• 제목/요약/키워드: ELISA

검색결과 2,969건 처리시간 0.034초

교맥의 RBL-2H3 비만세포 탈과립과 cytokine 생산 억제 효과 (Inhibitory effect of Fagopyrum esculentum on degranulation and production of cytokine in RBL-2H3 cells)

  • 강경화;이승연
    • 한방안이비인후피부과학회지
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    • 제25권3호
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    • pp.1-12
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    • 2012
  • Objectives : Fagopyrum esculentum(FE) has been used for removal of inflammation of internal organs and treatment of sore and ulcer by heat toxin in Korean herbal medicines. In this study, To investigated the protective effect of FE on allergic response, we determined whether FE inhibits allergic response. Methods : The effect of FE was analyzed by ELISA, RT-PCR and Western blot in RBL-2H3 cells. We investigated cell viability, ${\beta}$-hexosaminidase, as a marker of degranulation, cytokne, and intracellular ROS and MAPK and NF-${\kappa}B$ signaling. Results : We found that FE suppressed ${\beta}$-hexosaminidase release, the production of IL-4 and TNF-${\alpha}$ and intracellular ROS level in RBL-2H3 by the anti-DNP IgE plus DNP-HSA stimulation. FE also significantly inhibited cytokine mRNA expressions, such as IL-$1{\beta}$, IL-2, IL-3, IL-4, IL-5, IL-6, IL-13, TNF-${\alpha}$ and GM-CSF in RBL-2H3. In addition, PF suppressed the phospholyation of ERK1/2, JNK1/2, p38 and $I{\kappa}B{\alpha}$ and NF-${\kappa}B$ signal transduction pathway. Conclusions : Our results indicate that FE protects against allergic response and exerts an anti-inflammatory effect through the inhibition of degranulation and production of cytokines and ROS via the suppression MAPK and NF-${\kappa}B$ of signal transduction. Abbrevations : FE, Fagopyrum esculentum; RBL-2H3, rat basophilic leukemia cell line; ROS, reactive oxygen species; MAPK, Mitogen-activated protein kinase; $NF{\kappa}B$, nuclear factor ${\kappa}B$; $TNF{\alpha}$, Tumor necrosis factor alpha; GM-CSF, Granulocyte macrophage colony-stimulating factor; ERK, extracellular-signal-regulated kinase; JNK, c-Jun NH2-terminal kinase; p38, p38 MAP kinase; $I{\kappa}B{\alpha}$, inhibitory-kappa B alpha.

폴리페놀 함량이 높은 Hylotelephium erythrosticum 추출물의 항산화 및 항염 활성 (Antioxidant and anti-inflammatory effects of phenolic rich Hylotelephium erythrostictum extracts)

  • 김상숙;박경진;이승은;이정훈;최영훈
    • 한국식품저장유통학회지
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    • 제24권6호
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    • pp.842-848
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    • 2017
  • 피부병 치료에 사용되던 꿩의비름 추출물을 항산화 및 항염 효능을 지니는 천연물 소재로 활용하기 위해 꿩의비름의 뿌리(HER), 잎(HEL), 줄기(HEB), 지상부(HEAG) 추출물에 대해 항산화 활성을 측정한 결과 지상부 추출물인 HEAG에서 DPPH, ABTS, superoxide anion radical 소거 활성이 가장 우수하였으며, FRAP 활성 역시 HEAG에서 가장 우수하였다. 항산화 활성이 가장 우수한 HEAG의 총 폴리페놀 함량을 측정한 결과 $235.5{\pm}18.96mg/g$ gallic acid equivalent로 항산화 및 항염 활성을 갖는 것으로 알려진 페놀성 화합물을 다량 함유하고 있는 것을 확인할 수 있었다. 또한 마우스 대식세포인 RAW264.7 세포를 이용하여 항염 활성에서는 NO의 생성을 억제하였으며 염증성 사이토카인인 $IL-1{\beta}$의 생성 억제 화성이 가장 높았다. 꿩의비름 추출물은 항산화 및 항염 활성의 천연물 소재로 활용이 가능할 것으로 여겨진다.

