• IMMUNE NETWORK
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• 제21권6호
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• pp.43.1-43.14
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• 2021

Group 3 innate lymphoid cells (ILC3), which express IL-22 and IL-17A, has been introduced as one of pathologic cells in axial spondyloarthritis (axSpA). Dyslipidaemia should be managed in axSpA patients to reduce cardiovascular disease, and dyslipidaemia promotes inflammation. This study aimed to reveal the role of circulating ILC3 in axSpA and the impact of dyslipidaemia on axSpA pathogenesis. AxSpA patients with or without dyslipidaemia and healthy control were recruited. Peripheral blood samples were collected, and flow cytometry analysis of circulating ILC3 and CD4⁺ T cells was performed. The correlation between Ankylosing Spondylitis Disease Activity Score (ASDAS)-C-reactive protein (CRP) and circulating immune cells was evaluated. The effect of oxidized low-density lipoprotein cholesterol (oxLDL-C) on immune cell differentiation was confirmed. AxSpA human monocytes were cultured with with oxLDL-C, IL-22, or oxLDL-C plus IL-22 to evaluate osteoclastogenesis using tartrate-resistant acid phosphatase (TRAP) staining and real-time quantitative PCR of osteoclast-related gene expression. Total of 34 axSpA patients (13 with dyslipidaemia and 21 without) were included in the analysis. Circulating IL-22⁺ ILC3 and Th17 were significantly elevated in axSpA patients with dyslipidaemia (p=0.001 and p=0.034, respectively), and circulating IL-22⁺ ILC3 significantly correlated with ASDAS-CRP (Rho=0.4198 and p=0.0367). Stimulation with oxLDL-C significantly increased IL-22⁺ ILC3, NKp44^- ILC3, and Th17 cells, and these were reversed by CD36 blocking agent. IL-22 and oxLDL-C increased TRAP⁺ cells and osteoclast-related gene expression. This study suggested potential role of circulating IL-22⁺ ILC3 as biomarker in axSpA. Furthermore, dyslipidaemia augmented IL-22⁺ ILC3 differentiation, and oxLDL-C and IL-22 markedly increased osteoclastogenesis of axSpA.

• 생명과학회지
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• 제18권4호
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• pp.530-536
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• 2008

본 연구는 재조합 단백질 생산에 흔히 사용되는 P. fluorescens에 대한 공기 중 노출 시 염증작용이 의심되는 기전을 분석하기 위하여 인체 폐포 상피세포에서의 염증성 인자들 특히 IL-8, COX-2, MIC-1의 발현을 분석하고자 하였다. 균주 P. fluorescens에 대한 인체 폐포 상피세포 A549에서의 세포증식 억제효과를 밝혔고, 상피세포의 염증성 사이토카인으로서 대표적인 인터루킨 8의 발현에 대해서 분석결과. 균주 P. fluorescens및 재조합 균주에 의해 IL-8의 분비가 진핵세포대비 세균세포숫자 의존적으로 생성량이 증대되었다. 또한 점막 상피세포의 염증성 프로스타글란딘 생성에서 핵심적인 역할을 하는 cyclooxygenase-2 (COX-2)에 대해서 P. fluorescens 는 COX-2의 mRNA 발현이 소량 증진되었으나 실제 단백질의 양 및 전사의 변화는 없었다. 일반적으로 단핵구의 염증성 사이토카인 생성에 대하여 억제성 작용을 하는 macrophage inhibitory cytokine 1 (MIC-1)의 발현에 대해서 측정결과 유전자의 발현이 증대되는 효과는 미비하였으나, MIC-1 단백질의 processing에서 propeptide (${\sim}28\;kD$) 및 mature MIC-1 (${\sim}15\;kD$)로 분해 되어 MIC-1의 단백질 활성화가 증대되었다. 본 연구에서 보이는 MIC-1은 P. fluorescens에 의한 세포 생존율 억제작용에도 기여할 것으로 예측되며, IL-8에 의한 염증구의 recruitment 대한 억제작용이 예상되나, MIC-1의 활성이 과도하고 지속적으로 나타날 경우 조직의 손상도 가능성도 있다.

