• Journal of Microbiology and Biotechnology
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• 제22권4호
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• pp.494-500
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• 2012

Silk fibroin (SF) peptide has been traditionally used as a treatment for flatulence, spasms, and phlegm. In this study, we examined whether SF peptide enhanced the anti-inflammatory effect of PEP-1-FK506 binding protein (PEP-1-FK506BP) through comparing the anti-inflammatory activities of SF peptide and/or PEP-1-FK506BP. In the presence or absence of SF peptide, transduction levels of PEP-1-FK506BP into HaCaT cells and mice skin and anti-inflammatory activities of PEP-1-FK506BP were identified by Western blot and histological analyses. SF peptide alone effectively reduced both mice ear edema and the elevated levels of cyclooxygenase-2, interleukin-6 and $-1{\beta}$, and tumor necrosis factor-${\alpha}$, showing similar anti-inflammatory effect to that of PEP-1-FK506BP. Furthermore, co-treatment with SF peptide and PEP-1-FK506BP exhibited more enhanced anti-inflammatory effects than the samples treated with SF peptides or PEP-1-FK506BP alone, suggesting the possibility that SF peptide and PEP-1-FK506BP might interact with each other. Moreover, the transduction data demonstrated that SF peptide did not affect the transduction of PEP-1-FK506BP into HaCaT cells and mice skin, indicating that the improvement of anti-inflammatory effect of PEP-1-FK506BP was not caused by enhanced transduction of PEP-1-FK506BP. Thus, these results suggest the possibility that co-treatment with SF peptide and PEP-1-FK506BP may be exploited as a useful therapy for various inflammation-related diseases.

• Journal of Periodontal and Implant Science
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• 제34권3호
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• pp.607-622
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• 2004

Recently epidemiologic studies have indicated that the patients with periodontitis may have increased risk of ischemic cardiovascular events, and have suggested the important roles of blood cytokines and acute reactant proteins in the systemic infection and inflammatory response. Periodontitis and coronary heart disease (CHD) may share the common risk factors and the genetic mechanism associated with interleukin(IL)-1A, B and RA genotype may be involved in the production of IL-1. This study was aimed to investigate the relationship between angiographically defined CHD and periodontitis as chronic Gram-negative bacterial infection and to determine whether the IL-1 gene polymorphism is associated in both diseases. Patients under the age of 60 who had undergone diagnostic coronary angiography were enrolled in this study. Subjects were classified as positive CHD (+CHD, n=37) with coronary artery stenosis more than 50% in at least one of major epicardial arteries, and negative CHD (-CHD, n=30) without significant stenosis. After recording the number of missing teeth, periodontal disease severity was measured by means of plaque index (PI), gingival index (GI), bleeding on probing (BOP), probing depth (PD), clinical attachment level (CAL), and radiographic bone loss around all remaining teeth. Gingival crevicular fluid (GCF) was collected from the 4 deepest periodontal pockets and assessed for cytokine ($IL-1{\beta}$, IL-6, IL-1ra, tumor necrosis $factor-{\alpha}$, and prostaglandin $E_2$). Additionally, blood CHD markers, lipid profile, and blood cytokines were analyzed. IL-1 gene cluster genotyping was performed by polymerase chain reaction and enzyme restriction using genomic DNA from buccal swab, and allele 2 frequencies of IL-1A(+4845), IL-1B(+3954), IL-B(-511), and IL-1RA(intron 2) were compared between groups. Even though there was no significant difference in the periodontal parameters between 2 groups, GCF level of $PGE_2$ was significantly higher in the +CHD group(p<0.05). Correlation analysis showed the positive relationship among PD, CAL and coronary artery stenosis(%) and blood $PGE_2$. There was also significant positive relationship between the periodontal parameters (PI, PD, CAL) and the blood CHD markers (leukocyte count, C-reactive protein, and lactic dehyrogenase). IL-1 gene genotyping showed that IL-1A(+3954) allele 2 frequency was significantly higher in the +CHD group compared with the -CHD group (15% vs. 3.3%, OR 5.118,p=0.043). These results suggested that periodontal inflammation is related to systemic blood cytokine and CHD markers, and contributes to cardiovascular disease via systemic inflammatory reaction. IL-1 gene polymorphism might have an influence on periodontal and coronary heart diseases in Korean patients.

