• 제목/요약/키워드: cELISA

검색결과 678건 처리시간 0.028초

조절 T세포에 미치는 치자(梔子)의 효과 (Research on the Effect of Gardeniae Fructus on Regulatory T Cell Stimulation)

  • 서산;정희재;정승기
    • 대한한방내과학회지
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    • 제31권2호
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    • pp.189-200
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    • 2010
  • Objectives : Regulatory T cells can reduce inflammation and allergic reactions through their inhibitory functions. Gardeniae Fructus(GF) is a Heat-clearing herb used in traditional Korean medicine, and a wide range of studies on its antiinflammatory effects are being carried out. The authors investigated the effect that Gardeniae Fructus has on regulatory T cells. Methods : The authors screened 14 herbs for their effects on regulatory T cells. 100mg of each herb were separately dissolved in 1ml of sterile saline and the supernatant was harvested after 10 minutes of centrifuge at 15,000 rpm. The supernatant was filtered through a 0.2 ${\mu}m$ syringe filter, and the resulting stock was refrigerated at $4^{\circ}C$. The stock was diluted before testing and used at a final concentration of $0.01{\mu}g/ml$. CD4+CD25+ T cells from healthy BALB/c spleens were used as natural regulatory T cells (nTreg), and CD4+CD25- T cells were used as reactive T cells. CD4+CD25+ and CD4+CD25- T cells were activated with anti-CD3e ($10{\mu}g/m{\ell}$)/anti-CD28 ($1{\mu}g/m{\ell}$) and cultured. IL-10 from supernatant of the culture medium was measured by IL-10 cytokine ELISA. The percentages, cell numbers, phenotype and function of CD4+CD25+ Treg cells were determined by flow cytometry. Results : Gardeniae Fructus was shown to be the most potent herb among the 14 herbs tested for suppressing CD4+CD25- reactive T cell proliferation by stimulating CD4+CD25+ natural regulatory T cells. Gardeniae Fructus induces IL-10 secretion increase by stimulating CD4+CD25+ natural regulatory T cells, and indirectly suppresses CD4+CD25- reactive T cell proliferation through increasing CD25 (IL-2 receptor $\alpha$) expression and thus promoting bonding with IL-2. Gardeniae Fructus did not directly affect CD4+CD25- reactive T cell proliferation. Conclusions : Gardeniae Fructus suppressed reactive T cell proliferation through inducing increases in IL-10 secretion and CD25 (IL-2 receptor $\alpha$) expression.

마행감석양가감방(麻杏甘石揚加減方)이 천식모델생쥐의 면역세포 및 사이토카인에 미치는 영향 (Effects of Mahaenggamseok-tang-gagambang on Immune Cells and Cytokines in OVA-Induced Asthmatic Mice)

  • 박길병;박양춘
    • 동의생리병리학회지
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    • 제23권3호
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    • pp.590-598
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    • 2009
  • The purpose of this research is to evaluate the effect of Mahaenggamseok-tang-gagambang (MGTG) on airway hyper- responsiveness (AHR), immune cells, cytokines and lung tissue in OVA-induced asthmatic mice. C578L/6 mice were injected, inhaled and sprayed with OVA for 12 weeks (3times a week) for asthma sensitization and challenge. Two experimental groups were treated with different concentrations of MGTG (400 mg/kg and 200 mg/kg) extract and cyclosporin A (10 mg/kg) for the later 8 weeks. Enhanced pause (Penh) levels were measured by whole body plethysmography. Immune cells were analyzed by flow cytometer in peripheral blood monocyte cell (PBMC) and lung cells. The IL-1b, IL-12, IFN-${\gamma}$, OVA-lgE, IL-4, IL-5, TNF-${\alpha}$ were analyzed by ELISA kit in serum and splenocyte+a-cCDS/a-CD28. Enhanced pause (Penh) levels of the MGTG groups (400 mg/kg and 200 mg/kg) were decreased significantly compared with that of control group. The numbers of MGTG groups (400 mg/kg and 200 mg/kg) on lung total cells were decreased significantly compared with that of control group. The numbers of MGTG groups (400 mg/kg and 200 mg/kg) on $CD3^+/CD69^+$, $B220^+/CD22^+$, $B220^+/CD23^+$, $B220^+/lgE^+$, $CCR3^+$ cells were decreased significantly compared with that of control group. The number of MGTG group (400 mg/kg) on $CD3^+/CD49b^+$ cells was decreased significantly compared with that of control group. The level of MGTG groups (400 mg/kg and 200 mg/kg) on IL-4, IL-5, IL-12, TNF-${\alpha}$, OVA-lgE were decreased significantly compared with that of control group. The level of MGTG group (400 mg/kg) on IL-1b, IL-1S, OVA-lgE were decreased significantly compared with that of control group. These results demonstrate that MGTG could be a desirable alternative therapy for allergic asthma by inhibiting the expression of immune cells, the activation of inflammatory mediator.

