effects. Thus, treatment with either quercetin or vitexin may be considered as a therapeutic strategy for treating patients with neutrophil-mediated inflammatory diseases.

• 대한약침학회지
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• 제20권2호
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• pp.93-99
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• 2017

Objectives: Selaginella bryopteris L. (family: Selaginaceae), is often used in traditional Indian systems of medicine for the prevention and cure of several disorders and for the treatment of patient with spermatorrhoea, venereal disease, constipation, colitis, urinary tract infections, fever, epilepsy, leucorrhoea, beri-beri and cancer. It is also used as a strength tonic. This study aimed to evaluate the mechanisms underlying the anti-inflammatory effects of topically administered aqueous, polar and non-polar methanolic fractions ($10mg/20{\mu}L$) of Selaginella bryopteris. Methods: An acute oral toxicity study of Selaginella bryopteris at doses from 250 to 2,000 mg/kg body weight (bw) was performed. Aqueous, polar and non-polar methanolic extracts ($10mg/20{\mu}L$) applied topically for 5 days were evaluated for their anti-inflammatory effects against 12-tetra-O-decanoyl phorbol acetate (TPA)- induced inflammation by using the redness in the ear, the ear's weight (edema), oxidative stress parameters, such as lipid-peroxide (LPO) and nitric oxide (NO), and the pro-inflammatory cytokines involved in inflammation, such as tumour necrosis factor $(TNF)-{\alpha}$, interleukin $(IL)-1{\beta}$ and IL-6. Indomethacine ($0.5mg/20{\mu}L$) was used for the positive control. Results: Selaginella bryopteris produced no mortalities when administered orally at doses from 250 to 2,000 mg/kg bw. Topical treatment with the non-polar methanolic fraction ($10mg/20{\mu}L$) significantly suppressed redness ($2.4{\pm}0.5$) and edema ($30.4{\pm}1$) and effectively reduced the LPO level ($32.3{\pm}3.3$). The NO level was ($8.07{\pm}0.55$), and the $TNF-{\alpha}$, $IL-1{\beta}$, and IL-6 levels were decreased to $69.6{\pm}15.5$, $7.7{\pm}4.8$ and $82.6{\pm}5.9$, respectively. Conclusion: This study demonstrated for the first time the mechanisms underlying the anti-inflammatory effect of medicinal plants like Selaginella bryopteris and quantified the pharmacological interactions between them. The present study showed this herbal product to be a promising anti-inflammatory phytomedicine for the treatment of patients with inflammatory skin diseases.

• 대한본초학회지
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• 제25권4호
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• pp.11-16
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• 2010

Objective : Oriental medicines have been combined oriental medical theory which based on the seven modes of emotions. Notopterygium incisum (N. incisum) and Clematis manshurica (C. manshurica) have been used as an anti-rheumatic and analgesic medicine for the treatment of rheumatism, headache, cold, etc. In this study, we evaluate the synergistic anti-inflammatory effect of N. incisum and C. manshurica. Method : To evaluate the synergistic anti-inflammatory effect of a herbal mixture N. incisum and C. manshurica, we examined the changed ear thickness in 12-O-tetradecanoyl-phorbol-13-acetate (TPA)-induced mouse ear edema model after topical application of herbal mixture. In addition, the levels of markers for inflammation, such as tumor necrosis factor (TNF)-${\alpha}$, interleukin (IL)-$1{\beta}$, prostaglandin $G_2$ ($PGE_2$), and nitric oxide (NO) were measured by ELISA assay and Griess reagent in lipopolysaccharide-stimulated Raw 264.7 cells. Results : Our results showed that aqueous extracts of N. incisum and C. manshurica combination significantly inhibited the mouse ear edema induced by TPA. Moreover, the aqueous extracts of N. incisum and C. manshurica combination exhibited synergistic effects in down-regulating TNF-${\alpha}$, IL-$1{\beta}$, $PGE_2$ level, but not NO. Conclusions : This study suggested that combined treatment of N. incisum and C. manshurica, based on seven methods in prescription compatibility, has a synergistic effect in down-regulating inflammatory response both in vitro and in vivo models.