• 한국식품영양과학회지
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• 제46권6호
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• pp.659-670
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• 2017

산화스트레스와 염증은 간 손상의 진행과정에 중요한 인자로 작용한다. 굴가수분해물의 항산화 및 항염증 활성은 지질대사, 혈압 및 혈당, 면역기능의 조절과 같은 다양한 기능에 관여한다. 그러나 급성 간 손상 모델에서 굴가수분해물의 효과를 확인한 연구 결과는 아직 확인된 바 없다. 본 연구는 LPS/D-GalN에 의해 유도된 급성 간 손상 생쥐 모델에서 굴가수분해물의 효과를 확인하기 위해 수행되었다. 실험군은 대조군(생리식염수), LPS/D-GalN 간 손상군, LPS/D-GalN과 굴가수분해물(100 mg/kg, 200 mg/kg, 400 mg/kg)의 병합투여군 및 LPS/D-GalN과 silymarin(25 mg/kg) 병합투여군으로 나누었다. 급성 간 손상 모델은 $1{\mu}g/kg$의 LPS와 400 mg/kg의 D-GalN으로 유도되었다. 먼저 시료의 항산화 및 항염증 활성을 분석한 결과 굴가수분해물은 농도 의존적으로 높은 DPPH 및 ABTS 라디칼 소거 활성을 보였으며, 인간 정상 간세포주(Chang)에서 과산화수소에 의한 세포 내 활성산소의 생성을 유의적으로 감소시켰다. 또한, 굴가수분해물은 농도 의존적으로 높은 COX-2 및 5-LOX 억제능을 보였으며, LPS에 의해 활성화된 생쥐 대식세포주 RAW264.7에서 발현되는 $TNF-{\alpha}$, IL-6 및 $IL-1{\beta}$의 염증성 사이토카인의 mRNA 발현률을 감소시켰다. 굴가수분해물 투여는 LPS/D-GalN에 의한 혈청 ALT 및 AST 증가를 유의적으로 감소시켰으며, 간 조직의 출혈 및 간세포의 자멸사를 감소시켰다. 또한, 간 균질의 $TNF-{\alpha}$, $IL-1{\beta}$ 및 IL-6 함량을 감소시켰으며, 감소한 catalase의 활성을 유의적으로 증가시켰다. 이상의 결과로부터 굴가수분해물은 간 보호 효과를 가지는 것으로 판단되며, 급성 간 손상의 예방 및 치료에 도움이 될 수 있는 시료로 활용될 수 있을 것으로 기대된다.

• 대한한의학방제학회지
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• 제15권1호
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• pp.163-173
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• 2007

Jeungaektang (JAT) is the herbal formula, has the effect of moistening the dryness by activating lung Qi and by nourishing Yin, has being used for dryness syndromes. Generally the herbal formulae for moistening dryness are used for exogenous or endogenous dryness syndromes. JAT has been clinically used for the treatment of endogenous dryness syndromes. It is composed of Scrophulariae Radix. Rehmanniae Radix and Liriopis Tuber. Recent studies showed that JAT has a protective effect against $CCl_{4}-induced$ hepatotoxicity and anti-inflammatory effects against ear swelling of mouse induced by Crotonis Fructus. However, the effect of JAT on the immunological activity was rarely studied. Therefore, this study evaluated the effects of JAT the regulatory mechanism of nitric oxide (NO) and cytokines in the lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. After the treatment of JAT water extract, cell viability was measured by MTT assay, NO production was monitored by measuring the nitrite content in culture medium. Cyclooxygenase-2 (COX -2) and inducible nitric oxide synthase (iNOS) were determined by immunoblot analysis, and levels of cytokine were analyzed by sandwich immunoassays. Results provided evidence that JAT inhibited the production of nitrite and nitrate ($0.1{\sim}1.0$ mg/ml), iNOS ($0.1{\sim}1.0$ mg/ml), $interleukin-1{\beta}$ ( $0.1{\sim}1.0$ mg/ml) and tumor necrosis $factor-{\alpha}$ ($0.1{\sim}1.0$ mg/ml) in RAW 264.7 cells activated with LPS. Furthermore, JAT inhibited the expression of COX-2 expression and production of prostagladin E2 ($0.1{\sim}1.0$ mg/ml). These findings suggest that JAT can produce anti-inflammatory effect, which may play a role in adjunctive therapy in Gram-negative bacterial infections.

