• 생명과학회지
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• 제25권1호
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• pp.84-92
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• 2015

Interferon inducible transmembrane protein (IFITM) family 유전자는 인터페론(IFNs)의 동형 세포부착 기능 및 세포의 항-증식 활성과 같은 몇 가지 세포증식 과정에 연관되어 있다. 본 연구에서는 IFITM2 및 IFITM5 SNPs이 궤양성대장염의 감수성과 연관되어 있는지 알아 보고자 했다. 본 연구에서 직접 염기서열 분석법을 사용하여 IFITM2 유전자에서 총 13개, IFITM5 유전자에서는 12개의 유전적 변이를 발굴하였다. 이들의 SNPs의 유전자형 분석은 PCR-RFLP 법과 Taq-Man probe 분석법을 사용하였고, 일배체형 빈도 분석은 EM algorithm을 사용하여 분석하였다. 궤양성대장염 환자에서 IFITM2 및 IFITM5 SNPs의 유전자형과 대립유전자 빈도는 건강인 대조군과 비교했을 때 유의성이 없었다. 궤양성대장염 환자와 정상인 대조군에서 IFITM1의 rs77537847, IFITM2의 rs909097, IFITM5의 rs56069858을 지표로 하는 유전자형 조합 빈도를 분석한 결과 주된 유전자형 조합빈도에서는 유의성이 없는 것으로 나타났으나, 궤양성대장염 환자와 건강인 대조군의 GGT 유전자형조합 빈도 분석에서는 유의하게 다른 차이를 보였다(p=0.002). 이러한 결과에 의거하여 IFITMs의 SNPs 유전자형 조합이 궤양성대장염의 감수성과 연관성이 있고, 궤양성대장염의 유용한 유전자 마커로 사용 할 수 있다고 생각된다.

• Molecules and Cells
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• 제38권11호
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• pp.998-1006
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• 2015

Retinols are metabolized into retinoic acids by alcohol dehydrogenase (ADH) and retinaldehyde dehydrogenase (Raldh). However, their roles have yet to be clarified in hepatitis despite enriched retinols in hepatic stellate cells (HSCs). Therefore, we investigated the effects of retinols on Concanavalin A (Con A)-mediated hepatitis. Con A was injected into wild type (WT), Raldh1 knockout ($Raldh1^{-/-}$), $CCL2^{-/-}$ and $CCR2^{-/-}$ mice. For migration study of regulatory T cells (Tregs), we used in vivo and ex vivo adoptive transfer systems. Blockade of retinol metabolism in mice given 4-methylpyrazole, an inhibitor of ADH, and ablated Raldh1 gene manifested increased migration of Tregs, eventually protected against Con A-mediated hepatitis by decreasing interferon-${\gamma}$ in T cells. Moreover, interferon-${\gamma}$ treatment increased the expression of ADH3 and Raldh1, but it suppressed that of CCL2 and IL-6 in HSCs. However, the expression of CCL2 and IL-6 was inversely increased upon the pharmacologic or genetic ablation of ADH3 and Raldh1 in HSCs. Indeed, IL-6 treatment increased CCR2 expression of Tregs. In migration assay, ablated CCR2 in Tregs showed reduced migration to HSCs. In adoptive transfer of Tregs in vivo and ex vivo, Raldh1-deficient mice showed more increased migration of Tregs than WT mice. Furthermore, inhibited retinol metabolism increased survival rate (75%) compared with that of the controls (25%) in Con A-induced hepatitis. These results suggest that blockade of retinol metabolism protects against acute liver injury by increased Treg migration, and it may represent a novel therapeutic strategy to control T cell-mediated acute hepatitis.

