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      • KCI등재

        파골세포 분화에 복령 추출물이 미치는 영향

        천윤희(Yoon Hee Cheon), 곽성철(Seong Cheoul Kwack), 오재민(Jaemin Oh), 최민규(Min-Kyu Choi), 김정중(Jeong Joong Kim), 곽한복(Han Bok Kwak), 이명수(Myeung Su Lee), 전병훈(Byung Hoon Jeon), 문서영(Seo Young Moon) 한의병리학회 2012 동의생리병리학회지 Vol.26 No.3

        Osteoporosis is an important public health issue in postmenopausal women. It is a major public health concern and is widely believed that osteoporosis results from imbalance between bone resorption and bone formation. Recently natural products from plants have been extensively studied as therapeutic drugs to treat and prevent various diseases. Hoelen (scientific name, Poria cocos) is a mushroom that is used in traditional Chinese medicine. Hoelen exhibits anti-inflammatory activity and has a protective effect on tumor progression. However, the effect of hoelen in osteoclast differentiation remains unknown. Thus, we examined the effect of hoelen in receptor activator of nuclear factor-κB ligand (RANKL)-induced osteoclast differentiation. Hoelen significantly inhibited RANKL-induced osteoclast differentiation in bone marrow-derived macrophages (BMMs) in dose dependent manner without toxicity. Also, we showed that hoelen significantly inhibited the mRNA expression of tartrate-resistant acid phophatase (TRAP) and nuclear factor of activated T cells 1 (NFATc1) in BMMs treated with RANKL. In Particular, hoelen greatly inhibited the protein expression of NFATc1. Ectopic expression of NFATc1 partially reverses hoelen-mediated inhibition of osteoclast differentiation. Taken together, our results demonstrated that hoelen may be useful treatment option of bone-related disease such as osteoporosis, reumatoid arthritis, and periodontitis.

      • KCI등재

        대추 물 추출물이 RANKL에 의해 유도되는 파골세포 분화에 미치는 영향

        윤강휴(Yoon Kang Hugh), 백종민(Jong Min Baek), 김주영(Ju Young Kim), 곽성철(Seong Cheoul Kwak), 천윤희(Yoon Hee Cheon), 전병훈(Byung Hoon Jeon), 이창훈(Chang Hoon Lee), 최민규(Min Kyu Choi), 오재민(Jaemin Oh), 이명수(Myeung Su Lee), 김정중(Jeo) 한의병리학회 2014 동의생리병리학회지 Vol.28 No.1

        Bone homeostasis is maintained by balance between bone resorbing-osteoclasts and bone forming-osteoblasts. Excessive osteoclastic bone resorption plays a critical role in bone destruction in pathological bone diseases such as osteoporosis, rheumatoid arthritis, and periodontal disease. Many compounds derived from natural products have pharmacological applications and have therapeutic value for treating or preventing several bone diseases characterized by excessive bone resorption. To discover new compounds that can act as anti-resorptive agents, we screened for natural compounds that regulate osteclast differentiation, and found that water extract of Ziziphus Jujuba Mill (WEZJ) has inhibitory effects on osteoclast differentiation. In this study, WEZJ clearly inhibits the osteoclast differentiation in the presence of receptor activator of nuclear factor kB (RANKL), macrophage colony-stimulating factor (M-CSF) without cytoxicity by blocking activation of nuclear factor of activated T cells (NFAT)c1, and c-Fos. In signaling pathway, the phosphorylation of Akt, p38, c-Jun N-terminal kinases (JNK), extracellular signal-regulated kinases (ERK) and the expression of osteoclast-associated receptor (OSCAR), tartrate-resistant acid phosphates (TRAP), Integrin av, Integrin b3, Cathepsin K are suppressed, too. These result suggest that WEZJ may have therapeutic value for treating or preventing several bone diseases characterized by excessive bone destruction.