홍삼의 생체 내 사이토카인 분비에 대한 면역조절효과 (Immunomodulating Effects of Red Ginseng on the Regulation of Cytokine Release in vivo)

  • 손은화;윤재웅;구현정;박대원;정용준;남궁승;한효상;강세찬
    • 한국자원식물학회지
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    • 제25권5호
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    • pp.578-585
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    • 2012
  • 홍삼의 6주간의 복강투여가 생체 내 면역계에 미치는 영향을 알아보기 위하여 면역계를 구성하는 주요 장기인 비장과 흉선의 무게변화와 성숙된 B세포와 T세포가 많이 분포되어 있는 비장세포를 배양하여 mitogen에 대한 비장세포의 세포증식능력을 연구하였다. 또한, 생체내 투여되는 홍삼이 면역반응의 매개역할을 하는 분비성 사이토카인의 조절 양상을 확인하기 위하여 생쥐의 혈청에서 분리한 사이토카인의 생성변화를 측정하였다. 연구결과 홍삼의 투여는 비장과 흉선의 무게를 증가시켰고, 비장세포내의 B세포와 T세포의 증식능력에도 유의적인 효과를 나타내었다. 혈청 내 분비된 T세포, B세포 및 대식세포가 분비하는 사이토카인의 농도변화에서도 홍삼 투여군은 면역계를 활성화시키는 IFN-${\gamma}$, TNF-${\alpha}$, IL-2, IL-6 및 IL-12의 분비량을 모두 증가시켰으며, 면역억제성 사이토카인으로 알려진 IL-10의 분비변화에는 영향을 미치지 않았다. 이와 같은 결과는 홍삼의 투여가 면역 반응계 전반을 조절하는 사이토카인의 생성에 영향을 미치며 모두 면역계를 활성화시키는 방향으로 작용시키는 것을 의미한다. 특히, 흉선과 비장의 무게지수가 증가되었다는 것은 세포증식 등의 변화에 영향을 미침으로써 면역계를 활성화시키는 것으로 생각되며, 특히 흉선의 무게 증가와 ConA에 대한 T세포 증식능력의 유의성 변화 및 T세포가 분비하는 IL-2, IFN-${\gamma}$ 등의 사이토카인의 증가는 홍삼의 생체 내 투여가 T세포의 활성화에 크게 영향을 미치는 것으로 사료된다.

Adipose-Derived Stem Cells Suppress Inflammation Induced by IL-1β through Down-Regulation of P2X7R Mediated by miR-373 in Chondrocytes of Osteoarthritis

  • Jin, Rilong;Shen, Miaoda;Yu, Liedao;Wang, Xuanwei;Lin, Xiangjin
    • Molecules and Cells
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    • 제40권3호
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    • pp.222-229
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    • 2017
  • Adipose-derived stem cells (ADSCs) were previously considered to have an anti-inflammatory effect, and Interleukin-$1{\beta}$ ($IL-1{\beta}$) was found to be a pro-inflammatory factor in chondrocytes, but the mechanism underlying ADSCs and $IL-1{\beta}$ is unclear. In this study, we investigate whether P2X7 receptor (P2X7R) signalling, regulated by microRNA 373 (miR-373), was involved in the ADSCs and $IL-1{\beta}$ mediated inflammation in osteoarthritis (OA). Chondrocytes were collected from 20 OA patients and 20 control participants, and ADSCs were collected from patients who had undergone abdominal surgery. The typical surface molecules of ASDCs were detected by flow cytometry. The level of nitric oxide (NO) was determined by Griess reagent. Concentrations of prostaglandin E2 (PGE2), interleukin 6 (IL-6), matrix metallopeptidase 3 (MMP-3) were detected by enzyme-linked immunosorbent assay (ELISA). The expressions of IL-6, MMP-3, miR-373 and P2X7R were determined by real-time polymerase chain reaction (PCR), and Western blot was used to detect the protein expression of P2X7R. The typical potential characters of ADSCs were verified. In chondrocytes or OA tissues, the miR-373 expression level was decreased, but the P2X7R expression was increased. $IL-1{\beta}$ stimulation increased the level of inflammatory factors in OA chondrocytes, and ADSCs co-cultured with $IL-1{\beta}$-stimulated chondrocytes decreased the inflammation. OA chondrocytes transfected with the miR-373 inhibitor increased the inflammation level. The miR-373 mimic suppressed the inflammation by targeting P2X7R and regulated its expression, while its effect was reversed by overexpression of P2X7R. $IL-1{\beta}$ induced inflammation in OA chondrocytes, while ADSCs seemed to inhibit the expression of P2X7R that was regulated by miR-373 and involved in the anti-inflammatory process in OA.