• Journal of the Korean Wood Science and Technology
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• 제47권6호
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• pp.674-691
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• 2019

본 연구는 국내산 편백, 소나무, 잣나무, 일본잎갈나무 목부 정유의 항염증 효과를 평가 하였다. 실험에 사용된 정유는 각 수종의 목부에서 증류법(hydrodistillation)으로 추출되었다. 추출한 정유들을 GC-MS를 통해 성분 분석한 결과, 편백의 경우 ${\alpha}$-cadinol (19.25%), ${\tau}$-muurolol (14.20%) 및 ${\alpha}$-pinene (13.74%)이, 소나무는 ${\alpha}$-pinene (47.16%), longifolene (14.31%), ${\beta}$-phellandrene (11.78%) 및 ${\beta}$-pinene (11.02%)이 주요성분이었으며, 잣나무의 경우 ${\alpha}$-pinene (13.49%)와 longifolene(10.79%), 낙엽송의 경우 geranyl linalool (23.58%) 및 ${\alpha}$-pinene (18.57%)이 주요성분으로 확인되었다. 침엽수 목부 정유들의 항염증 효과는 LPS(Lipopolysaccharide)로 염증반응이 유도된 RBL-2H3 비만세포주에 정유를 처리한 후 IL-4(interleukin-4), IL-13(interleukin-13), ${\beta}$-hexosaminidase의 발현 변화를 조사함으로써 평가하였다. MTT assay를 통한 세포 독성 평가 결과, 침엽수 4종 정유 모두 $10^{-7}{\sim}10^{-5}%$ 농도에서 세포 독성을 보이지 않았다. 염증반응이 유도된 RBL-2H3 세포에 침엽수 4종의 목부 정유를 $10^{-7}%$ 농도로 처리하였을 때, LPS 처리군 대비 침엽수 4종의 목부 처리군에서 IL-4와 IL-13의 상대적 발현량이 큰 폭으로 감소하였으며, ${\beta}$-hexosaminidase 분비가 유의하게 억제되었다. 침엽수 정유 4종의 IL-4 상대적 발현량은 LPS 처리군 대비 소나무 69.6%, 잣나무 63.2%, 편백 55.1% 그리고 낙엽송 45.8% 감소하였다. IL-13의 상대적 발현량은 소나무 57.8%, 잣나무 57.1%, 편백 51.1% 그리고 낙엽송 34.5% 감소하였다. ${\beta}$-Hexosaminidase 분비의 경우 LPS 처리군 대비 편백 38.1%, 소나무 33.0%, 잣나무 27.4% 그리고 낙엽송 9.1% 억제하였다. Il-4, IL-13 및 ${\beta}$-hexosaminidase 분비를 비교한 결과, 침엽수 목부 정유 4종 중 소나무 정유의 활성이 가장 우수하였다. 이러한 결과는 국내 침엽수 4종의 목부 정유는 염증반응을 유발하는 싸이토카인의 발현 억제를 통하여 항염증 효과를 나타냄을 확인할 수 있었다.

• Tuberculosis and Respiratory Diseases
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• 제80권1호
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• pp.77-82
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• 2017