      • KCI등재

        Baicalein이 골수유래대식세포의 파골세포 분화에 미치는 효과

        윤지광(Ji Kwang Yun), 천윤희(Yoon-Hee Cheon), 김주영(Ju-Young Kim), 곽성철(Seong Cheoul Kwak), 윤강휴(Kang Hue Yoon), 백종민(Jong Min Baek), 이명수(Myeong Su Lee), 오재민(Jaemin Oh), 박종태(Jongtae Park) 대한체질인류학회 2014 대한체질인류학회지 Vol.27 No.2

        현대사회가 급속한 노령화 시대에 접어들면서 뼈 건강에 대한 중요성이 증가됨으로써 최근, 천연물을 이용한 골질환의 치료제를 개발하기 위한 연구가 활발히 이루어지고 있다. Baicalein은 기존에 항암, 항염증, 항산화의 효과가 있고, MC3T3-E1 조골세포주의 분화를 촉진하며, RAW264.7세포주의 분화는 억제한다는 보고가 있다. 그러나 골수유래대식세포를 이용한 파골세포 분화의 억제기전에 대해서는 정확히 알려진 바가 없다. 그리하여 본 연구에서는 골수유래대식세포의 파골세포 분화에 있어 baicalein의 억제기전을 밝히고, 골수기질세포의 조골세포 분화에 미치는 영향을 탐색하기 위한 실험을 진행하였다. 파골세포 분화를 위해 골수유래대식세포를 사용하여 분화, mRNA 및 단백질 발현을 확인하였다. 조골세포 분화를 위해 골수기질세포를 사용하여 분화, ALP활성 및 미네랄 침착능을 확인하였다. Baicalein은 골수유래대식세포에서 RANKL로 유도한 파골세포로의 분화를 세포독성 없이 농도 의존적으로 억제하였다. 억제기전으로는 Akt, JNK, PLCγ2의 인산화를 억제하였고, NF-κB의 활성화를 억제하여 파골세포 분화의 필수 유전자인 c-Fos 및 NFATc1의 발현 역시 억제하였다. 또한 TRAP, OSCAR, cathepsin K와 DCSTAMP의 mRNA 수준의 발현을 감소시켰다. 이와 더불어, baicalein이 골 형성의 기능을 갖는 조골세포의 분화를 촉진하는 효과를 골수기질세포에서 확인하였다. Baicalein은 골수유래대식세포에서 파골세포 분화를 억제하고, 동시에 골수기질세포를 이용한 조골세포 분화를 높이는 양면적인 효과를 가지는 물질로, 향후 골 소실 질환 치료제 개발에 중요 후보물질이 될 것으로 사료된다. As prediction of rapidly aging society, bone health is considered increasingly important and received more attention than ever. Bone health is regulated by balancing between bone resorptive osteoclasts and bone formative osteoblasts. Disruption of balance between bone-resorbing osteoclasts and bone-forming osteoblasts results in bone disease. Natural products have recently received much attention as an alternative tool for the development of novel therapeutic strategy. Baicalein is reported it has anti-cancer, anti-inflammatory and antioxidant effects. Baicalein also has been known that it has both promotive effect on MC3T3-E1 cell line and inhibitory effect on RAW 264.7 cell line. However, the inhibitory mechanism of baicalein using bone marrow derived macrophages (BMMs) on osteoclast differentiation remains not clear. In this study, the suppressive mechanism by baicalein on osteoblast differentiation was evaluated. Bicalein inhibited receptor activator of nuclear factor-κB ligand (RANKL)-induced osteoclast differentiation in BMMs in a dose dependent manner without any toxicity. Baicalein suppressed phosphorylation of protein kinaseB (Akt), c-Jun N-terminal kinases (JNK) and phosphoinositide-specific phospholipaseCγ2 (PLCγ2). Furthermore, Baicalein suppressed the induction of RANKL-induced c-Fos and Nuclear factor of activated T cell c1 (NFATc1), essential genes on osteoclastogenesis. In BMMs, Bicalein inhibited the mRNA expression of tartrate-resistant acid phosphatase (TRAP), osteoclast-associated receptor (OSCAR), cathepsinK, dendritic cell-specific transmembrane protein (DCSTAMP). Moreover, baicalein promoted differentiation of osteoblast on bone marrow stromal cells (BMSCs). Taken together, these results suggest that baicalein has a potential for treating bone lytic diseases, such as osteoporosis, periodontitis, and rheumatoid arthritis.