• Journal of Microbiology and Biotechnology
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• 제18권9호
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• pp.1606-1612
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• 2008

Interleukin (IL)-32 is a recently identified proinflammatory cytokine that is one of the IL-18 inducible genes, and plays an important role in autoimmune and inflammatory diseases. We produced antibodies against IL-32 and studied the expression of IL-32 in human stomach cancer. We detected IL-32 secreted from K-562 cells which were stably transfected with IL-32 and in the sera of stomach cancer patients by a sandwich ELISA using a monoclonal antibody KU32-52 and a polyclonal antibody. In order to optimize a sandwich immunoassay, recombinant IL-32a was added, followed by the addition of a biotinylated KU32-52 into microtiter plate wells precoated with a goat anti-IL-32 antibody. The bound biotinylated KU32-52 was probed with a streptavidin conjugated to HRP. This sandwich ELISA was highly specific and had a minimal detection limit of 80 pg/ml (mean${\pm}$SD of zero calibrator) and measuring up to 3,000 pg/ml. This ELISA showed no cross-reaction with other cytokines such as hIL-1$\alpha$, hIL-1$\beta$, hIL-2, hIL-6, hIL-8, hIL-10, hIL-18, and hTNF-$\alpha$. Intra-assay coefficients of variation were 18.5% to 4.6% (n=10), and inter-assay coefficients were 23% to 9% (n=10). The average IL-32 level in the sera of 16 stomach cancer patients (189 pg/ml) was higher than that of 12 healthy control men (109 pg/ml). Our results indicate that serum IL-32 level can be detected by using an established ELISA, and that this immunoassay and mAb KU32-09 specific for immunohistochemistry can be used in the detection of expressed and secreted IL-32 in stomach cancer patients.

• Nutrition Research and Practice
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• 제5권3호
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• pp.219-223
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• 2011

Obesity has been reported to be associated with low grade inflammatory status. In this study, we investigated the inflammatory response as well as associated signaling molecules in immune cells from diet-induced obese mice. Four-week-old C57BL mice were fed diets containing 5% fat (control) or 20% fat and 1% cholesterol (HFD) for 24 weeks. Splenocytes ($1{\times}10^7$ cells) were stimulated with $10\;{\mu}g/mL$ of lipopolysaccharide (LPS) for 6 or 24 hrs. Production of interleukin (IL)-$1{\beta}$, IL-6, and TNF-${\alpha}$ as well as protein expression levels of nucleotide-binding oligomerization domain (NOD)2, signal transducer and activator of transcription (STAT)3, and pSTAT3 were determined. Mice fed HFD gained significantly more body weight compared to mice fed control diet ($28.2{\pm}0.6$ g in HFD and $15.4{\pm}0.8$ g in control). After stimulation with LPS for 6 hrs, production of IL-$1{\beta}$ was significantly higher (P=0.001) and production of tumor necrosis factor (TNF)-${\alpha}$ tended to be higher (P < 0.064) in the HFD group. After 24 hrs of LPS stimulation, splenocytes from the HFD group produced significantly higher levels of IL-6 ($10.02{\pm}0.66$ ng/mL in HFD and $7.33{\pm}0.56$ ng/mL in control, P=0.005) and IL-$1{\beta}$ ($121.34{\pm}12.72$ pg/mL in HFD and $49.74{\pm}6.58$ pg/mL in control, P < 0.001). There were no significant differences in the expression levels of STAT3 and pSTAT3 between the HFD and the control groups. However, the expression level of NOD2 protein as determined by Western blot analysis was 60% higher in the HFD group compared with the control group. NOD2 contributes to the induction of inflammation by activation of nuclear factor ${\kappa}B$. These findings suggest that diet-induced obesity is associated with increased inflammatory response of immune cells, and higher expression of NOD2 may contribute to these changes.