• BMB Reports
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• 제44권5호
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• pp.329-334
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• 2011

Many proteins with poor transduction efficiency were reported to be delivered to cells by fusion with protein transduction domains (PTDs). In this study, we investigated the effect of levosulpiride on the transduction of PEP-1 ribosomal protein S3 (PEP-1-rpS3), and examined its influence on the stimulation of the therapeutic properties of PEP-1-rpS3. PEP-1-rpS3 transduction into HaCaT human keratinocytes and mouse skin was stimulated by levosulpiride in a manner that did not directly affect the cell viability. Following 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced inflammation in mice, levosulpiride alone was ineffective in reducing TPA-induced edema and in inhibiting the elevated productions of inflammatory mediators and cytokines, such as cyclooxygenase-2, inducible nitric oxide synthase, interleukin-6 and -1${\beta}$, and tumor necrosis factor-${\alpha}$. Anti-inflammatory activity by PEP-1-rpS3 + levosulpiride was significantly more potent than by PEP-1-rpS3 alone. These results suggest that levosulpiride may be useful for enhancing the therapeutic effect of PEP-1-rpS3 against various inflammatory diseases.

• 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.

• Tuberculosis and Respiratory Diseases
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• 제52권4호
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• pp.355-366
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• 2002

연구배경: 열 전처치는 조직 내에 열충격단백질의 생성을 유도하며 이러한 열 전처치가 내독소로 유도된 쥐의 급성폐손상을 감소시키고 패혈증에 의한 사망률을 감소시킨다고 알려져 있다. 그러나, 폐손상을 유발하는 원인에 노출된 후 가해진 열처치가 폐손상에 미치는 효과에 대하여는 아직까지 잘 알려져 있지 않다. 따라서 본 연구는 내독소 투여 직후 시행한 열충격이 내독소에 의해 유발된 쥐의 급성폐손상에 미치는 영향과 그에 따른 염증성 및 항염증성 사이토카인에 미치는 영항을 알아보고자 하였다. 방 법: 대조군은 백서의 기관지 내로 생리식염수를 투여하였고 열처치 대조군은 생리식염수 투여 직후 열처치를 시행하였다. 내독소군은 열처치 없이 내독소를 기관지내로 투여하였다. 열 전처치군은 내독소 투여 18시간 전에 열 전처치를 시행하였고, 열 동시처치 군은 내독소 투여 직후 열처치를 시행하였다. 내독소 투여 후 6시간에 기관지폐포세척을 시행하여 기관지폐포세척액 내의 호중구 백분율을 측정하였고 폐를 적출하여 myeloperoxidase(MPO)의 활성도를 측정하였으며 기관지폐포세척액과 혈청에서 LDH(lactic dehydrogenase), 단백질, IL(interleukin)-$1{\beta}$, TNF(tumor necrosis factor)-${\alpha}$ 및 IL-10를 측정하였다. 또한 각군에서 폐 조직 내 HSP72의 표현정도를 관찰하였다. 결 과: 1) 내독소군, 열 전처치군 및 열 동시처치군 모두에서 기관지폐포세척액 내의 호중구 백분율, 폐조직이 MPO, 혈청 및 기관지폐포세척액내의 $IL-1{\beta}$, $TNF-{\alpha}$ 및 IL-10이 대조군에 비해 증가하였다(각 p<0.05). 2) 열 동시처치군은 폐조직의 MPO와 기관지폐포세척액 단백질의 농도가 내독소군과 차이가 없었고 기관지폐포세척액의 LDH가 내독소군에 비해 증가하였다(p<0.05). 3) 열 동시처치군의 혈청 $TNF-{\alpha}$의 농도는 내독소군과 비교 시 증가하였다(p=0.01). 결 론: 내독소 투여 직후 시행한 열충격은 내독소로 유도되는 폐손상을 감소시키지 못하며 염증성 사이토카인의 농도를 증가시켰다.

