• 동의생리병리학회지
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• 제25권4호
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• pp.669-673
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• 2011

To prevent and treat the osteoporotic fracture, more attention should be paid in old age patients. Osteoclast which has ability to bone resorption is originated from hematopoietic cell line and plays a key role osteoporotic bone loss. Rubi Fructus has been widely used in Oriental medicine. Extracts of the leaves and fruit of Rubus species have been used in various countries as natural remedies to treat diabetes, infections, colic, and burns. However, the effect of extract of Rubi Fructus (fruit of Rubus coreanus Miq.) in osteoclast differentiation remains unknown. Thus, we evaluated the effect of Rubi Fructus on receptor activator of nuclear factor-kB ligand (RANKL)-induced osteoclast differentiation. Here we found that Rubi Fructus significantly inhibited osteoclast differentiation induced by RANKL. Rubi Fructus suppressed the activation of p38 pathway and NFkB in bone marrow macrophages (BMMs) treated with RANKL. Also, Rubi Fructus significantly inhibited the mRNA expression of c-Fos, tartrate-resistant acid phosphatase (TRAP), osteoclast-associated receptor (OSCAR), nuclear factor of activated T cells (NFAT)c1 and cathepsin K in BMMs treated with RANKL. Particularly, Rubi Fructus greatly inhibited the protein expression of c-fos and NFATc1. especially in the case of NFATc1 expression, a master transcription factor of the differentiation of osteoclasts is very important step for osteoclastogenesis. Taken together, our results demonstrated that Rubi Fructus may be useful treatment option of bone-related disease such as osteoporosis and rheumatoid arthritis.

• The Korean Journal of Physiology and Pharmacology
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• 제16권1호
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• pp.31-36
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• 2012

The receptor activator of NF-${\kappa}B$ ligand (RANKL) signal is an activator of tumor necrosis factor receptor-associated factor 6 (TRAF6), which leads to the activation of NF-${\kappa}B$ and other signal transduction pathways essential for osteoclastogenesis, such as $Ca^{2+}$ signaling. However, the intracellular levels of inositol 1,4,5-trisphosphate ($IP_3$) and $IP_3$-mediated cellular function of RANKL during osteoclastogenesis are not known. In the present study, we determined the levels of $IP_3$ and evaluated $IP_3$-mediated osteoclast differentiation and osteoclast activity by RANKL treatment of mouse leukemic macrophage cells (RAW 264.7) and mouse bone marrow-derived monocyte/macrophage precursor cells (BMMs). During osteoclastogenesis, the expression levels of $Ca^{2+}$ signaling proteins such as $IP_3$ receptors ($IP_3Rs$), plasma membrane $Ca^{2+}$ ATPase, and sarco/endoplasmic reticulum $Ca^{2+}$ ATPase type2 did not change by RANKL treatment for up to 6 days in both cell types. At 24 h after RANKL treatment, a higher steady-state level of $IP_3$ was observed in RAW264.7 cells transfected with green fluorescent protein (GFP)-tagged pleckstrin homology (PH) domains of phospholipase C (PLC) ${\delta}$, a probe specifically detecting intracellular $IP_3$ levels. In BMMs, the inhibition of PLC with U73122 [a specific inhibitor of phospholipase C (PLC)[ and of $IP_3Rs$ with 2-aminoethoxydiphenyl borate (2APB; a non-specific inhibitor of $IP_3Rs$) inhibited the generation of RANKL-induced multinucleated cells and decreased the bone-resorption rate in dentin slice, respectively. These results suggest that intracellular $IP_3$ levels and the $IP_3$-mediated signaling pathway play an important role in RANKL-induced osteoclastogenesis.

• Molecules and Cells
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• 제43권4호
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• pp.340-349
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• 2020

Oleoylethanolamide (OEA), a bioactive lipid in bone, is known as an endogenous ligand for G protein-coupled receptor 119 (GPR119). Here, we explored the effects of OEA on osteoclast differentiation, function, and survival. While OEA inhibits osteoclast resorptive function by disrupting actin cytoskeleton, it does not affect receptor activator of nuclear factor-κB ligand (RANKL)-induced osteoclast differentiation. OEA attenuates osteoclast spreading, blocks actin ring formation, and eventually impairs bone resorption. Mechanistically, OEA inhibits Rac activation in response to macrophage colony-stimulating factor (M-CSF), but not RANKL. Furthermore, the OEA-mediated cytoskeletal disorganization is abrogated by GPR119 knockdown using small hairpin RNA (shRNA), indicating that GPR119 is pivotal for osteoclast cytoskeletal organization. In addition, OEA induces apoptosis in both control and GPR119 shRNA-transduced osteoclasts, suggesting that GPR119 is not required for osteoclast apoptosis. Collectively, our findings reveal that OEA has inhibitory effects on osteoclast function and survival of mature osteoclasts via GPR119-dependent and GPR119-independent pathways, respectively.

• Journal of Periodontal and Implant Science
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• 제47권6호
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• pp.381-387
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• 2017

Purpose: The aim of this study was to evaluate volumetric and histologic changes in edentulous alveolar ridge areas after ridge preservation using basic fibroblast growth factor-2 (bFGF-2) in combination with collagenated biphasic calcium phosphate (BCP). Methods: The experiments were performed in 6 adult male beagle dogs. The following 3 groups were created: 1) ridge preservation with bFGF-2 and collagenated BCP (experimental group), 2) ridge preservation with collagenated BCP (positive control group), and 3) a negative control group in which no ridge preservation procedure was performed. Volumetric change analysis was performed using an optical scanner and casts. Histological observations were made using light microscopy. Results: After the initial swelling subsided, the magnitude of the volumetric change in the experimental group and positive control group was smaller than in the negative control group. In the experimental group, a distinct trend was observed for the resorption of residual bone and collagen fibers at 4 weeks and for more mature bone and faster healing at 12 weeks. Conclusions: Based on the findings of the present study, bFGF-2 may be considered for use as a therapeutic molecule in ridge preservation procedures.

