• Title/Summary/Keyword: ELISA system

검색결과 327건 처리시간 0.033초

Lysate of Probiotic Lactobacillus plantarum K8 Modulate the Mucosal Inflammatory System in Dextran Sulfate Sodium-induced Colitic Rats

  • Ahn, Young-Sook;Park, Min Young;Shin, Jae-Ho;Kim, Ji Yeon;Kwon, Oran
    • 한국축산식품학회지
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    • 제34권6호
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    • pp.829-835
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    • 2014
  • Inflammatory bowel disease (IBD) is caused by dysregulation of colon mucosal immunity and mucosal epithelial barrier function. Recent studies have reported that lipoteichoic acid (LTA) from Lactobacillus plantarum K8 reduces excessive production of pro-inflammatory cytokine. In this study, we investigated the preventive effects of lysate of Lb. plantarum K8 in dextran sulfate sodium (DSS)-induced colitis. Male Sprague-Dawley rats were orally pretreated with lysate of Lb. plantarum K8 (low dose or high dose) or live Lb. plantarum K8 prior to the induction of colitis using 4% DSS. Disease progression was monitored by assessment of disease activity index (DAI). Histological changes of colonic tissues were evaluated by hematoxylin and eosin (HE) staining. Tumor necrosis factor-alpha (TNF-${\alpha}$), interleukin-6 (IL-6) levels were measured using enzyme-linked immunosorbent assay (ELISA). The colon mRNA expressions of TNF-${\alpha}$, IL-6, and toll like receptor-2 (TLR-2) were examined by quantitative real-time-transcription polymerase chain reaction (qPCR). Lysate of Lb. plantarum K8 suppressed colon shortening, edema, mucosal damage, and the loss of DSS-induced crypts. The groups that received lysate of Lb. plantarum K8 exhibited significantly decreased levels of the pro-inflammatory cytokines TNF-${\alpha}$ and IL-6 in the colon. Interestingly, colonic expression of toll like receptor-2 mRNA in the high-dose lysate of Lb. plantarum K8 group increased significantly. Our study demonstrates the protective effects of oral lysate of Lb. plantarum K8 administration on DSS-induced colitis via the modulation of pro-inflammatory mediators of the mucosal immune system.

Xylitol Down-Regulates $1{\alpha},25$-Dihydroxy Vitamin D3-induced Osteoclastogenesis via in Part the Inhibition of RANKL Expression in Osteoblasts

  • Ohk, Seung-Ho;Jeong, Hyunjoo;Kim, Jong-Pill;Yoo, Yun-Jung;Seo, Jeong-Taeg;Shin, Dong-Min;Lee, Syng-Ill
    • International Journal of Oral Biology
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    • 제38권3호
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    • pp.127-134
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    • 2013
  • Xylitol is a sugar alcohol with a variety of functions including bactericidal and anticariogenic effects. However, the cellular mechanisms underlying the role of xylitol in bone metabolism are not yet clarified. In our present study, we exploited the physiological role of xylitol on osteoclast differentiation in a co-culture system of osteoblastic and RAW 264.7 cells. Xylitol treatment of these co-cultures reduced the number of tartrate-resistant acid phosphatase (TRAP)-positive multinucleated cells induced by 10 nM $1{\alpha},25(OH)_2D_3$ in a dose-dependent manner. A cell viability test revealed no marked cellular damage by up to 100 mM of xylitol. Exposure of osteoblastic cells to xylitol decreased RANKL, but not OPG, mRNA expression in the presence of $10^{-8}M$ $1{\alpha},25(OH)_2D_3$ in a dose-dependent manner. Furthermore, bone resorption activity, assessed on bone slices in the coculture system, was found to be dramatically decreased with increasing xylitol concentrations. RANKL and OPG proteins were assayed by ELISA and the soluble RANKL (sRANKL) concentration was decreased with an increased xylitol concentration. In contrast, OPG was unaltered by any xylitol concentration in this assay. These results indicate that xylitol inhibits $1{\alpha},25(OH)_2D_3$-induced osteoclastogenesis by reducing the sRANKL/OPG expression ratio in osteoblastic cells.

