• 동의생리병리학회지
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• 제33권2호
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• pp.131-140
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• 2019

The objective of present study is to evaluate concentration-dependent in vitro anti-osteoarthritic (OA) effects of synergic mixed formula consisted of dried pomegranate juice concentrate powder, Eucommiae Cortex aqueous extract and Achyranthis Radix aqueous extract 5:4:1 (g/g) mixture on the primary cultured rat articular chondrocytes. First, any cytotoxic effect of mixture was observed using MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium Bromide) assay. Next, cyto-protective effect of test substances was evaluated by using the recombinant human interleukin $(rhIL)-1{\alpha}$ induced chondrocytes. In addition, anti-inflammatory effects were also observed on the lipopolysaccaride (LPS) treated chondrocytes through prostaglandin $E_2(PGE_2)$ productions and 5-lipoxygenase (LPO) activities, and inhibitory effects on matrix metalloproteinase (MMP)-2 and MMP-9 activities were observed on $rhIL-1{\alpha}$ treated chondrocytes with their extracellular matrix (ECM) related mRNA expressions. No obvious cytotoxic effects of mixture were demonstrated. Inflammatory damages of chondrocytes and related ECM degradations induced by treatment of LPS or $rhIL-1{\alpha}$ were significantly and concentration-dependently inhibited by pretreatment of mixture from a concentration level of 0.001 mg/ml to 1 mg/ml. In addition, mixture showed $IC_{50}$ for $rhIL-1{\alpha}-induced$ MMP-2 and MMP-9 activities as 44.01 and $162.47{\mu}g/ml$, and also showed $EC_{50}$ for $rhIL-1{\alpha}-induced$ inhibition of collagen type II, SOX9 and aggrecan mRNA expression as 8.61, 10.79 and $4.47{\mu}g/ml$, respectively. It is observed that mixture showed concentration-dependent anti-inflammatory and cytoprotective ECM preserved effects on the primary cultured rat articular chondrocytes without cytotoxicity.

• Korean Journal of Acupuncture
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• 제39권2호
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• pp.43-53
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• 2022

Objectives : Paeonia lactiflora Pallas (PLP) have been reported to have pharmacological effects such as anti-inflammatory and analgesic. However, it is not yet known whether PLP extract has anti-inflammatory effect on HaCaT cells, human keratinocyte. Methods : To confirm the anti-inflammatory effect of PLP on keratinocyte, TNF-𝛼/IFN-𝛾-stimulated HaCaT cells were used. HaCaT cells were pre-treated with PLP for 1h before stimulation with TNF-𝛼/IFN-𝛾. Then HaCaT cells were stimulated with TNF-𝛼/IFN-𝛾 for 24 h, the cells and media were harvested to measure the inflammatory cytokines levels. Granulocyte-macrophage colony stimulating factor (GM-CSF), monocyte chemoattractant protein-1 (MCP-1), interleukin 1 beta (IL-1𝛽), and TNF-𝛼 were analyzed by enzyme-linked immunosorbent assay (ELISA), and the mRNA expression of thymus and activation-regulated chemokines (TARC), IL-6, and IL-8 were measured by reverse transcription-polymerase chain reaction (RT-PCR). We also investigated the inhibitory mechanism of the mitogen-activated protein kinase (MAPKs) including ERK, JNK, and p38 and nuclear factor-kappaB (NF-𝜅B) by PLP using western blot. Results : PLP did not show cytotoxicity in HaCaT cells. In TNF-𝛼/IFN-𝛾-stimulated HaCaT cells, PLP significantly inhibited the expression of GM-CSF, MCP-1 IL-1𝛽, TNF-𝛼, TARC and IL-6. PLP inhibited the phosphorylation of ERK and translocation of NF-𝜅B into the nucleus. Conclusions : These results indicate that PLP could ameliorate the TNF-𝛼/IFN-𝛾-stimulated inflammatory response through inhibition of MAPK and NF-kB signal pathway. This suggests that PLP could be used beneficial agent to improve skin inflammation.