Background: Delayed hypersensitivity plays a large role in the pathogenesis of tuberculous pleural effusion (TPE). Macrophages infected with live Mycobacterium tuberculosis (MTB) increase the levels of adenosine deaminase2 (ADA2) in the pleural fluid of TPE patients. However, it is as yet unclear whether ADA2 can be produced by macrophages when challenged with MTB antigens alone. This study therefore evaluated the levels of ADA2 mRNA expression, using monocyte-derived macrophages (MDMs) stimulated with MTB antigens. Methods: Purified monocytes from the peripheral blood mononuclear cells of healthy volunteers were differentiated into macrophages using granulocyte-macrophage colony-stimulating factor (GM-CSF) or macrophage colony-stimulating factor (M-CSF). The MDMs were stimulated with early secretory antigenic target protein 6 (ESAT6) and culture filtrate protein 10 (CFP10). The mRNA expression levels for the cat eye syndrome chromosome region, candidate 1 (CECR1) gene encoding ADA2 were then measured. Results: CECR1 mRNA expression levels were significantly higher in MDMs stimulated with ESAT6 and CFP10, than in the unstimulated MDMs. When stimulated with ESAT6, M-CSF-treated MDMs showed more pronounced CECR1 mRNA expression than GM-CSF-treated MDMs. Interferon-${\gamma}$ decreased the ESAT6- and CFP10-induced CECR1 mRNA expression in MDMs. CECR1 mRNA expression levels were positively correlated with mRNA expression of tumor necrosis factor ${\alpha}$ and interleukin 10, respectively. Conclusion: ADA2 mRNA expression increased when MDMs were stimulated with MTB antigens alone. This partly indicates that pleural fluid ADA levels could increase in patients with culture-negative TPE. Our results may be helpful in improving the understanding of TPE pathogenesis.

• 한국식품영양과학회지
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• 제46권3호
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• pp.306-313
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• 2017

본 연구에서는 전통적인 기능성 식품이자 의약품으로 사용되었던 자죽염(PubS)과 복분자를 조합한 복분자 자죽염(PuBS-R)을 사용해 비특이적, 선천성 면역에 중요한 역할을 하는 대식세포에서의 영향을 확인하였다. PuBS-R은 마우스 대식세포인 Raw264.7 세포에서 $1,000{\mu}g/mL$ 농도처리 시 세포의 증식을 유발하였으며, 그에 반해 PuBS를 $1,000{\mu}g/mL$ 처리 시 유의하게 세포 증식률이 억제되었다. 이어서 면역 관련 사이토카인 $TNF-{\alpha}$, $IFN-{\gamma}$, IL-12, IL-10 등을 유전자 단계와 단백질 단계에서 평가하였다. 그 결과 PuBS-R은 50, 100, $500{\mu}g/mL$에서 유의한 증가율을 보였고, 무엇보다 이것은 PuBS 군에 비해서도 월등한 증가율을 확인할 수 있었다. 마지막으로 마우스의 복강에서 분리한 peritoneal macrophage에 PuBS-R을 처리 후 $TNF-{\alpha}$, $IFN-{\gamma}$, IL-12, IL-10, iNOs를 단백질 단계에서 평가한 결과, Raw264.7 세포에서 얻은 결과와 동일한 결과를 확인할 수 있었다. 이러한 결과로 미루어 보아 PuBS-R은 인체의 대식세포 활성의 증가를 통해 비특이적, 선천성 면역력을 증가시킬 것으로 판단되며, 이러한 결과를 바탕으로 추후 실험동물을 이용한 in vivo 후속 연구를 통해 PuBS-R의 면역증강 기능성 식품 개발이 가능할 것으로 생각된다.

• Allergy, Asthma & Immunology Research
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• 제10권6호
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• pp.698-715
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• 2018

Purpose: Hrd1 has recently emerged as a critical regulator of B-cells in autoimmune diseases. However, its role in the pathogenesis of chronic rhinosinusitis with nasal polyps (CRSwNP) remains largely unexplored. This study aimed to examine Hrd1 expression and B-cell accumulation and their possible roles in CRSwNP. Methods: Quantitative real-time polymerase chain reaction, immunohistochemistry, enzyme-linked immunosorbent assay and Western blotting were used to assess gene and protein expression in nasal tissue extracts. Cells isolated from nasal tissues and peripheral blood mononuclear cells were characterized by flow cytometry. Local antibody production was measured in tissue extracts with a Bio-Plex assay. Additionally, changes in Hrd1 expression in response to specific inflammatory stimuli were measured in cultured dispersed polyp cells. Results: Nasal polyps (NPs) from patients with eosinophilic CRSwNP (ECRS) had increased levels of Hrd1, B-cells and plasma cells compared with NPs from patients with non-eosinophilic CRSwNP (non-ECRS) or other control subjects (P < 0.05). The average Hrd1 levels in B-cells in NPs from ECRS patients were significantly higher than those from non-ECRS patients and control subjects (P < 0.05). NPs also contained significantly increased levels of several antibody isotypes compared with normal controls (P < 0.05). Interestingly, Hrd1 expression in cultured polyp cells from ECRS patients, but not non-ECRS patients, was significantly increased by interleukin-$1{\beta}$, lipopolysaccharide and Poly(I:C) stimulation, and inhibited by dexamethasone treatment (P < 0.05). Conclusions: Differential Hrd1 expression and B-cell accumulation between the ECRS and non-ECRS subsets suggests that they can exhibit distinct pathogenic mechanisms and play important roles in NP.