삼기음(三氣飮)이 흰쥐의 Monosodium Iodoacetate 유발 골관절염에 미치는 영향 (Effects of Samgi-eum(Sānqì-yǐn) Treatment on the Monosodium Iodoacetate-induced Osteoarthritis in Rats)

  • 이경무;정수현;김순중;서일복
    • 한방재활의학과학회지
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    • 제18권2호
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    • pp.17-31
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    • 2008
  • Objectives : This study was carried out to investigate the effects of Samgi-eum($S{\bar{a}}nq{\grave{i}}-y{\check{i}}n$) on the monosodium iodoacetate-induced osteoarthritis in rats. Methods : Osteoarthritis was induced by injection of monosodium iodoacetate intraarticularly in both knee joints. Arthritic rats were divided into control and treated group. Control group were taken distilled water for 20days. Treated group were taken extracts of Samgi-eum($S{\bar{a}}nq{\grave{i}}-y{\check{i}}n$) by orally for the same duration. Normal group were injected normal saline and taken distilled water. Body weights were measured at 0, 5th, 10th, 15th, 20th day after injection. At the end of the experiment, gross and histopathological examination on the articular cartilages of the knee joints were performed. Proteoglycan contents of articular cartilages were analyzed by safranine O staining method. The contents of $TNF-{\alpha}$, $IL-1{\beta}$ and IL-6 in synovial fluids were analyzed by ELISA method. Results : 1. Body weights of the treated group were significantly increased compared with control at 20days after injection. 2. Grossly, the severity of osteoarthritis in the treated group were alleviated compared with control. 3. Histopathologically, degenerative and necrotic lesion of articular cartilages in the treated group were alleviated compared with those of the control and histopathological scores of treated group were significantly decreased compared with control. 4. PG contents in articular cartilages of the treated group were significantly increased compared with control. 5. $TNF-{\alpha}$ contents in synovial fluids of the treated group were significantly decreased compared with control. Conclusions : According to above results, Samgi-eum($S{\bar{a}}nq{\grave{i}}-y{\check{i}}n$) has anti-arthritic effects on the monosodium iodoacetate-induced osteoarthritis in rats. And it is related with reduced secretion of $TNF-{\alpha}$ from osteoarthritic chondrocytes and synovial membranes.

Effect of Lipopolysaccharide (LPS) on Mouse Model of Steroid-Induced Avascular Necrosis in the Femoral Head (ANFH)

  • Ryoo, Soyoon;Lee, Sukha;Jo, Seunghyun;Lee, Siyoung;Kwak, Areum;Kim, Eunsom;Lee, Jongho;Hong, Jaewoo;Jhun, Hyunjhung;Lee, Youngmin;Sobti, Anshul Shyam;Kim, Soohyun;Oh, Kwang-Jun
    • Journal of Microbiology and Biotechnology
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    • 제24권3호
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    • pp.394-400
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    • 2014
  • Avascular necrosis of the femoral head (ANFH) is commonly observed in patients treated with excessive glucocorticoid (GC). Single administration of lipopolysaccharide (LPS) has shown to induce immune stimulatory factors. However, the effect of repeated administration of LPS on GC-induced ANFH has not been studied. Thus, the purpose of this study was (i) to examine the cytokine profile induced by repeated LPS administrations and (ii) to test the effect of repeated LPS treatments on GC-induced ANFH. A mouse necrosis model of ANFH was designed by chronic GC administration with co-treatment of LPS. Mice body weights in the LPS/prednisolone (PDN) co-treated group were lower than that of the untreated control group, but spleen weights were greater than the control group. The levels of IL-6, $TNF{\alpha}$, and IL-33 in the liver and spleen of the LPS/PDN group were lower than the untreated control group, whereas $TNF{\alpha}$ level in the femoral head of the LPS/PDN group increased. Collectively, the effect of repeated LPS on the pathogenesis of GC-induced ANFH was associated with the $TNF{\alpha}$ level in the femoral head, but the pathogenesis did not correspond to cytokine levels in immune tissues.