• Asian-Australasian Journal of Animal Sciences
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• 제29권2호
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• pp.250-256
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• 2016

The objective of this study was to investigate the effects of graded levels of isomaltooligosaccharides (IMO) on the performance, immune function and intestinal microflora and intestinal mucosal morphology of weaned pigs. In a 28-day experiment, one hundred eighty, twenty eight-day-old, crossbred (Duroc${\times}$Large White${\times}$Landrace), weaned pigs, with an initial body weight of $8.19{\pm}1.45kg$, were fed either an unsupplemented corn-soybean meal based diet or similar diets supplemented with 0.2%, 0.4%, 0.6%, or 0.8% IMO added at the expense of corn. Each treatment was replicated six times with six pigs (three barrows and three gilts) per pen. From day 0 to 14, weight gain was linearly increased (p<0.05), while gain:feed (p<0.05) was linearly improved and diarrhea rate (p = 0.05) linearly declined as the IMO level increased. On d 14, the level of the immunoglobulins IgA, IgM, and IgG in the serum of pigs were linearly increased (p<0.05) with increasing IMO supplementation. Interleukin-6 (IL-6) was linearly (p<0.05) and quadratically (p<0.05) decreased as IMO intake increased. From day 15 to 28, there was a trend for weight gain to be linearly increased, and IL-2 was linearly (p<0.05) increased as IMO supplementation increased on d 28. Over the entire experiment, weight gain was linearly increased (p<0.05), while gain:feed (p<0.05) was linearly improved and diarrhea rate (p<0.05) was linearly decreased as the IMO level increased. Supplementation with IMO had no effect on the intestinal microflora of pigs in the ileum and cecum of pigs, as well as the villus height and crypt depth in the ileum and jejunum (p>0.05). These results indicate that dietary inclusion of IMO increases weight gain, gain:feed and enhanced the immune status of pigs, and could be a valuable feed additive for use in weaned pigs, particularly during the period immediately after weaning.

• Journal of Korean Neurosurgical Society
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• 제38권2호
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• pp.126-131
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• 2005

Objective : The choice of tumor antigen for dendritic cell[DC]-loading has still been an unresolved problem in the DC-based vaccine strategies against malignant gliomas that has not been found well-characterized tumor specific antigens. In this study, we compare tumor-specific T cell response induced by glioma apoptotic body[GAB]-pulsed DCs to response induced by glioma cell lysate-pulsed ones quantitatively. Methods : DCs generated in the presence of granulocyte macrophage-colony stimulating factor and interleukin[IL]-4 from peripheral blood mononuclear cells[PBMCs] of HLA-A2 positive healthy donors were cultured. Each GABs and glioma cell lysate generated from HLA-A2 positive T98G glioblastoma cells were co-incubated with DCs. $CD8^+$ T lymphocytes isolated from PBMCs of same donors were cultured in media containing IL-2 and either stimulated by GAB- or lysate-pulsed DCs three times at a weekly interval. The interferon[IFN]-${\gamma}$ concentrations of each cell culture supernate were measured by enzyme immunoassay technique. Cytolytic activity of the generated cytotoxic $CD8^+$ T cells either stimulated with GAB- or lysate-pulsed DCs was determined by a standard 4-h $^{51}Cr$-release assay. Results : IFN-${\gamma}$ production and cytolytic activity of effector T cells stimulated by GAB-pulsed DCs were significantly higher than those of T cells stimulated by lysate-pulsed ones. Conclusion : These results indicate the choice of antigen is a critical determinant in the induction of antitumor immunity against malignant glioma. Antigen preparations from GABs represent a promising alternative to glioma cell lysate in DC-based glioma vaccine strategies.

• 생명과학회지
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• 제19권10호
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• pp.1352-1359
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• 2009

본 연구는 장시간의 고강도 트레드밀 운동이 호흡기계의 생리를 가장 잘 반영하는 기관지 폐포계에 미치는 생리학적 효과를 규명하기 위해 실시되었다. 랫트를 대상으로 매일 8주간 운동시킨 결과 60분 및 120분 운동 그룹군은 대조군(비운동군)과 30분 운동군보다 체중이 유의하게 낮았으며 운동전보다 체중증가율도 높지 않았다. 기관지 폐포세척액 내 총 백혈구수는 세 운동군(30분, 60분, 120분) 모두 대조군보다 유의하게 높았다. 호중구수는 대조군에 비해 30분군과 120분군은 유의하게 증가하였고 60분군은 감소하였으며 30분군이 가장 높았다. 림프구수는 세 운동군 모두 대조군에 비해 유의하게 높았고 120분군이 가장 높았다. 대식세포(단구) 수는 세군 모두 대조군보다 높았고 120군이 가장 높았다. 기관지폐포액 내 인터루킨-6 농도는 대조군보다 30분군은 낮았고 60분과 120분군은 높았다. 그러나 인터루킨-10과 인터페론 감마 농도는 세 운동군 모두 대조군에 비해 유의하게 낮았다. 산화질소(nitric oxide) 농도는 세 운동군 모두 대조군보다 유의하게 높았다. 본 연구의 결과들은 1시간 이상의 고강도 트레드밀 운동은 기관지 폐포 내 백혈구 분포도에 변화를 유도하고 염증성 사이토카인 및 산화성질소 농도의 증가와 항염증성 사이토카인 농도의 감소와 같은 유해한 효과를 일으킬 수 있음을 시사하고 있다.