• 대한화장품학회지
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• 제36권2호
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• pp.151-156
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• 2010

버섯유래 생리활성물질을 탐색하고자, 젖버섯아재비 자실체로부터 각종 컬럼크로마토그래피 및 HPLC 등 기법에 의하여 4종의 azulene계 화합물을 순수히 분리정제 하였다. 분리된 화합물은 각종 물리화학적 특성 및 분광학적 분석 자료에 의하여 1-formyl-4-methyl-7-isopropyl azulene (1), lactaroviolin (2), 4-methyl-7-isopropyl-azulene-1-carboxylic acid (3) 및 1-formyl-4-methyl-7-(1-hydroxy-1-methylethyl) azulene (4)로 동정되었다. 이들 화합물의 인터페론 감마 생성에 미치는 영향을 조사하였다. 화합물 1과 4는 자연살해세포주(NK92 cell)에서 인터페론 감마 생성을 농도 의존적으로 억제하였으며, 400 ${\mu}M$농도에서 각각 101.3 %와 92.7 %, 100 ${\mu}M$농도에서 각각 11.9 %와 24.1 %의 높은 저해활성을 보였으며, 화합물 2와 3은 400 ${\mu}M$농도에서 45.9 %와 18.0 %의 다소 낮은 저해활성을 나타내었다.

• Journal of Ginseng Research
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• 제41권1호
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• pp.43-51
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• 2017

Background: BIOGF1K, a compound K-rich fraction prepared from the root of Panax ginseng, is widely used for cosmetic purposes in Korea. We investigated the functional mechanisms of the anti-inflammatory and antioxidative activities of BIOGF1K by discovering target enzymes through various molecular studies. Methods: We explored the inhibitory mechanisms of BIOGF1K using lipopolysaccharide-mediated inflammatory responses, reporter gene assays involving overexpression of toll-like receptor adaptor molecules, and immunoblotting analysis. We used the 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay to measure the antioxidative activity. We cotransfected adaptor molecules, including the myeloid differentiation primary response gene 88 (MyD88) and Toll/interleukin-receptor domain containing adaptor molecule-inducing interferon-${\beta}$ (TRIF), to measure the activation of nuclear factor (NF)-${\kappa}B$ and interferon regulatory factor 3 (IRF3). Results: BIOGF1K suppressed lipopolysaccharide-triggered NO release in macrophages as well as DPPH-induced electron-donating activity. It also blocked lipopolysaccharide-induced mRNA levels of interferon-${\beta}$ and inducible nitric oxide synthase. Moreover, BIOGF1K diminished the translocation and activation of IRF3 and NF-${\kappa}B$ (p50 and p65). This extract inhibited the upregulation of NF-${\kappa}B$-linked luciferase activity provoked by phorbal-12-myristate-13 acetate as well as MyD88, TRIF, and inhibitor of ${\kappa}B$ ($I{\kappa}B{\alpha}$) kinase ($IKK{\beta}$), and IRF3-mediated luciferase activity induced by TRIF and TANK-binding kinase 1 (TBK1). Finally, BIOGF1K downregulated the NF-${\kappa}B$ pathway by blocking $IKK{\beta}$ and the IRF3 pathway by inhibiting TBK1, according to reporter gene assays, immunoblotting analysis, and an AKT/$IKK{\beta}$/TBK1 overexpression strategy. Conclusion: Overall, our data suggest that the suppression of $IKK{\beta}$ and TBK1, which mediate transcriptional regulation of NF-${\kappa}B$ and IRF3, respectively, may contribute to the broad-spectrum inhibitory activity of BIOGF1K.

• 대한수의학회지
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• 제51권2호
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• pp.117-122
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• 2011

Bovine tuberculosis (bTB), caused primarily by Mycobacterium bovis, continues to exert an economic loss, even in countries with active control measures, and is one of zoonotic diseases enable to be transmitted to human. The control and eradication of bTB are mainly based on a test and slaughter policy and/or abattoir surveillance. Various factors including limitation of diagnostic tests have been considered as major constraints to eradication. Single intradermal test (SIT) is the official diagnostic test. New diagnostic methods are needed to be developed, because of limitations of the test. In the present study SIT was compared with single intradermal comparative cervical test (SICCT) and interferon (IFN)-${\gamma}$ assay. There was very low correlation between SIT and SICCT. However, high correlation was shown between SIT and IFN-${\gamma}$ assay while no correlation was observed between SICCT and IFN-${\gamma}$ assay. Therefore, our results suggest the possibility of replacement of SIT with IFN-${\gamma}$ assay for the diagnosis of bovine tuberculosis.