건동(乾童)이 PTU로 유발된 Rat의 갑상선 기능저하증에 미치는 영향 (The Effects of Zingiberis rhizoma on Hypothyroidism Rat induced by PTU)

  • 강기훈;이병철;안세영;두호경;안영민
    • 대한한방내과학회지
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    • 제27권3호
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    • pp.677-687
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    • 2006
  • Objective : Hypothyroidism is a common disease of the endocrinal system. characterized by fatigue, cold intolerance, bradycardia, and so on. Clinically, Levothyroxine(L-T4) has been usually used for replacement therapy, but it often has side effects. so many hypothyroidism patients wants oriental medical therapy. Zingiberis rhizoma, traditionally has been used in treatment of coldness, fatigue, and bradycardia. In this study. I investigated the therapeutic effects of Zingiberis rhizoma on PTU induced hypothyroidism in rats. Methods : I used two-month-old rats administered PTU and induced with hypothyroidism. After 2 weeks. Zingiberis rhizoma and thyroxine were daily administered, respectively. Body weights was measured every weeks. After 4 weeks, blood samples were taken and analyzed biochemically and T4 and TSH were measured by ELISA kits. Results : In comparison with normal groups, control groups showed hypothyroidism with low T4 and high TSH level. In Zingibris rhizoma administration groups were observed T4 level elevation, this elevation was dependent on the dose of Zingibris rhizoma. Between experimental groups and control groups, there was no difference in TSH level, statistically. Changes of biochemistry were not observed in any experimental groups. Conclusions : These findings suggest that Zingiberis rhizoma makes thyroid cells producing thyroid hormones. There is also a non-toxic effect on the cardiovascular system, liver and kidney function. So, Zingiberis rhizoma should be an effective agents for treating hypothyroidism.

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The increasing hematopoietic effect of the combined treatment of Korean Red Ginseng and Colla corii asini on cyclophosphamide-induced immunosuppression in mice

  • Lee, Yuan Yee;Irfan, Muhammad;Quah, Yixian;Saba, Evelyn;Kim, Sung-Dae;Park, Seung-Chun;Jeong, Myung-Gyun;Kwak, Yi-Seong;Rhee, Man Hee
    • Journal of Ginseng Research
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    • 제45권5호
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    • pp.591-598
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    • 2021
  • Background: Hematopoiesis is the production of blood cells from hematopoietic stem cells (HSCs) that reside in the bone marrow. Cyclophosphamide (CTX) is a chemotherapy drug that suppresses the immune system. Korean Red Ginseng (KRG) and Colla corii asini (CCA) have been traditionally used for boosting the immune system. Methods: HSCs in the bone marrow, and immune cell subtype in splenocytes, PBMCs, and thymocytes were investigated. Serum levels of hematopoietic-related markers were analyzed using ELISA. Protein expression in spleen tissue was analyzed using western blot analysis. Hematoxylin & eosin staining in the femurs of mice were also conducted. Results: The combination of KRG and CCA with a ratio of 3:2 increased HSCs, CD3 and CD8+ T cells in the circulation, and CD3 T cells in the spleen. A ratio of 2:3 (KRG:CCA) increased the thymic regulatory T cells and recovered the CD3 T cells in the spleen and circulation while recovering proteins in the JAK-STAT pathway in the spleen. Overall, blood cell population and differentiating factors vital for cell differentiation were also significantly recovered by all combinations especially in ratios of 3:2 and 2:3. Conclusion: A ratio of 3:2 (KRG:CCA) is the most ideal combination as it recovered the HSC population in the bone marrow of mice.

Mannan-binding lectin of the sea cucumbers Stichopus japonicus has common antigenic determinants with human serum mannan-binding lectin