• 한국식품영양과학회지
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• 제39권8호
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• pp.1213-1219
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• 2010

본 연구는 흑마늘을 기능성 화장품 소재로 활용하기 위하여 그 기능성을 in vitro에서 tyrosinase 및 elastase 저해 활성을 측정한 결과 피부노화에 관여하는 두 효소의 활성을 모두 저해하였다. 흑마늘 추출물을 hartley계 기니픽 수컷을 사용하여 피부 1차 자극 실험을 실시한 결과 1차 피부자극지수(P.I.I.)가 0.23으로 practically non-irritation(비자극성)에 해당하는 자극으로 피부 자극이 거의 없었음을 알 수 있었다. Maximization test법으로 피부 감작성을 확인한 결과 시험물질에 의한 홍반과 부종 등이 전혀 유발되지 않았다. 이상의 결과로부터 흑마늘 추출물에 대한 기니픽의 피부 감작율은 0%로, 피부에 대한 피부 감작성이 없는 것으로 확인되었다. 따라서 이러한 본 연구의 결과는 흑마늘 추출물이 기능성 화장품 소재로 피부노화를 억제하는 기능성과 안전성확보된 소재임을 추정할 수 있었다.

• 한국식품영양과학회지
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• 제45권8호
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• pp.1099-1106
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• 2016

본 연구는 꾸지뽕 잎으로부터 추출된 다당류인 CTP의 처리가 면역세포의 활성에 미치는 영향에 관하여 평가하였다. CTP는 에탄올 침전법에 의하여 추출하였고, 면역 활성능의 평가는 대식세포주인 RAW 264.7 세포와 미분화 골수세포로부터 유도 분화시킨 대식세포 및 마우스 비장으로부터 유리시킨 비장세포에 CTP를 농도별로 처리하여 관찰하였다. 선천면역계에서 중요한 역할을 수행하는 대식세포에 CTP를 처리하였을 때 세포 증식률, NO 및 cytokine 분비능이 CTP 처리 농도 의존적으로 증가하는 것으로 관찰되었을 뿐만 아니라 비장세포에서도 이와 유사하게 세포 증식률이 증가하고 Th 1 type의 cytokine 분비능 또한 유의적으로 증가하는 것으로 관찰되었다. 이상의 결과로 미루어보아 꾸지뽕다당류 추출물인 CTP는 다양한 면역세포의 활성을 증가시키는 것으로 생각하며 이를 활용하여 다양한 식품 및 건강보조식품을 개발한다면 그 경제적 가치가 매우 클 것으로 생각한다.

• BMB Reports
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• 제47권2호
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• pp.115-120
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• 2014

In this study, we show that Mycobacterium avium subsp. paratuberculosis MAP1305 induces the maturation of bone marrow-derived dendritic cells (BMDCs), a representative antigen presenting cell (APC). MAP1305 protein induces DC maturation and the production of pro-inflammatory cytokines (Interleukin (IL)-6), tumor necrosis factor (TNF)-${\alpha}$, and IL-$1{\beta}$) through Toll like receptor-4 (TLR-4) signaling by directly binding with TLR4. MAP1305 activates the phosphorylation of MAPKs, such as ERK, p38MAPK, and JNK, which is essential for DC maturation. Furthermore, MAP1305-treated DCs transform naive T cells to polarized $CD4^+$ and $CD8^+$ T cells, thus indicating a key role for this protein in the Th1 polarization of the resulting immune response. Taken together, M. avium subsp. paratuberculosis MAP1305 is important for the regulation of innate immune response through DC-mediated proliferation of $CD4^+$ and $CD8^+$ T cells.