젖소의 난소질환별 Milk Progesterone 수준에 관한 연구 (A study on the concentrations of milk progesterone in dairy cattle with ovarian reproductive disorders)

  • 조정연;이병한;강영선;김진영;육순학;최석화;김종배;윤화중;정병현
    • 대한수의학회지
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    • 제35권1호
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    • pp.159-167
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    • 1995
  • The relationship between palpable ovarian structure and milk progesterone levels were determined in 144 dairy cows. Depending on the ovarian structure and diseases were divided into two groups, Group I (absence of functional luteal tissue in ovary and <2ng/ml in milk progesterone levels) and Group II(presence of functional luteal tissue in ovary and ${\geq}2ng/ml$ in milk progesterone levels) 1. Among 69 cows of group I, dysfunction of ovary, atropy of ovary, follicle is ovary, follicular cyst and corpus luteum albicans were 17(11.8%), 19(13.2%), 14(9.7%), 3(2.1%) and 16 cows(11.1%), and among 75 cows of group II, corpus luteum A, B and C were 16(11.1%), 17(11.8%) and 42 cows(29.2%), respectively. 2. In Group I, milk progesterone concentrations were <1ng/ml in 55 cows(79.9%). Conversely in Ggroup II, milk progesterone concentrations were ${\geq}4ng/ml$ in 55 cows(73.3%). 3. The mean(${\pm}SE$) concentrations of milk progestsrone in the Group I and II were $1.62{\pm}0.45$ and $7.64{\pm}0.68ng/ml$, respectively, and CR test showed the difference in milk progesterone concentrations between the two groups to be statistically significant(p<0.01). 4. The mean(${\pm}SE$) concentration of milk progesterone in cows with corpus luteum A, B and C were $8.11{\pm}1.83$, $8.48{\pm}1.30$ and $7.12{\pm}0.82ng/ml$, respectively, there was no significant relationship between palpable corpora luteum structure and milk progesterone concentration. 5. The accuracy of ovarian diagnosis was 82.6 and 20.2% in the Group I and II, respectively, and Chi-square test showed the difference in accuracy between the two groups to be statistically significant (p<0.001). 6. The agreement between the rectal palpation and milk progesterone concentrations in ovarian disease was 50%.

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지이초(地耳草) 추출물이 OVA로 천식이 유발된 생쥐의 폐세포에 미치는 영향 (Effects of Extract of Hyperici Japonici Herba on Lung Cells in Asthma-indused Mice by OVA Exposure)