  • Bulgakov, A.A.;Petrova, I.Yu.;Vakhrusheva, N.M.;Eliseikina, M.G.
    • 한국어업기술학회:학술대회논문집
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    • 한국어업기술학회 2000년도 춘계수산관련학회 공동학술대회발표요지집
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    • pp.530-530
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    • 2000
  • The host defense system or immune system of all modern animals has their roots in very ancient organisms. After analyzing literature data concerning properties of invertebrates and vertebrates lectins we suggest that mechanism of mannans recognition may exist in marine invertebrates, as a universal mechanism for homeostasis maintenance and host defense, and mannan-binding lectins family of vertebrates has ancient precursor, as was shown for another S-type lectins family. We carried out the screening of mannan-binding type lectin among different species of echinoderms inhabiting in Piter the Grate Bay, the sea of Japan. As a result, the C-type lectins (SJL-32) specific for high mannose glycans was isolated from the coelomic plasma of the sea cucumbers Stichopus japonicus by ion-exchange chromatography on a DEAE-Toyopearl 650M, affinity chromatography on a mannan-Sepharose 6B and gel filtration on a Sephacryl S-200. SJL-32 is homodimer with molecular mass about 32 kDa on SDS-PAGE under non-reducing conditions. Protein part of the lectin has high conteins Asn, Glu, Ser. Hemagglutination of trypsin-treated O blood group human erythrocytes by SJL-32 was competitively inhibited by high-branched -D-mannan composed of -1,2 and -1,6 linked D-mannopyranose residues. In contrast, a variety of mono-, oligo-, and polysaccharides composed of residues of galactose and fucose showed absence or little inhibitory activities. The lectin activity strong depends on Ca2+ concentration, temperature and pH. Monospecific polyclonal antibodies were obtained to the lectin. As was shown by ELISA assay, antibodies to SJL-32 cross-reacted with human serum mannan-binding lectin. This data allows making conclusion about common antigenic determinants and structural homology of both lectins. In our opinion, SJL-32 belongs to evolutionary high conservative mannan-binding lectins (MBLs) family and takes part in the host defense against pathogenic microorganisms.

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가미청상보하탕(加味淸上補下湯) 열수 및 에탄올 추출물이 천식 관련 cytokine 분비에 미치는 영향 (The inhibitory effects of hot water and ethanol extract of Gamichungsangbohatang on cytokines related to ashtma)

  • 정승연;이성헌;황준호;이건영;김진주;정희재;이형구;정승기
    • 대한한방내과학회지
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    • 제27권2호
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    • pp.521-532
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    • 2006
  • Backgrounds and Objectives: A major goat in asthma therapy isto reduce or prevent the inflammatory response associated with bronchial hyperresponsiveness, reversible airway obstruction and airway remodelling. Many studies have shown that eosinophilic infiltration is a prominent feathure in the pathophysiology of asthma. The importance of the Presence of eosinophils in the airways of asthmatics has long been recognized, but the mechanism by which these cells are recruited and retained in the lung are only now being elucidated Production of chemotactic cytokines by bronchial epithelial cells may contribute to the local accumulation of eosinophils in Patients with bronchial asthma. This study was designed to investigate the inhibitory effect of extraction of herbal dicoction, Gamichungsangbohatang(GMCSBHT) on eosinophil chemotactic cytokines. Material and Methods:We used water and 70% ethanol extracts of GMCSBHT and pulmonary epithelial cell lines A549(human type II -like epithelial cells). We estimated cytotoxic effects of GMCSBHT, and estimated the effects of water and 70% ethanol extracts of GMCSBHT on chemokines from prestimulated A549 cells by sandwich ELISA. Results: Chemokines of eotaxin, IL-8 were inhibited by GMCSBHT in dose-dependent manner. And ethanol extract of GMCSBHT has more inhibitory effects on eotaxin, IL-8 than hot water extract. Conculusions: These findings indicate that the supression of the expression of chemokines can be accomplished by GMCSBHT treatment, so GMCSBHT may inhibit the iuflammatory process by eosinophils in asthma.

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인간 단핵구 THP-1 세포에서 β-glucan으로 인한 TNF-α 분비 증가 효과 (β-glucan Stimulates Release of TNF-α in Human Monocytic THP-1 Cells)