• 한방재활의학과학회지
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• 제21권4호
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• pp.49-65
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• 2011

Objectives: This study was performed to investigate the effects of Gagamsosokmyeong-tang(Jiajianxiaoxuming-tang) on the monosodium iodoacetate(MIA) induced early state osteoarthritis in rats. Methods: Osteoarthritis was induced by injection of MIA(0.25 mg) into knee joints of rats. Osteoarthritis rats were divided into control(n=8) and treated=8 group respectively, Control group was taken distilled water and treated group was taken extracts of Gagamsosokmyeong-tang(Jiajianxiaoxuming-tang) by orally for 20 days. Body weight was measured at 0, 5, 10, 15, 20 days after MIA injection. At the end of experiment, gross and histopathological examination on the articular cartilages of the knee joints were performed. Proteoglycan(PG) content of articular cartilages were analysed by safranine O staining method. The content of tumor necrosis $factor-{\alpha}(TNF-{\alpha})$, $interleukin-1{\beta}(IL-1{\beta})$ in synovial fluids were analysed by enzyme-inked inmunosorbent assay(ELISA) method. And also cycloxygenase-2(COX-2), matrix metalloproteinase 3(MMP-3), inducible nitric oxide synthase(iNOS), calpain immunochistochemical examination on the knee joints were performed. Results: PG content in articular cartilages of the treated group was significantly increased compared with control group. Histopathological osteoarthritic score of the treated group was significantly decreased compared with control group. $TNF-{\alpha}$ content in synovial fluids, expression of iNOS and calpain in synovial membrane of the treated group were significantly decreased compared with control group. But body weight, $1L-1{\beta}$ content in synovial fluids, expression of iNOS and MMP-3 of the treated group were not significantly changed compared with control group. Conclusions: On the basis of these results, we conclude that Gagamsosokmyeong-tang(Jiajianxiaoxuming-tang) has anti-arthritic effects on the MIA induced early stage osteoarthritis in rats.

• Tuberculosis and Respiratory Diseases
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• 제73권1호
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• pp.22-31
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• 2012

Background: Oxidation plays an important role in acute lung injury. This study was conducted in order to elucidate the effect of repetitive post-treatment of N-acetylcysteine (NAC) in lipopolysaccaride (LPS)-induced acute lung injury (ALI) of rats. Methods: Six-week-old male Sprague-Dawley rats were divided into 4 groups. LPS (Escherichia coli 5 mg/kg) was administered intravenously via the tail vein. NAC (20 mg/kg) was injected intraperitoneally 3, 6, and 12 hours after LPS injection. Broncho-alveolar lavage fluid (BALF) and lung tissues were obtained to evaluate the ALI at 24 hours after LPS injection. The concentration of tumor necrosis factor ${\alpha}$ (TNF-${\alpha}$) and interleukin $1{\beta}$ (IL-$1{\beta}$) were measured in BALF. Nuclear factor ${\kappa}B$ (NF-${\kappa}B$), lipid peroxidation (LPO), and myeloperoxidase (MPO) were measured using lung tissues. Micro-computed tomography (micro-CT) images were examined in each group at 72 hours apart from the main experiments in order to observe the delayed effects of NAC. Results: TNF-${\alpha}$ and IL-$1{\beta}$ concentration in BALF were not different between LPS and NAC treatment groups. The concentration of LPO in NAC treatment group was significantly lower than that of LPS group ($5.5{\pm}2.8$ nmol/mL vs. $16.5{\pm}1.6$ nmol/mL) (p=0.001). The activity of MPO in NAC treatment group was significantly lower than that of LPS group ($6.4{\pm}1.8$ unit/g vs. $11.2{\pm}6.3$ unit/g, tissue) (p<0.048). The concentration of NF-${\kappa}B$ in NAC treatment group was significantly lower than that of LPS group ($0.3{\pm}0.1\;ng/{\mu}L$ vs. $0.4{\pm}0.2\;ng/{\mu}L$) (p=0.0001). Micro-CT showed less extent of lung injury in NAC treatment than LPS group. Conclusion: After induction of ALI with lipopolysaccharide, the therapeutic administration of NAC partially attenuated the extent of ALI through the inhibition of NF-${\kappa}B$ activation.