  • 이영용;서영배;이영철;서부일;노성수
    • 대한본초학회지
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    • 제23권1호
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    • pp.75-83
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    • 2008
  • Objectives : The present study was carried out to investigate the effect of Hyperici Japonici Herba on the proliferation and activation of eosinophils which were prepared from lung cells of asthma-induced mice by ovalbumin(OVA) treatment. Methods : C57BL/6 mouse was exposed to OVA three times a week for 6 weeks. The mouse lung tissues were dissected out, chopped and dessiciated with collagenase(1${\mu}g$/ml). Eosinophils were activated by rIL-3/rmIL-5 co-treatments. The lung cells were treated with extract of Hyperici Japonici Herba(EHH), incubated for 48 hr at $37^{\circ}C$, and analyzed by flow cytometer. ELBA, RT-PCR, immunocytochemistry stain. Results : The cell number ratio of granulocyte, $CD3e^-$/$CCR3^+$, $CD3e^+$/$CD69^+$, $CD4^+$, $CD23^+$/$B220^+$ cells was increased in rmIL-5/rIL-3 treated control group compared to the normal group. Cells numbers in the experimental animal group treated with EHH was all decreased. In ELISA analysis, IL-4, IL-5, IL-13 protein levels and histamine release level were greatly increased in the control group compared to the normal animal group, then significantly decreased in the experimental group with 100 ${\mu}g$/ml of EHH treatment. In RT-PCR analysis, the HT value of IL-4, IL-5, IL-13, CCR3, Eotaxin were increased in the control group compared to the normal animal group, then decreased in the experimental group with 100 ${\mu}g$/ml of EHH treatment. And eosinophil proliferation levels were 18847${\pm}$1527(cpm) in the control group, 4676${\pm}$972(cpm) in the positive control group, and 8675${\pm}$159(cpm), 11352${\pm}$1005(cpm), 14325${\pm}$677(cpm) in the experimental group with 100 ${\mu}g$/ml, 10 ${\mu}g$/ml, 1 ${\mu}g$/ml of EHH treatment. Conclusions : The present data suggested that Hyperici Japonici Herba may have an effects on the inhibition of parameters associated with asthma responses in eosinpophils, and thus implicate the possibility for the clinical application of EHH.

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부동화 스트레스 유도 마우스 모델에서 도두(刀豆), 우방근(牛蒡根) 복합물의 면역증진 작용 (Enhancement of Immune Activities of Canavalia gladiata & Arctium lappa complexes in immobilization stress mouse model.)

  • 이지은;노성수;김한영;김근회;김승형
    • 대한본초학회지
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    • 제32권1호
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    • pp.1-13
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    • 2017
  • Objective : Soybeans of Canavalia gladiata(CG) and root of Arctium lappa(AL) have been reported to have anti-inflammatory, antioxidant effect. However, the immunoregulatory mechanisms of its combinational prescription remain a matter of considerable debate. In the current study, we investigated whether CG and AL and its combinational prescription(CG+AL) regulate immune system using chronic immobilization-stress mouse model. Methods : C57BL/6J mice fixed for 2 hours into immobilization tube after CG, AL, CG+AL oral administration after 2 hours daily for 21 days. After every experiment has ended the C57BL/6J mice were sacrificed on 22 days. The production of Serotonin and Cortisol, lgA were observed by ELISA method, The proportion of immune cells such as T/B cell and macrophage, NK cell were measured by FACS. Then, Real-time PCR was performed to measure the mRNA expression of Inflammatory cytokines(IL-1beta, IL-6, TNF-a) and T cell activation cytokines(IL-2, IL-10, IFN-gamma, IL-12p35 / p40). Result : When chronic immobilization-stress mouse model were treated with CG+AL(1:4), the expression of mRNA were significantly decreased at the Inflammatory cytokines(IL-1beta, IL-6, TNF-a). While, the levels of mRNA were significantly increased at immune T cell activation cytokines. Additionally, CG+AL(1:4) combinational prescription group enhanced immune cells such as T/B cell and macrophage, NK cell. Furthermore, the Immuno-fluorescence result of brain tissue can confirm that CG+AL(1:4) group significantly increased the BDNF expression. Conclusion : These result suggest that CG+AL(1:4) combinational prescription has Immune System enhancement via stress-mediated immunocyte.