  • 금보람;현진이;최소희;진지영;정지우;임종민;박동찬;조광근;최은영;최인순
    • 생명과학회지
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    • 제27권11호
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    • pp.1256-1261
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    • 2017
  • ${\beta}$-glucan은 균류의 세포벽, 귀리, 효모, 식물의 구성물질로, 면역 세포의 활성, 전염증성 사이토카인 분비, 항암효능과 같은 면역 체계에 중요한 역할을 한다. 면역계는 건강한 몸 상태의 항상성을 유지한다. 하지만, 병원성 물질이 신체 내로 들어오게 되면 면역 항상성이 무너지게 되고, 질병이 유발될 수 있다. 따라서, 본 연구는 ${\beta}$-glucan이 인간 단핵구 THP-1 세포에서 면역 조절 효과에 이용될 수 있는지를 확인하였다. ${\beta}$-glucan은 THP-1 세포에 다양한 농도를 처리하여 배양하였으며, $TNF-{\alpha}$ mRNA 발현과 단백질 수준을 Real-time PCR와 ELISA을 이용하여 분석하였다. 또한 전사 인자 $NF-{\kappa}B$ p50와 MAPKs 신호 기작 활성을 western blot을 이용하여 분석하였다. ${\beta}$-glucan으로 유도된 MAPKs와 $NF-{\kappa}B$ p50 활성이 증가하였다. ${\beta}$-glucan이 인간 단핵구 THP-1 세포에서 $TNF-{\alpha}$ 생성에 의해 면역 증강 효과를 나타내며, 이는 MAPKs와 $NF-{\kappa}B$ p50 신호 전달을 통해 나타내는 것을 제시한다. 종합적으로, 본 연구는 ${\beta}$-glucan이 인간 단핵구 THP-1 세포를 통해 면역 체계를 향상시킬 것이라고 사료된다.

Polybrominated Diphenyl Ethers Orally Administration to Mice Were Tansferred to Offspring during Gestation and Lactation with Disruptions on the Immune System

  • Hong, Soon-Keun;Sohn, Kyung-Hee;Kim, In-Young;Lee, Jong-Kwon;Ju, Jung-Hun;Kim, Jin-Ho;Lim, Chae-Hyung;Han, Beom-Seok;Jung, Hwa-Chul;Lee, Jin-Yong;Park, Kui-Lea
    • IMMUNE NETWORK
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    • 제10권2호
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    • pp.64-74
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    • 2010
  • Background: The present study was undertaken to examine the immunological effects of pentabrominated diphenyl ether (penta-BDE) and decabrominated diphenyl ether (deca-BDE) on the immune system of the dams and the developmental immune system of the offsprings. Methods: In this study, mated female C57BL/6J mice were orally administered penta-BDE, deca-BDE or corn oil for 5 weeks, from gestational day 6 to lactational day 21. Results: The body weight of PND21 exposed to penta-BDE was significantly decreased relative to control mice, but that of post-natal day 63 (PND63) were recovered. Orally dosed dams with penta-BDE had significantly smaller absolute and relative spleen masses than control mice. Absolute and relative spleen and thymus masses of PND21 exposed to penta-BDE were significantly decreased over control. The exposure of dams and PND21 with penta-BDE reduced the number of splenocytes and thymocytes. As results of hematologic analysis, percentage WBC and percentage neutrophils increased in dams with deca-BDE. Splenic T cell proliferation in dams and PND21 exposed to penta-BDE was increased, and there were no significant difference in splenic B cell proliferation in all treatment groups. As results of flow cytometric analysis of splenocyte, percentage total T cell, Th cell and Tc cell in PND21 exposed to penta-BDE was slightly increased, and percentage macrophage in dams and PND21 exposed to deca-BDE was decreased. The ELISA results of antibody production show no significant difference in all treatment groups relative to controls. Conclusion: These results imply that PBDEs given to the dam were transferred to the offspring during gestation and lactation, and PBDEs transferred from the dam affect immune system of offspring.

해열탕(解熱湯)이 LPS로 자극된 대식세포에 있어 염증관련 Cytokine 발현억제에 미치는 효과 (The Inhibitory Effects of Haeyeol-tang in Expression of Pro-inflammatory Cytokine on LPS-stimulated THP-1 Cells)