      • KCI등재

        파골세포의 분화와 뼈 흡수에 미치는 칡(Pueraria lobata)의 영향

        백종민(Jong Min Baek), 윤강휴(Kang Hugh Yoon), 안성준(Sung-Jun Ahn), 박선향(Sun-Hyang Park), 천윤희(Yoon-Hee Cheon), 김주영(Ju-Young Kim), 오재민(Jaemin Oh) 대한체질인류학회 2014 대한체질인류학회지 Vol.27 No.4

        칡은 뼈소실 모델인 난소를 적출한 마우스와 랫드에서 뼈형성을 촉진하다고 밝혀진 바 있다. 그러나 뼈파괴세포의 분화와 뼈흡수능에 미치는 칡의 효과와 기전은 아직 밝혀지지 않았다. 그러므로 본 연구에서는 칡이 recetor activator of nuclear factor-κB ligand (RANKL)과 macrophage colony stimulating factor (M-CSF)로 유도한 뼈파괴세포의 분화에 미치는 효과와 그 기전을 연구하였다. 우선적으로 뼈파괴세포 분화에 억제효과를 확인하기 위하여 마우스에서 분리한 큰포식세포에 칡을 농도별로 처리하여 tartrate-resistant acid phosphatase (TRAP) staining을 시행하였다. 또한 칡이 작용하는 기전을 알아보기 위하여 western blot 분석과 RT-PCR을 시행하였고, 성숙한 뼈파괴세포의 뼈흡수능에 미치는 영향을 알아보고자 hydroxyapatite 플레이트로 실험을 진행하였다. 그 결과, 칡은 뼈파괴세포 분화단계의 필수적인 초기 신호전달경로인 p38, Akt, c-Jun N-terminal kinase (JNK), IκB의 인산화를 억제하였다. 또한 nuclear factor of activated T cells (NFAT)c1과 c-Fos의 비활성화를 유도함으로써 뼈파괴세포와 관련된 유전자인 osteoclast-associated receptor (OSCAR), TRAP, Integrin β3, osteoclast stimulatory transmembrane protein (OC-STAMP), dendritic cell-specific transmem-brane protein (DC-STAMP)의 발현을 저해하였다. 특히, hydroxyapatite로 코팅된 플레이트를 통해서 뼈흡수능을 억제하는 칡의 효과를 확인하였으며, 이와 깊은 연관성을 가지는 유전자인 Cathepsin K의 발현 또한 저해하였다. 이러한 결과를 통하여 뼈파괴세포 분화와 뼈흡수능을 저해하는 칡의 효과와 이와 관련된 분자적 기전에 대하여 규명하였다. Previous researches have proved that Pueraria lobata up-regulates bone mineral contents and bone mineral density in bone-loss model, ovariectomized mice and orchidectomized rats. However, the precise effects and mechanisms of Pueraria lobata on osteoclast differentiation and bone resorbing activity of mature osteoclasts still remains unknown. Therefore, we investigated the effect and mechanism of Pueraria lobata on receptor activator of nuclear factor-κB ligand (RANKL) and macrophage colony stimulation factor (M-CSF)-induced osteoclast differentiation in bone marrow macro-phages (BMMs). First of all, we treated BMMs derived from mice with various concentrations of Pueraria lobata in order to perform screening by tartrate-resistant acid phosphatase (TRAP) staining. Also, we conducted western blotting and RT-PCR for the purpose of verifying the treatment mechanism of Pueraria lobata and lastly, we used hydroxyapatite-coated plate to evaluate the effects of Pueraria lobata on bone resorbing activity of mature osteoclasts. As a result, Pueraria lobata has inhibitory effect on phosphorylation of p38, Akt, c-Jun N-terminal kinase (JNK), and IκB which are essential early signaling pathway of osteoclastogenesis. Also, the inactivation of nuclear factor of activated T cells (NFAT)c1, and c-Fos which is caused by Pueraria lobata is followed by the suppression effects of Pueraria lobata on osteoclastrelated various genes, osteoclast-associated receptor (OSCAR), TRAP, Integrin β3, osteoclast stimulatory transmembrane protein (OC-STAMP), and dendritic cell-specific transmembrane protein (DC-STAMP). Particularly, Pueraria lobata blocks the formation of pit area on hydroxyapatite-coated plate in a dose-dependent manner as well as the mRNA expression of Cathepsin K, which is associated with bone resorbing activity. These results demonstrate the molecular mechanism relating to anti-osteoclastogenesis effect of Pueraria lobata as well as the inhibitory effect of Pueraria lobata on mature osteoclast formation and bone resorbing activity.

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