• 생명과학회지
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• 제25권9호
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• pp.993-999
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• 2015

본 연구에서는 참콩풍뎅이(Popillia flavosellata) 에탄올 추출물(PFE)의 항염증 효능을 분석하기 위해 PFE를 농도별(500, 1,000, 2,000 μg/ml)로 대식세포인 RAW 264.7에 처리 시 최고 처리농도인 2,000 μg/ml까지 통계적인 유의성 있는 독성이 없음을 확인하였다. LPS (100 ng/ml)로 염증 유도된 RAW 264.7 세포에 PFE를 농도별(500, 1,000, 2,000 μg/ml)로 동시 처리 시 농도 의존적으로 염증성사이토카인인 TNF-α와 IL-6의 단백질 생성을 통계적인 유의성(p<0.001)있게 억제함을 확인하였다. 또한 염증 유도된 RAW 264.7 세포에 PFE 동시 처리 시 NF-κB p65의 핵으로 이동이 차단됨과 iNOS와 COX-2의 단백질 발현을 감소시키는 것을 확인하였다. 이상의 연구결과를 통해 참콩 풍뎅이는 염증에 의해 활성화된 TLR-4 신호전달과정을 조절하는 NF-κB p65의 활성과 염증성사이토카인 TNF-α와 IL-6의 생성 및 염증성효소 iNOS와 COX-2의 생성을 억제하는 항염증 효능이 있음을 확인하였다.

• 생명과학회지
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• 제30권11호
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• pp.999-1006
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• 2020

최근 중추신경계에서 면역기능을 담당하는 미세아교세포(microglia)의 염증반응을 효율적으로 조절하는 것은 알츠하이머 병, 파킨슨 병 및 헌팅턴 병과 같은 퇴행성 뇌질환의 치료를 위한 중요한 타겟으로 인식되고 있다. 왕귀뚜라미(Teleogryllus emma)는 다양한 치료효능으로 인해 세계적으로 널리 이용되고 있으며, 본 연구팀에서는 최근 왕귀뚜라미의 전사체 분석을 통하여 항균활성을 가지는 다양한 종류의 새로운 항균 펩타이드(antimicrobial peptide; AMP) 후보들을 선별한 바 있다. 항균 펩타이드는 미생물에서부터 포유류까지 매우 다양한 종으로부터 발견되었으며, 현재는 항균활성뿐만 아니라 염증반응과 같은 다양한 질병의 치료제 개발을 위한 후보 물질로 관심을 받고 있다. 본 연구에서는 선행연구를 통하여 선별된 왕귀뚜라미 유래 항균 펩타이드들 중에서 Teleogryllusine(VKWKR-LNNNKVLQKIYFVKI-NH₂)으로 명명된 항균 펩타이드의 신경염증 억제 효능을 관찰하였다. Teleogryllusine의 신경염증 억제 효능을 관찰 하기 위하여 immortalized mouse microglia 세포주인 BV-2 세포에 Teleogryllusine을 1시간 전처리 한 후 LPS를 이용하여 BV-2 세포의 염증 반응을 유도하였다. 그 결과 Teleogryllusin은 최대 처리 농도인 80 ㎍/ml까지 세포독성 없이 nitric oxide (NO) 생성을 현저히 감소시킴을 확인할 수 있었다. 또한 염증반응 매개인자인 iNOS와 COX-2 및 cytokine (Il-6, TNF-α)의 발현을 유전자 수준과 단백질 수준에서 확인한 결과 Teleogryllusine 처리농도에 의존적으로 감소됨을 확인할 수 있었다. 또한 Teleogryllusine의 신경염증 억제작용 기작을 확인한 결과 mitogen activated protein kinases (MAPKs)와 IκB의 인산화 및 proteosome에 의한 IκB의 분해를 억제함으로서 BV-2 세포의 신경염증반응이 조절됨을 확인할 수 있었다. 이러한 결과로 보아 왕귀뚜라미 유래 Teleogryllusine 펩타이드는 신경염증반응에 의해 유도되는 퇴행성 뇌질환 치료 및 예방을 위한 의료용 소재로 사용될 수 있을 것으로 기대된다.