발효 인삼꽃 추출물의 경구 투여가 마우스 사이토카인 IL-6, TNF-α의 생성에 미치는 영향 (Effects of Extract from Fermented Flower-buds of Panax ginseng C.A. Meyer on Mouse Cytokine IL-6, TNF-α Production)

  • 정수지;김경희;손화영;육홍선
    • 한국식품영양학회지
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    • 제27권1호
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    • pp.43-49
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    • 2014
  • 생체 내 실험에서 발효 인삼꽃 추출물(FM), 발효하지 않은 인삼꽃 추출물(FD)과 대조군으로 생리식염수를 2주간 마우스 체중 kg 당 100, 200 mg/kg B.W.의 농도로 마우스에 경구 투여한 후 LPS에 의해 활성화된 복강 대식세포가 분비하는 염증성 사이토카인 IL-6, TNF-${\alpha}$의 생성량을 측정하였다. 그 결과, IL-6는 발효 LPS로 자극한 경우, 두 가지 농도에서 모두 처리한 군에 비해 높은 증식능을 나타내었고, LPS로 자극한 결과, 특히 발효 인삼꽃 추출물 200 mg/kg B.W. 농도에서 유의적으로 낮은 IL-6 분비량을 보였다. TNF-${\alpha}$의 경우, 100 mg/kg B.W.와 200 mg/kg B.W. 두 농도 모두에서 LPS로 자극하지 않은 경우, 낮은 증식 효과를 보여주었고, 자극한 경우, 인삼꽃 시료를 첨가한 군이 대조군보다 낮은 TNF-${\alpha}$ 분비량을 보였으며, FM에서 FD보다 더 TNF-${\alpha}$를 억제하는 효과가 큰 것을 볼 수 있었다. 이상의 결과에 따르면 FD의 사이토카인 IL-6, TNF-${\alpha}$ 생성 효과는 200 mg/kg B.W. 농도 투여 시 효과적으로 면역 세포와 면역 기관의 주요 기능을 증진시킬 가능성이 있을 것으로 사료된다. 이러한 연구결과를 토대로 앞으로 인삼꽃 발효를 이용한 기능성 사료 개발과 더불어 산업적 측면에서 보다 긍정적인 효과를 얻을 수 있을 것으로 판단된다.

옻 추출물의 세포독성 및 자궁 경부암 바이러스 암 유발인자 E6 와 E7의 작용에 미치는 효과 (The Effects of Rhus Extracts on The Cytotoxicity on Cancer Cells and E6 and E7 Oncogenes of Human Papillomavirus Type 16)

  • 조영식;정옥;조정원;이경애;심정현;김광수;이홍수;성기승;윤도영
    • 한국식품과학회지
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    • 제32권6호
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    • pp.1389-1395
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    • 2000
  • 자궁 경부암은 매년 약 50만명 정도씩 사망하는 여성의 치명적인 사망원인의 하나이다. 인두유종 바이러스(HPV) 16형 및 18형과 자궁 경부암과의 긴밀한 관련성은 잘 알려져 있다. 옻 추출물 Rhus가 HPV 16형의 E6, E7 발암 유전자를 억제하는지 여부를 측정하였다. 이 Rhus는 자궁 경부암 세포주(C-33A, SiHa, Caski)와 HaCaT keratinocytes의 분열은 농도 의존적으로 억제하였다. In vitro binding assay와 효소면역측정법에 의하면 Rhus가 암 억제인자인 p53과 결합하여 분해 시키는데 필수적인 E6와 E6AP와의 결합을 억제할 뿐더러 암 억제인자 Rb와 E7과의 결합을 억제하였다. RT-PCR에 의하면 Rhus에 의해 E6 mRNA의 level이 감소하였으나 E7 mRNA는 변하지 않았음을 보여주었다. 이들 결과에 의하면 Rhus가 HPV 16형의 E6와 E7의 발암성을 억제함을 보여 주므로 HPV에 의해 유도된 자궁 경부암의 치료에 유효할 것으로 사료되어 좀 더 자세한 in vitro실험 등이 요구된다.