  • 이병삼;김홍렬;김진주;정승기;이형구;정희재
    • 대한한방내과학회지
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    • 제29권2호
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    • pp.334-347
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    • 2008
  • Background and Objective : Haeyeol-tang, composed of Houttuyniae Herba, Lonicerae Flos, Taraxaci Herba, and Scrophulariae Radix, is widely used for alleviating the symptom of various kinds of inflammatory pulmonary disease, including asthma and COPD. We want to know whether Haeyeol-tang has an anti-inflammatory effect by analyzing expression of pro-inflammatory cytokines. Materials and Methods : We differentiated the THP-1 cells into macrophage-like cells by treatment with PMA. Inflammation was induced by treatment with LPS and PMA. We found the safe concentration of Haeyeol-tang by using MTS assay and used PD98059 as a negative control for comparison of anti-inflammatory effect of Haeyeol-tang. Results : The RT-PCR analysis results show that the cell survival rate is over 100% within 1 ng/mL to 1 ug/mL of Haeyeol-tang and begins to decrease under 100% at 10 ug/mL. The gene expression of $IL-1{\beta}$, IL-6, IL-8, IL-10, $TNF-{\alpha}$ and $TGF-{\beta}$ levels were down-regulated when Haeyeol-tang was treated at concentrations between 1 ng/mL an 1 ug/mL on monocyte-derived macrophages. Interestingly, 1 ug/mL Haeyeol-tang-treated samples showed that the transcriptional activities of IL-8, $TNF-{\alpha}$, IL-10 and $TGF-{\beta}$ were more down-regulated than those of PD098059 $(TNF-{\alpha}$ inhibitor). At protein level, the ELISA analysis results showed that there were more remarkable (p<0.001) decreases in expression of $IL-1{\beta}$, IL-6, IL-8 and $TNF-{\alpha}$ on both the 1 ug/mL Haeyeol-tang-treated group and the PD98059-treated group than the LPS-treated group. Conclusion : We conclude that Haeyeol-tang has an anti-inflammatory effect by inhibiting expression of pro-inflammatory cytokines and chemokines at mRNA and protein levels. These results may provide us a promising way to care for general inflammatory diseases as well as inflammatory pulmonary disease, including asthma and COPD, with further clinical study.

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가미청상보하탕(加味淸上補下湯)의 물과 에탄올 추출법에 따른 항(抗)알레르기 효과(效果)에 대(對)한 연구(硏究) (Studying of the Effects of Gamichungsangboha-tang water-extract and ethanol-extract on $IFN{\gamma}$, IL-4, T-bet, GATA-3, c-maf in Asthma Model)

  • 허태석;고은정;정희재;이형구;배현수;정승기
    • 대한한방내과학회지
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    • 제27권1호
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    • pp.40-54
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    • 2006
  • Objective: This study aimed to identify the different effects of GMCSBHT water-extract and ethanol-extract on Th1/Th2 differentiation by monitoring Th1/Th2 specific cytokine secretion patterns and the transcriptional activities of T-bet, GATA-3, c-maf, $INF{\gamma}$ and IL-4. Materials and Methods: Spleen cells from eight week-old BALB/c mice were cultured in GMCSBHT extracts containing medium without activation for 24 hours and with activation for 48 hours. CD4+ T cells were isolated and mRNA expression levels of $INF{\gamma}$, IL-4, T-bet, GATA-3, c-maf by RT-PCR and secretion cytokines levels of $IFN{\gamma}$, IL-4 by ELISA were analyzed. Results: GMCSBHT extracts didn't have mitogenic effects on the unstimulated CD4+ T cells. In Th1 skewed condition, GMCSBAHT water extract had no significant effects on mRNA expression levels of $IFN{\gamma}$, T-bet and c-maf, but inhibited mRNA expression levels of IL-4, GATA-3. It showed significantly increased secretion cytokine levels of $IFN{\gamma}$, but had no significant effect on secretion cytokine levels of IL-4. In Th2 skewed condition, GMCSBHT ethanol extract inhibited mRNA expression levels of $INF{\gamma}$, IL-4, GATA-3 and c-maf significantly, but had no significant effects on mRNA expression levels of T-bet. It had no significant effects on secretion cytokine levels of $INF{\gamma}$, but showed remarkable inhibitory effects on secretion cytokine levels of IL-4. Conclusion: Results suggest that on Th1/Th2 deviation, GMCSBHT water extract has both amplifying effects on Th1 differentiation and inhibitory effects on Th2, but GMCSBHT ethanol extract has stronger inhibitory effects on Th2 differentiation than on Th1.

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