• 한국미생물·생명공학회지
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• 제44권3호
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• pp.236-245
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• 2016

밀배아유가 LPS로 자극된 대식세포에서 염증과 관련된 인자들에 미치는 영향을 살펴보기 위해, NO 및 전염증성 cytokine의 분비량과 iNOS 및 COX-2의 발현량, NF-κB p65와 MAPKs 활성화를 확인하였다. 또한 ICR 마우스를 이용하여 croton oil로 유도된 귀부종을 통해 귀두께 변화 및 귀조직을 관찰하였다. 그 결과, 밀배아유는 LPS로 인해 증가된 NO 및 전염증성 cytokine의 분비량을 감소시켰으며, 특히, IL-6와 TNF-α는 100 μg/ml 농도에서 90% 이상의 분비 억제효과를 나타내었다. LPS에 의해 증가된 iNOS와 COX-2의 발현은 100 μg/ml 농도로 밀배아유 처리시 발현 억제가 가장 크게 나타났으며, NF-κB p65의 발현도 밀배아유 농도가 증가함에 따라 발현 억제효과를 나타내었으며, 100 μg/ml 농도로 처리 시 억제 효과가 가장 컸다. 밀배아유는 MAPKs의 인산화를 억제시켰으며, 특히, ERK와 JNK의 인산화 억제가 농도 의존적으로 크게 효과를 나타내었다. 또한, 밀배아유는 croton oil로 유도된 귀부종에서 귀두께를 감소시켰으며, 경피와 진피의 두께 및 진피에 침윤된 비만세포의 수도 감소시킴을 확인하였다. 이상으로, 밀배아유는 NF-κB 및 MAPKs의 신호 경로 조절을 통한 염증성 매개 인자들의 발현 감소로 항염증 효과를 나타냄을 확인하여, 천연 항염증 소재로서 활용도가 가능함을 시사한다.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제36권5호
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• pp.392-401
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• 2010

Introduction: Diabetes mellitus, as a major health problem for the elderly has been shown to alter the properties of the bone and impair bone healing around a titanium implant in both humans and animals. The aim of this study was to examine the effect of adipose-derived stem cells on the healing process around a titanium implant in streptozotocin-induced diabetic rats. Materials and Methods: Thirteen rats were divided into two groups: adipose-derived stem cells injected group and a control group. A titanium screw implant (diameter: 2.0 mm, length: 3.5 mm) was placed into both tibia of 13 rats: 13 right tibia as the control group and 13 left tibia as the experimental group. The rats were sacrificed at different intervals (1, 2, and 4 weeks) after implantation for histopathology observations and immunohistochemistric analysis. Results: The histopathological findings revealed earlier new formed bone in the experimental group than the control group. In particular, at 1 week after implantation, the experimental group showed more newly formed bone and collagen around the implant than the control group. In immunohistochemistric analysis, osteoprotegerin (OPG) expression in the experimental group increased early compared to that of the control group until 2 weeks after implantation. However, after 2 weeks, OPG expression in the experimental group was similar to OPG expression in the control group. The receptor activator of nuclear factor ${\kappa}B$ ligand (RANKL) expression in the experimental group increased early compared to that of the control group, and then decreased at 2 weeks. After 2 weeks, the level of RANKL expression was similar in both groups. Conclusion: These results suggest that adipose-derived stem cells in implantation can promote bone healing around titanium, particularly in diabetes mellitus induced animals.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제36권5호
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• pp.386-391
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• 2010

Introduction: Bone resorption is a unique function of osteoclasts. Osteoclasts are a specialized macrophage polykaryon whose differentiation is regulated principally by macrophage colony-stimulating factors, receptor activator of nuclear factor ${\kappa}B$ ligand (RANK) ligand, osteoprotegerin (OPG), and interleukins (IL). Reflecting the integrin-mediated signals, osteoclasts develop a specialized cytoskeleton that allows it to establish an isolated micro-environment between itself and the bone, wherein matrix degradation occurs by a process involving proton transport. The levels of IL-1, IL-6, OPG, and prostaglandin $E_2$ ($PGE_2$) expression were evaluated to study the correlations between dental implant teeth and the adjacent teeth. Materials and Methods: The exudate of the gingival crevice acquired from dental implants, adjacent teeth, opposite teeth and contralateral teeth of 24 patients. Results: 1. The levels of IL-1, IL-6, OPG and $PGE_2$ expression in dental implant teeth were higher than those of the contralateral teeth. 2. IL-1 revealed a higher expression level in the adjacent teeth than in dental implant teeth. 3. The dental implant teeth and adjacent teeth did not show a remarkable difference in the level of IL-1 expression. 4. All the other cytokines were strongly expressed in the dental implant compared to the adjacent teeth. Conclusion: These results suggest that there might be close correlation between dental implant teeth and adjacent teeth in terms of the expressions of cytokines that affect the development and regulation of osteoclasts.