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Cellular responses to 3D printed dental resins produced using a manufacturer recommended printer versus a third party printer

  • Beatriz Sona Cardoso;Mariana Brito da Cruz;Joana Faria Marques;Joao Carlos Roque;Joao Paulo Martins;Rodrigo Cordeiro Malheiro;Antonio Duarte da Mata
    • The Journal of Advanced Prosthodontics
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    • 제16권2호
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    • pp.126-138
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    • 2024
  • PURPOSE. The aim of this study was to evaluate the influence of different 3D dental resins, using a manufacturer recommended printer and a third-party printer, on cellular responses of human gingival cells. MATERIALS AND METHODS. Three NextDent resins (Denture 3D+, C&B MFH and Crowntec) were used to produce specimens on printers NextDent 5100 (groups ND, NC and NT, respectively) and Phrozen Sonic Mini 4K (groups PD, PC and PT, respectively). Human gingival fibroblasts were cultured and biocompatibility was evaluated on days 1, 3 and 7. IL-6 and IL-8 concentrations were evaluated at 3 days using ELISA. Surface roughness was evaluated by a contact profilometer. SEM and fluorescence micrographs were analyzed at days 1 and 7. Statistical analyses were performed using SPSS and mean differences were tested using ANOVA and post-hoc Tukey tests (P < .05). RESULTS. There was an increase in cellular viability after 7 days in groups PC and PT, when compared to group PD. ND group resulted in higher concentration of IL-6 when compared to PT group. SEM and fluorescence micrographs showed less adhesion and thinner morphology of fibroblasts from group PD. No significant differences were found regarding surface roughness. CONCLUSION. The use of different printers or resins did not seem to influence surface roughness. NextDent 5100 and Phrozen Sonic Mini 4K produced resins with similar cellular responses in human gingival fibroblasts. However, Denture 3D+ resin resulted in significantly lower biocompatibility, when compared to C&B MFH and Crowntec resins. Further testing is required to support its long-term use, required for complete dentures.

탱자 (Poncirus trifoliata)의 lipoprotein lipase 억제메커니즘 (A study of the lipoprotein lipase inhibitory mechanism of Poncirus trifoliata water extracts)

  • 이성미;강윤환;김경곤;김태우;최면
    • Journal of Nutrition and Health
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    • 제48권1호
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    • pp.9-18
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    • 2015
  • 본 연구에는 최근 항비만 소재로 연구되고 있는 건조, 미숙탱자의 물 추출물 (PF-W) 소재를 대상으로 폴리페놀 ($52.15{\pm}4.02mg/g$)과 플라보노이드 ($6.56{\pm}0.47mg/g$) 함량을 측정하고 항산화 활성과 세포독성을 시험한 후, 지방 흡수 제어 가능성을 확인하고자 lipoprotein lipase (LPL)의 억제효능을 배양배지와 세포 내의 LPL 함량, LPL mRNA 발현 그리고 LPL 효소활성측정을 통해 검토하였다. 그 결과 PF-W은 3T3-L1 adipocyte에서 LPL mRNA의 발현과 활성에는 영향이 없었으며, LPL의 분비를 억제하는 것을 알 수 있었다. PF-W의 LPL 분비억제기작을 확인하기 위해 다양한 단백질 이동 관련 유전자의 발현을 확인하였고, 그 결과 LPL의 이동과 분해에 관여하여 세포내 LPL의 활성을 조절하는 것으로 알려진 SorLA의 발현이 증가하는 것을 확인하였다. 이를 조절하는 transcription factor의 발현과 세포핵으로의 이동에 PF-W가 미치는 영향을 검토한 결과 PF-W를 처리함으로써 SorLA promoter 에 작용하는 $C/EBP{\beta}$의 단백질양이 세포핵에서 증가하는 것을 확인할 수 있었다. 본 연구를 통해 PF-W가 SorLA 유전자의 transcription factor인 $C/EBP{\beta}$의 단백질 발현을 세포핵에서 증가시킴으로써 SorLA의 발현이 증가되어 LPL의 분비억제가 가능함을 확인할 수 있었으며 이는 PF-W의 항비만 효과기전을 설명하는 기초자료를 제공하는 것이라 사료된다.