• Clinical and Experimental Otorhinolaryngology
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• 제11권4호
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• pp.281-287
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• 2018

Objectives. Regenerative treatment using stem cells may serve as treatment option for empty nose syndrome (ENS), which is caused by the lack of turbinate tissue and deranged nervous system in the nasal cavity. We aimed to assess the efficacy and safety of the autologous stromal vascular fraction (SVF) in the treatment of ENS. Methods. In this prospective observational clinical study, we enrolled 10 ENS patients who volunteered to undergo treatment of ENS through the injection of autologous SVF. Data, including demographic data, pre- and postoperative Sino-Nasal Outcome Test-25 (SNOT-25) scores, overall patient satisfaction, and postoperative complications, were prospectively collected. Nasal secretion was assessed using the polyurethane foam absorption method, and the levels of biological markers were analyzed in both ENS group and control group using enzyme-linked immunosorbent assay. The SVF extracted from abdominal fat was diluted and injected into both inferior turbinates. Results. Among the 10 initial patients, one was excluded from the study. Subjective satisfaction was rated as "much improved" in two and "no change" in seven. Among the improved patients, the mean preinjection SNOT-25 score was 55.0 and the score at 6 months after injection was 19.5. However, the average SNOT-25 score of nine participants at 6 months after injection (mean${\pm}$standard deviation, $62.4{\pm}35.8$) did not differ significantly from the baseline SNOT-25 score ($70.1{\pm}24.7$, P>0.05, respectively). Among the various inflammatory markers assessed, the levels of interleukin $(IL)-1{\beta}$, IL-8, and calcitonin gene-related peptide were significantly higher in ENS patients. Compared with preinjection secretion level, the nasal secretions from SVF-treated patients showed decreased expressions of $IL-1{\beta}$ and IL-8 after injection. Conclusion. Although SVF treatment appears to decrease the inflammatory cytokine levels in the nasal mucosa, a single SVF injection was not effective in terms of symptom improvement and patient satisfaction. Further trials are needed to identify a more practical and useful regenerative treatment modality for patients with ENS.

• 생명과학회지
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• 제31권10호
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• pp.898-904
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• 2021

본 연구는 lipopolysaccharide (LPS)로 자극된 RAW264.7 세포에 대한 풀무치 유래 항균 펩타이드 locustacin의 항염증 메커니즘을 조사하였다. Locustacin (50, 100, 200 ㎍/ml)은 세포 독성 없이 LPS로 자극된 대식세포의 nitric oxide (NO) 생성을 유의하게 감소시켰고, 단백질과 mRNA 수준에서 inducible NO synthase (iNOS), cyclooxygenase-2 (COX-2)와 같은 전염증 매개체의 발현을 억제하였다. Locustacin은 LPS 처리로 증가된 염증성 사이토카인인 interleukin (IL)-6 및 IL-1β 함량과 이들의 유전자 발현을 모든 처리 농도에서 농도의존적으로 감소시켰다. 한편, LPS에 의해 인산화된 extracellular signal regulated kinase (ERK), p38 및 c-Jun N-terminal kinase (JNK)는 locustacin (100, 200 ㎍/ml) 처리로 억제되었다. 또한, LPS에 의해 유도된 inhibitory kappa B alpha (IκB-α)의 분해를 locustacin이 단백질 수준에서 억제한다는 것을 발견했다. 결론적으로, locustacin은 LPS 처리된 대식세포에서 mitogen-activated protein kinases (MAPKs) 인산화, nuclear factor kappa B (NF-κB) 활성화 및 하위 염증 매개체를 억제함으로써 항염증 효과를 가지고 있음을 확인하였다. 이러한 결과들은 풀무치 전사체 분석을 통해 확인된 locustacin이 항염증제 후보물질로서 개발 가능성이 있음을 제시한다.