• 한국식품영양과학회지
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• 제39권1호
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• pp.64-70
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• 2010

본 연구는 MC3T3-E1 조골 전구세포에 대두추출물을 처리 후 그 조건배양액에서 파골세포분화 관련 국소인자 중파골세포분화 촉진인자(IL-$\beta$, IL-6, RANKL, TNF-$\alpha$, M-CSF) 및 파골세포로의 분화억제에 관여하는 국소인자인 OPG의 발현변화를 조골세포 조건배양액에서 살펴보았으며, 이 조골세포 조건배양액을 RAW264.7 파골전구세포에 처리 시 파골세포로의 분화를 억제정도를 TRAP 염색 및 관련 분화 지표의 발현을 통해 알아보았다. 대두추출물 처리한 조골세포 조건배양액에서 OPG의 발현이 농도 의존적으로 현저히 증가하였다. 그러나 강력한 파골세포 분화촉진인자로 알려진 IL-1$\beta$의 발현 역시 고농도의 대두추출물 처리 시 현저히 증가하여 고농도의 대두추출물을 처리한 CM 처리 시 TRAP 염색된 일부 파골세포를 확인하였고, 파골세포분화 관련지표인 MMP-9의 발현 또한 저농도 대두 추출물을 처리한 CM 처리에 비해 증가하였다. 이는 저농도의 대두추출물이 고농도의 대두추출물에 비해 파골세포분화억제에 효과적임을 의미하나, 추후 파골세포수준에서 분화억제 관련 기전연구가 필요할 것으로 생각되어진다.

• Tuberculosis and Respiratory Diseases
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• 제78권3호
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• pp.218-226
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• 2015

Background: Eph receptors and ephrin ligands have several functions including angiogenesis, cell migration, axon guidance, fluid homeostasis, oncogenesis, inflammation and injury repair. The EphA2 receptor potentially mediates the regulation of vascular permeability and inflammation in response to lung injury. Methods: Mice were divided into 3 experimental groups to study the role of EphA2 signaling in the lipopolysaccharide (LPS)-induced lung injury model i.e., IgG+phosphate-buffered saline (PBS) group (IgG instillation before PBS exposure), IgG+LPS group (IgG instillation before LPS exposure) and EphA2 monoclonal antibody (mAb)+LPS group (EphA2 mAb pretreatment before LPS exposure). Results: EphA2 and ephrinA1 were upregulated in LPS-induced lung injury. The lung injury score of the EphA2 mAb+LPS group was lower than that of the IgG+LPS group ($4.30{\pm}2.93$ vs. $11.45{\pm}1.20$, respectively; p=0.004). Cell counts (EphA2 mAb+LPS: $11.33{\times}10^4{\pm}8.84{\times}10^4$ vs. IgG+LPS: $208.0{\times}10^4{\pm}122.6{\times}10^4$; p=0.018) and total protein concentrations (EphA2 mAb+LPS: $0.52{\pm}0.41mg/mL$ vs. IgG+LPS: $1.38{\pm}1.08mg/mL$; p=0.192) were decreased in EphA2 mAb+LPS group, as compared to the IgG+LPS group. In addition, EphA2 antagonism reduced the expression of phospho-p85, phosphoinositide 3-kinase $110{\gamma}$, phospho-Akt, nuclear factor ${\kappa}B$, and proinflammatory cytokines. Conclusion: This results of the study indicated a role for EphA2-ephrinA1 signaling in the pathogenesis of LPS-induced lung injury. Furthermore, EphA2 antagonism inhibits the phosphoinositide 3-kinase-Akt pathway and attenuates inflammation.