NO와 Pro-Inflammatory Cytokine의 억제를 통한 호라복(胡蘿蔔)의 항염증효과 (Roots of Daucus carota sativa abrogates acute phase of Inflammation by the Inhibition of NO and Pro-Inflammatory Cytokine Production)

  • 이동진;박상미;황보민;정태영;김상찬;지선영
    • 한방안이비인후피부과학회지
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    • 제26권2호
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    • pp.45-57
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    • 2013
  • Objectives : Daucus carota sativa has been frequently used as food supplements in many of the Asian countries, and a nutritional medical drug in traditional medicine. This research investigated the effects of Daucus carota sativa extract (DCE) on acute phases of inflammation in Raw 264.7 cells treated with lipopolysaccharide (LPS) in terms of the inhibition of nitric oxide (NO), prostaglandin $E_2$ ($PGE_2$) and pro-inflammatory cytokines production. Methods : NO, $PGE_2$, tumor necrosis factor (TNF)-${\alpha}$, interleukin-$1{\beta}$ and interleukin-6 contents were assayed by ELISA, and expressions of inflammation-related proteins such as inducible NO synthase (iNOS) were determined by immunoblot analyses. Results : DCE treatment attenuated the LPS ability to increase the productions of NO and $PGE_2$ as well as the protein level of iNOS in a concentration-dependent manner. Consistently, treatment of the cells with DCE suppressed the production of TNF-${\alpha}$, interleukin-$1{\beta}$ and interleukin-6. DCE also caused decreases of inhibitor of ${\kappa}B{\alpha}$ phosphorylation induced by LPS in the cells, which means DCE inhibition of NF-${\kappa}B$ activity. Furthermore, DCE blocked LPS-induced phosphorylation of p38 and SAPK/JNK. Conclusion : This study showing here may be of help to understand the action mechanism of DCE, and provide the information for the medical use of Daucus carota sativa for the inflammatory disease.

개 심장사상충을 진단하기 위한 중합연쇄반응검사 (PCR)의 진단적 특성 평가 (Evaluation of Diagnostic Performance of a Polymerase Chain Reaction for Detection of Canine Dirofilaria immitis)

  • 박선일;김두
    • 한국임상수의학회지
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    • 제24권2호
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    • pp.77-81
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    • 2007
  • 본 연구는 개에서 심장사상충을 검출하기 위하여 표준검사를 적용하지 않은 상황에서 중합연쇄반응검사 (PCR)의 진단 능력을 평가하였다. 효소면역검사법 (ELISA)과 PCR 검사를 동시에 사용한 경우 PCR 검사의 민감도와 특이도는 두 검사의 조건부 독립을 가정한 상태에서expectation-maximization (EM) 알고리즘을 이용한 최대우도법과 Bayesian 기법으로 두 집단 검사 모형으로 분석하였다 2002-2004년 기간 중 심장사상충검사 결과를 기록한 의무기록에서 무작위로 266개 결과를 추출하여 133개씩 2회의 시험으로 배치하였다. 2회의 분석결과를 종합할 때 EM 알고리즘에서 PCR 검사의 민감도와 특이도는 각각 96.4-96.7%와 97.6-98.8%, Bayesian기법에서는 94.4-94.8h와 97.1-98%로 추정되었다. PCR 검사는 심장사상충을 스크리닝하는 도구로 유용하며, 표준검사를 적용하지 않은 상황에서 진단검사의 특성을 추론하는 방법으로 Bayesian 기법은 매우 유용함을 확인하였다.

마우스에서 살모넬라 감염증 예방을 위한 신개념 Salmonella Typhimurium 불활화 사균체에 최적 adjuvant 선택을 위한 효능 비교 시험 (Comparative evaluation to select optimal adjuvant of novel type Salmonella Typhimurium inactivated bacteria for protecting Salmonella infections in a murine model)