• Clinical and Experimental Reproductive Medicine
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• 제38권4호
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• pp.193-202
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• 2011

Objective: We found previously that $interferon$ $regulatory$ factor ($Irf$)-1 is a germinal vesicle (GV)-selective gene that highly expressed in GV as compared to metaphase II oocytes. To our knowledge, the function of $Irf-1$ in oocytes has yet to be examined. The present study was conducted to determine the relationship between retinoic acid (RA) and RA-mediated expression of $Irf-1$ and the mouse oocyte maturation. Methods: Immature cumulus-oocyte-complexes (COCs) were collected from 17-day-old female mice and cultured $in$ $vitro$ for 16 hours in the presence of varying concentrations of RA (0-10 ${\mu}M$). Rate of oocyte maturation and activation was measured. Gene expression was measured by quantitative real-time reverse transcription-polymerase chain reaction (RT-PCR) and cytokine secretion in the medium was measured by Bio-Plex analysis. Apoptosis was analyzed by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay. Results: The rates of oocyte maturation to metaphase II and oocyte activation increased significantly with RA treatment (10 nM-1 ${\mu}M$). With 100 nM RA treatment, lowest level of $Irf-1$ mRNA and cumulus cell's apoptosis was found. Among 23 cytokines measured by Bio-Plex system, the substantial changes in secretion of tumor necrosis factor-${\alpha}$, macrophage inflammatory protein-$1{\beta}$, eotaxin and interleukin-12 (p40) from COCs in response to RA were detected. Conclusion: We concluded that the maturation of oocytes and $Irf-1$ expression are negatively correlated, and RA enhances the developmental competence of mouse immature oocytes $in$ $vitro$ by suppressing apoptosis of cumulus cells. Using a mouse model, results of the present study provide insights into improved culture conditions for $in$ $vitro$ oocyte maturation and relevant cytokine production and secretion in assisted reproductive technology.

• 한국식품영양과학회지
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• 제44권10호
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• pp.1450-1457
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• 2015

톳은 여러 생리활성이 알려져 있는 해조류로 본 연구에서는 톳의 활용 가능성을 확장시키기 위하여 유산균의 종류를 달리하여 발효한 톳을 시료로 하고 추출물 단계에서 항염증 효과를 비교하였다. 유산균인 Weissella sp. SH-1과 Lactobacillus casei를 접종하여 발효시킨 톳 추출물은 무접종군에 비하여 높은 NO 억제 활성을 나타내었으며, 유산균간의 비교에서는 Weissella sp. SH-1 접종군보다 L. casei 접종군에서 NO 생성 억제 효과가 높게 나타났다. 중요 염증 유발인자인 iNOS, COX-2 및 IL-6의 발현을 비교한 결과 Weissella sp. SH-1 접종군에 의한 iNOS 억제능이 높았으며 COX-2, IL-6 발현은 L. casei 접종군에 의해 효과적으로 억제되었다. 유산균에 의한 염증 유발인자 억제능에 대한 MAPK 신호 전달 경로를 알아본 결과 ERK, p38, JNK의 인산화에 의해 항염증 활성이 나타나는 것을 알 수 있었다. 이상의 결과로부터 유산균 Weissella sp. SH-1과 L. casei를 이용한 발효는 염증 억제에 효과가 있는 유효성분의 추출을 증진시킬 수 있음을 간접적으로 확인할 수 있었다. 향후 본 연구 결과를 바탕으로 유산균 Weissella sp. SH-1과 L. casei를 이용한 발효방법을 활용하여 기능성 식품소재 및 제품 개발에 응용이 가능할 것으로 기대된다.

• 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.