• Journal of Ginseng Research
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• 제39권3호
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• pp.230-237
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• 2015

Background: Colorectal cancer (CRC) is a leading cause of death worldwide. Chronic gut inflammation is recognized as a risk factor for tumor development, including CRC. American ginseng is a very commonly used ginseng species in the West. Methods: A genetically engineered $Apc^{Min/+}$ mouse model was used in this study. We analyzed the saponin composition of American ginseng used in this project, and evaluated its effects on the progression of high-fat-diet-enhanced CRC carcinogenesis. Results: After oral ginseng administration (10-20 mg/kg/d for up to 32 wk), experimental data showed that, compared with the untreated mice, ginseng very significantly reduced tumor initiation and progression in both the small intestine (including the proximal end, middle end, and distal end) and the colon (all p < 0.01). This tumor number reduction was more obvious in those mice treated with a low dose of ginseng. The tumor multiplicity data were supported by body weight changes and gut tissue histology examinations. In addition, quantitative real-time polymerase chain reaction analysis showed that compared with the untreated group, ginseng very significantly reduced the gene expression of inflammatory cytokines, including interleukin-$1{\alpha}$ (IL-$1{\alpha}$), IL-$1{\beta}$, IL-6, tumor necrosis factor-${\alpha}$, granulocyte-colony stimulating factor, and granulocyte-macrophage colony-stimulating factor in both the small intestine and the colon (all p < 0.01). Conclusion: Further studies are needed to link our observed effects to the actions of the gut microbiome in converting the parent ginsenosides to bioactive ginseng metabolites. Our data suggest that American ginseng may have potential value in CRC chemoprevention.

• Tuberculosis and Respiratory Diseases
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• 제82권2호
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• pp.133-142
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• 2019

Background: Idiopathic pulmonary fibrosis involves irreversible alveolar destruction. Although alveolar epithelial type II cells are key functional participants within the lung parenchyma, how epithelial cells are affected upon bleomycin (BLM) exposure remains unknown. In this study, we determined whether BLM could induce cell cycle arrest via regulation of Schlafen (SLFN) family genes, a group of cell cycle regulators known to mediate growth-inhibitory responses and apoptosis in alveolar epithelial type II cells. Methods: Mouse AE II cell line MLE-12 were exposed to $1-10{\mu}g/mL$ BLM and $0.01-100{\mu}M$ baicalein (Bai), a G1/G2 cell cycle inhibitor, for 24 hours. Cell viability and levels of pro-inflammatory cytokines were analyzed by MTT and enzyme-linked immunosorbent assay, respectively. Apoptosis-related gene expression was evaluated by quantitative real-time reverse transcription-polymerase chain reaction (qRT-PCR). Cellular morphology was determined after DAPI and Hoechst 33258 staining. To verify cell cycle arrest, propidium iodide (PI) staining was performed for MLE-12 after exposure to BLM. Results: BLM decreased the proliferation of MLE-12 cells. However, it significantly increased expression levels of interleukin 6, tumor necrosis factor ${\alpha}$, and transforming growth factor ${\beta}1$. Based on Hoechst 33258 staining, BLM induced condensation of nuclear and fragmentation. Based on DAPI and PI staining, BLM significantly increased the size of nuclei and induced G2/M phase cell cycle arrest. Results of qRT-PCR analysis revealed that BLM increased mRNA levels of BAX but decreased those of Bcl2. In addition, BLM/Bai increased mRNA levels of p53, p21, SLFN1, 2, 4 of Schlafen family. Conclusion: BLM exposure affects pulmonary epithelial type II cells, resulting in decreased proliferation possibly through apoptotic and cell cycle arrest associated signaling.