  • 문자영;;김원경;이준우;조영규;곽길한;박병용;허진
    • 한국동물위생학회지
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    • 제43권2호
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    • pp.89-97
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    • 2020
  • This study was carried out to examine a novel inactivated Salmonella Typhimurium (S. Typhimurium) vaccine candidate for protection of mice against salmonellosis by immunization of BALB/c mice using various type adjuvant. The novel type-inactivated vaccine candidate was constructed by adding Chlorhexidine digluconate solution. BALB/c mice were divided into 6 groups of 15 mice apiece. The mice were intramuscularly (IM) primed at 6 weeks of age and were IM boosted 8 weeks of age. Groups A and B mice were injected with sterile phosphate-buffered saline as controls; group C mice were inoculated with 5×108 cells/100 µL of formalin-inactivated S. Typhimurium cells and adjuvant ISA70; groups D~F mice were immunized with 5×108 cells/100 µL of the inactivated vaccine candidate and adjuvant ISA70, adjuvant IMS1313 and adjuvant IMS1313 containing 30 ㎍/mL of GI24, respectively. All mice (except group A mice) were orally challenged with a virulent S. Typhimurium strain at 10 weeks of age. Mice from groups C-F had significantly increased IgG levels compared to control groups (A-B) mice. The levels of splenocyte IFN-γ and IL-4 in mice of all groups were measured by ELISA, resulting in increased immunity in group F mice compared to those of groups A-E mice. These data suggested that systemic and cell-mediated immune responses were highly induced by IM immunization with the vaccine candidate and adjuvant IMS1313 containing GI24. Furthermore, clinical signs such as death were observed in only 20% of group F mice after virulent Salmonella strain challenge, however, groups B and C (100%), and groups D and E (60%) mice died. This data suggested that mice immunized by intramuscular prime and booster with this vaccine candidate and adjuvant IMS1313 containing GI24 effectively protected mice from salmonellosis.

보중익기탕 가미방(補中益氣湯 加味方)과 선방패독탕(仙方敗毒湯)이 기도 점액의 분비와 뮤신 유전자발현에 미치는 영향 (Effects of Bojung-ikgitang-gamibang and Seonbang-paedoktang on Secretion of Airway Mucus and Expression of Mucin Gene)

  • 정창호;한재경;김윤희
    • 대한한방소아과학회지
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    • 제21권3호
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    • pp.33-55
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    • 2007
  • Objectives In the present study, the author intended to investigate whether bojung-ikgitang-gamibang(BJGB) and seonbang-paedoktang(SBPT) significantly affect in vivo and in vitro mucin secretion from airway epithelial cells. Methods In vivo experiment, mice's mucin which is on a hypersecretion of airway mucin, mice's tracheal goblet cells in hyperplasia and mice's intraepithelial mucosubstances were exposed with SO2for3weeks. Effects of orally-administered BJGB and SBPT during 1 week on vivo mucin secretion and hyperplasia of tracheal goblet cells were assessed by using both enzyme-linked immunosorbent assay(ELISA) and staining goblet cells with alcian blue. In vitro experiment, confluent hamster tracheal surface epithelial(HTSE) cells were metabolically radiolabeled with 3H-glucosamine for 24hrs and chased for 30 min in the presence of each agent to figure out the effectiveness of 3H-mucin secretion. Total elution profiles of control spent media and treatment sample through Sepharose CL-4B column were analyzed. The effects of each agent on contractility of isolated tracheal smooth muscle and effects of each agent on MUC5AC gene expression in cultured HTSE cells were investigated. Also, possible cytotoxicities of each agent were assessed by measuring lactate dehydrogenase(LDH) release. Additionally, effects of BJGB and SBPT on both MUC5AC gene expression in cultured HTSE cells and TNF- or EGF-induced MUC5AC gene expression in human airway epithelial cells (NCI-H292) were investigated. Results (1) BJGB and SBPT inhibited hypersecretion of in vivo mucin. SBPT also inhibited the increase the number of goblet cells. However, BJGB did not affect the increase of number of goblet cells; (2) BJGB significantly increased mucin secretion from cultured HTSE cells, without significant cytotoxicity, and chiefly affected the 'mucin' secretion; (3) SBPT did not affect mucin secretion from cultured HTSE cells without significant cytotoxicity, and also did not affect the secretion of the other releseable glycoproteins; (4) BJGB and SBPT did not affect Ach-induced contraction of isolated tracheal smooth muscle; (5) SBPT significantly inhibit the expression levels of MUC5AC gene and BJGB significantly increased the expression levels of MUC5AC gene in both HTSE cells and NCI-H292 cells. Conclusions BJGB and SBPT can not only affect the secretion of mucin but also affect the expression of mucin gene. The author suggests that the effects BJGB and SBPT with their components should be further investigated and it is highly desirable to find from oriental medical prescriptions, novel agents which might regulate hypersecretion of mucin from airway epithelial cells.

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