• International Journal of Oral Biology
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• 제30권1호
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• pp.9-15
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• 2005

Recently, we reported that high extracellular calcium increased receptor activator of nuclear factor-${\kappa}B$ ligand (RANKL) expression via p44/42 mitogen-activated protein kinase (p44/42 MAPK) activation in mouse osteoblasts. However, the mechanism for p44/42 MAPK activation by high extracellular calcium is unclear. In this study, we examined the role of intracellular calcium increase in high extracellular calcium-induced RANKL induction and p44/42 MAPK activation. Primary cultured mouse calvarial osteoblasts were used. RANKL expression was highly induced by 10 mM calcium treatment. Ionomycin, a calcium ionophore, also increased RANKL expression and activated p44/42 MAPK. U0126, an inhibitor of MEK1/2, an upstream activator of p44/42 MAPK, blocked the RANKL induction by both high extracellular calcium and ionomycin. High extracellular calcium increased the phosphorylation of proline-rich tyrosine kinase 2 (Pyk2), one of the known upstream regulators of p44/42 MAPK activation. Bisindolylmaleimide, an inhibitor of protein kinase C, did not block RANKL induction and p44/42 MAPK activation induced by high extracellular calcium. 2-Aminoethoxydiphenyl borate, an inhibitor of inositol 1,4,5-trisphosphate (IP3) receptor, blocked the RANKL induction by high extracellular calcium. It also partially suppressed the activation of Pyk2 and p44/42 MAPK. Cyclosporin A, an inhibitor of calcineurin, also inhibited high calcium-induced RANKL expression in dose dependent manner. However, cyclosporin A did not affect the activation of Pyk2 and p44/42 MAPK by high extracellular calcium treatment. These results suggest that 1) the increase in intracellular calcium via IP3-mediated calcium release is necessary for RANKL induction by high extracellular calcium treatment, 2) Pyk2 activation, but not protein kinase C, following the increase in intracellular calcium might be involved in p44/42 MAPK activation, and 3) calcineurin-NFAT activation by the increase in intracellular calcium is involved in RANKL induction by high extracellular calcium treatment.

• 동의생리병리학회지
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• 제26권6호
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• pp.869-873
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• 2012

Balance between bone-forming osteoblasts and bone-resorbing osteoclasts is important in bone homeostasis. Unusual balance between bone-forming osteoblasts and bone-resorbing osteoclasts leads to bone diseases, such as osteoporosis. Saururus chinensis has been widely used in oriental medicine. Saururus chinensis has been known that has antioxidant and anticancer effect. But, the effect of Saururus chinensis in osteoclast differentation remains unknown. We examined the effect of Saururus chinensis in receptor activator of nuclear factor-${\kappa}B$ ligand (RANKL)-induced osteoclast differentiation. From the results of our study, we found that saururus chinensis clearly inhibited RANKL-induced osteoclast differentiation in bone marrow macrophages (BMM) in a dose dependent manner without toxicity. Saururus chinensis inhibited the phosphorylation of JNK, P38, AKT, and ERK induced by RANKL. The mRNA expression of NFATc1, TRAP, and OSCAR induced by RANKL was inhibited by Saururus chinensis treatment. Moreover Saururus chinensis suppressed the protein expression of c-Fos and NFATc1 in BMMs treated with RANKL. These results suggest that Saururus chinensis may be a useful drug in the treatment of bone-related disease.

• BMB Reports
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• 제49권10호
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• pp.554-559
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• 2016

Mycobacterium abscessus, a member of the group of non-tuberculous mycobacteria, has been identified as an emerging pulmonary pathogen in humans. However, little is known about the protective immune response of antigen-presenting cells, such as dendritic cells (DCs), which guard against M. abscessus infection. The M. abscessus gene MAB1843 encodes ᴅ-alanyl-ᴅ-alanine dipeptidase, which catalyzes the hydrolysis of ᴅ-alanyl-ᴅ-alanine dipeptide. We investigated whether MAB1843 is able to interact with DCs to enhance the effectiveness of the host's immune response. MAB1843 was found to induce DC maturation via toll-like receptor 4 and its downstream signaling pathways, such as the mitogen-activated protein kinase and nuclear factor kappa B pathways. In addition, MAB1843-treated DCs stimulated the proliferation of T cells and promoted Th1 polarization. Our results indicate that MAB1843 could potentially regulate the immune response to M. abscessus, making it important in the development of an effective vaccine against this mycobacterium.