굴 패각 추출물이 Papain으로 유도된 골관절염 C57BL/6J Mice에 미치는 영향 (Effects of Oyster Shell Extract on Papain-induced Osteoarthritis in C57BL/6J Mice)

  • 이세영;김학주;한지숙
    • 한국식품영양과학회지
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    • 제42권8호
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    • pp.1183-1189
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    • 2013
  • 연구에서는 굴 패각 추출물이 papain으로 유도된 골관절염 동물 모델에 미치는 영향을 살펴보았다. 8주령의 C57BL/6J 마우스 우측 슬관절강에 papain $6{\mu}L$를 투여하여 골관절염을 유발하였다. 실험군은 각 군당 체중이 고르게 8마리씩 배정하여 총 다섯군으로 분류하였다. 정상군은 우측 슬관절강에 생리식염수 $6{\mu}L$ 주사하고 20일간 1일 1회 생리식염수 0.2 mL씩 경구투여 하였다. 대조군, 약물투여군, 굴 패각 추출물 투여군은 골관절염을 유발 후 각각 생리식염수, diclofenac sodium 2 mg/kg/bw, OSE 100, 200 mg/kg/bw를 20일 동안 매일 1회 정해진 시간에 경구투여 하였다. 실험기간 동안 체중을 측정하였으며, 실험 20일에 부검하여 슬관절의 병리조직학적 관찰, 관절연골 내 proteoglycan 함유율, 골관절염 지수의 변화, 혈액내 TNF-${\alpha}$, IL-$1{\beta}$, IL-6 함량을 측정하였다. 체중은 실험기간 동안 모든 개체 간의 유의적 차이가 없었다. 관절연골 내 proteoglycan 함유율은 DS 및 OSE 투여군이 papain 투여 대조군보다 유의성(P<0.05)있게 높았으며, 특히 OSE-200 투여군은 약물 투여군과 유사한 proteoglycan 함유율을 나타냈다. H&E 염색을 통하여 관찰한 결과, 골관절염이 유발된 마우스에 굴 패각추출물을 경구 투여하였을 때 관절연골의 파괴와 골 침식 등 연골의 변성이 약물 처리군과 유사한 것을 보아 굴 패각 추출물이 골관절염 치료에 효과가 있는 것을 확인할 수 있었다. 병리조직학적으로 골관절염 지수는 DS 및 OSE 투여군이 papain 투여 대조군보다 유의성(P<0.05) 있게 낮았으며, OSE-200 투여군이 OSE-100 투여군보다 낮았다. 혈액 내 TNF-${\alpha}$, IL-$1{\beta}$, IL-6 함량은 papain 투여 대조군이 정상군에 비해 유의적(P<0.05)으로 증가하였으며, DS 및 OSE 투여군은 대조군보다 유의성(P<0.05) 있게 낮았다. 특히 TNF-${\alpha}$ 및 IL-6 함량은 OSE-200 투여군이 약물 투여군과 유사한 감소 효과를 나타내었다. 이와 같은 결과는 굴 패각 추출물이 papain으로 유도된 골관절염에서 연골의 변성을 줄이고 염증을 억제함으로써 골관절염 치료에 효과가 있는 것으로 사료된다. 굴 패각은 탄산칼슘이 풍부하기 때문에 골관절염 개선에 효과가 있는 것으로 사료되나, 굴 패각 추출물 중 어떠한 기능성 물질이 골관절염 개선 효과를 나타내었는지에 대해 향후 심도 있는 연구가 필요하다.