• BMB Reports
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• 제52권5호
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• pp.318-323
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• 2019

Mesenchymal stem cells (MSCs) are multipotent adult stem cells that present immunosuppressive effects in experimental and clinical trials targeting various rare diseases including inflammatory bowel disease (IBD). In addition, recent studies have reported tryptophanyl-tRNA synthetase (WRS) possesses uncanonical roles such as angiostatic and anti-inflammatory effects. However, little is known about the function of WRS in MSC-based therapy. In this study, we investigated if a novel factor, WRS, secreted from MSCs has a role in amelioration of IBD symptoms and determined a specific mechanism underlying MSC therapy. Experimental colitis was induced by administration of 3% DSS solution to 8-week-old mice and human umbilical cord blood-derived MSCs (hUCB-MSCs) were injected intraperitoneally. Secretion of WRS from hUCB-MSCs and direct effect of WRS on isolated $CD4^+$ T cells was determined via in vitro experiments and hUCB-MSCs showed significant therapeutic rescue against experimental colitis. Importantly, WRS level in serum of colitis induced mice decreased and recovered by administration of MSCs. Through in vitro examination, WRS expression of hUCB-MSCs increased when cells were treated with interferon-${\gamma}$ ($IFN-{\gamma}$). WRS was evaluated and revealed to have a role in inhibiting activated T cells by inducing apoptosis. In summary, $IFN-{\gamma}$-mediated secretion of WRS from MSCs has a role in suppressive effect on excessive inflammation and disease progression of IBD and brings new highlights in the immunomodulatory potency of hUCB-MSCs.

• 한국동물위생학회지
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• 제43권4호
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• pp.201-209
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• 2020

In Korea, bovine tuberculosis (bTB) is a representative zoonotic disease that causes considerable economic loss. In determining the positive bTB, the ELISA method for examining the amount of interferon-gamma (IFN-γ) is included in Korea's diagnostic standard method. Recently, commercially available BIONOTE TB-Feron ELISA Plus (TB-Feron Plus) that detects IFN-γ has been introduced. However, since the scientific basis for the performance is limited, we evaluated performance by comparing it with the results of another IFN-γ ELISA assay kit (BOVIGAM®) certified by Office International des Epizooties. In our research, 42 positive blood samples preliminarily tested with a tuberculin skin test and/or BOVIGAM® and 54 negative blood samples collected from three bTB free farms were subjected to IFN-γ assay using the TB-Feron Plus and the BOVIGAM®, respectively. The result shows that the sensitivity, specificity and accuracy were 81.0% (34/42), 100% (54/54), 91.7% (88/96) in TB-Feron Plus kit and 78.6% (33/42), 100% (54/54), 90.6% (87/96) in BOVIGAM® kit, respectively. Moreover, the overall accordance percentage of the two kits was 99.0% (95/96) and there was almost perfect agreement between two assays (Kappa=0.977, P<0.0001). Furthermore, additional studies confirmed that elevated lymphocyte numbers in blood did not interfere with the results of the TB-Feron Plus kit. And, delayed time from sampling to culture decreased the optical density (OD) value. Therefore, we concluded that the TB-Feron Plus kit was not inferior to BOVIGAM® in performance. High lymphocyte numbers in blood did not impact on TB-Feron Plus results, while delayed time before culture interfered with OD value.

• Animal Bioscience
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• 제36권6호
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• pp.851-860
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• 2023

Objective: This study aims to evaluate the target genes of gga-miR-20a-5p and the regulated immune responses in the chicken macrophage cell line, HD11, by the exosome-mediated delivery of miR-20a-5p. Methods: Exosomes were purified from the chicken macrophage cell line HD11. Then, mimic gga-miR-20p or negative control miRNA were internalized into HD11 exosomes. HD11 cells were transfected with gga-miR-20a-5p or negative control miRNA containing exosomes. After 44 h of transfection, cells were incubated with or without 5 ㎍/mL poly(I:C) for 4 h. Then, expression of target genes and cytokines was evaluated by quantitative realtime polymerase chain reaction. Results: Using a luciferase reporter assay, we identified that gga-miR-20a-5p directly targeted interferon gamma receptor 2 (IFNGR2), mitogen-activated protein kinase 1 (MAPK1), mitogen-activated protein kinase kinase kinase 5 (MAP3K5), and mitogen-activated protein kinase kinase kinase 14 (MAP3K14). Moreover, the exosome-mediated delivery of gga-miR-20a-5p successfully repressed the expression of IFNGR2, MAPK1, MAP3K5, and MAP3K14 in HD11 cells. The expressions of interferon-stimulated genes (MX dynamin like GTPase 1 [MX1], eukaryotic translation initiation factor 2A [EIF2A], and oligoadenylate synthase-like [OASL]) and proinflammatory cytokines (interferon-gamma [IFNG], interleukin-1 beta [IL1B], and tumor necrosis factor-alpha [TNFA]) were also downregulated by exosomal miR-20a-5p. In addition, the proliferation of HD11 cells was increased by exosomal miR-20a-5p. Conclusion: The exosome-mediated delivery of gga-miR-20a-5p regulated immune responses by controlling the MAPK and apoptotic signaling pathways. Furthermore, we expected that exosomal miR-20a-5p could maintain immune homeostasis against highly pathogenic avian influenza virus H5N1 infection by regulating the expression of proinflammatory cytokines and cell death.

• 생명과학회지
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• 제34권6호
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• pp.408-417
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• 2024

원화(Daphne genkwa (Siebold & Zucc))의 50% 에탄올 추출물인 PB-81이 소 로타바이러스 설사병 환축에서 설사병의 치료효과와 바이러스 증식 억제 효과가 나타나는가를 확인하였다. PB-81에 의해 상피세포주인 A549 세포와 혈액세포주인 NK92 세포에서 각각 IFN-β와 IFN-γ가 유도되는 것을 확인하였다. PB-81의 바이러스의 증식억제 효과를 확인하기 위해 PB-81을 MBDK 세포에 바이러스의 감염 전, 동시, 감염 후에 투여하는 세 가지 경우에서 바이러스 억제효과를 확인한 결과 PB-81을 투여한 모든 경우에서 바이러스가 억제되었으며, 바이러스 감염 전에서 투여하는 경우에서 가장 좋은 바이러스 억제효과가 나타났다. 마우스에서의 독성검사에서는 투여최대용량인 20 mg/mL에서도 독성에 따른 부작용이 나타나지 않았다. 소 로타바이러스 설사병 환축 16두와 대조군 4두를 대상으로 한 치료효과 검증에는 20 mg/5 mL 용량의 PB-81을 투여한 결과, PB-81을 투여한 모든 소 로타바이러스 설사병 환축이 완치되었으며 PB-81의 투여 후에 완치까지 소요된 기간은 PB-81투여군이 평균 2.25일이었고, PB-81을 투여하지 않은 대조군이 6.5일로 PB-81의 투여에 의한 소 로타바이러스 설사병의 치료효과가 나타나는 것을 확인하였으며 PB-81을 투여한 모든 환축에서 부작용이 검출되지 않았다.

• 한국미생물학회:학술대회논문집
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• 한국미생물학회 2005년도 International Meeting of the Microbiological Society of Korea
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• pp.228.3-228
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• 2005