• 대한한방부인과학회지
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• 제25권1호
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• pp.1-10
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• 2012

Purpose: This study was conducted to evaluate the inhibitory effect of Achyranthis Radix extract(ARE) loaded hydrogel on osteoclast differentiation. Methods: MTT-assay was performed to estimate cytotoxicity of ARE, Achyranthis Radix-alginate hydrogel disk(ARHD) in bone marrow macrophages stimulated(BMMs) with human receptor activator of nuclear factor-${\kappa}B$ ligand(RANKL), human macrophage-colony stimulating factor(M-CSF). Tartrate resistant acid phosphatase staining and RT-PCR were performed to know the inhibitory effect on osteoclast differentiation. Reactive oxygen species and actin ring formation were analysed to observe the effect of ARHD. Results: ARE has no cytotoxicity at the concentration of 0.1 $mg/m{\ell}$ or lower. ARE decreased the number of TRAP positive cells in RANKL, M-CSF stimulated BMMs and the gene expression. ARHD has no cytotoxicity at the concentration of 10 ${\mu}g/m{\ell}$ (24, 48hours), 50 ${\mu}g/m{\ell}$ (24 hours). ARHD restrained the synthesis of reactive oxygen species and the formation of actin ring. Conclusions: Achyranthis Radix has the inhibitory effect of osteoclast differentiation and bone resorption. Further studies are needed to treat osteoporosis by Achyranthis Radix.

• 동의생리병리학회지
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• 제26권2호
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• pp.160-165
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• 2012

Osteoporotic fracture became a serious social problem, which related with mortality and morbidity in old age population. Osteoclast which is responsible for bone resorption is originated from hematopoietic cell line and plays a key role osteoporotic bone loss. Cynanchum wilfordii (Asclepiadaceae) roots have been used in Korean folk medicine for the treatment of diabetes mellitus and aging progression. Also, recent studies have shown that the extract and fractions of Cynanchi Wilfordii Radix have various pharmacological actions including scavenging free radicals, enhancing immunity, reducing high serum cholesterol, and anti-tumor activity. However, the effect of extract of Cynanchi Wilfordii Radix in osteoclast differentiation had not been reported. Thus, we evaluated the effect of Cynanchi Wilfordii Radix on receptor activator of nuclear factor-${\kappa}B$ ligand (RANKL)-induced osteoclast differentiation. Through our study, we found that Cynanchi Wilfordii Radix significantly inhibited osteoclast differentiation induced by RANKL. Cynanchi Wilfordii Radix suppressed the activation of p38 pathway and $NF{\kappa}B$ in bone marrow macrophages (BMMs) treated with RANKL. Also, Cynanchi Wilfordii Radix significantly inhibited the mRNA expression of c-Fos, tartrate-resistant acid phosphatase (TRAP), osteoclast-associated receptor (OSCAR), nuclear factor of activated T cells (NFAT)c1 and cathepsin K in BMMs treated with RANKL. Particularly, Cynanchi Wilfordii Radix inhibited the protein expression of c-fos and NFATc1. Taken together, our results demonstrated that Cynanchi Wilfordii Radix may be useful treatment option of bone-related disease such as osteoporosis leads to fracture of bone and rheumatoid arthritis.

• Journal of Acupuncture Research
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• 제32권3호
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• pp.69-81
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• 2015

Purpose : The first purpose of this study is to find out whether the water extract of Rehmanniae Radix Preparat(RRP), Cuscutae Semen(CS) and their combination(Ssangbohwan, SBH) have the effect of suppressing Receptor activator of nuclear factor kappa-B ligand(RANKL)-induced osteoclast differentiation. The second purpose of this study is to find out whether the water extract of RRP, CS and SBH have the effect of inhibiting osteoporosis in an osteoporosis model induced by lipopolysaccharide(LPS). Methods : After promoting differentiation of osteoclasts by treating the RANKL, we observed the effect by the administration of RRP, CS and SBH. In addition, by means of Reverse transcription polymerase chain reaction(RT-PCR), we assayed mRNA expression levels of NFATc1, c-Fos, TRAP and GAPDHS(Glyceraldehyde-3-phosphate dehydrogenase, spermatogeni) from bone marrow macrophages(BMMs). Similarly, the protein expression levels of NFATc1 (Nuclear factor of activated T-cells, cytoplasmic1), C-Fos, MAPKs(Mitogen-activated protein kinases) and ${\beta}$-actin in cell lysates were analyzed by means of Western Blotting. Finally, we determined the anti-osteoporotic effects of RRP, CS and SBH, through the use of Lipopolysaccharide-induced bone-loss mouse. Results : RRP, CS and SBH showed remarkable inhibitive effect on RANKL-treated osteoclast differentiation without cytotoxicity. SBH inhibited the phosphorylation of p38, Jun N-terminal kinases(JNK), and I-${\kappa}B$ and down-regulated the induction of c-Fos and NFATc1 by RANKL. RRP, CS suppressed degradation of I-${\kappa}B$, but it did not affect c-Fos and NFATc1 by RANKL. Lastly, in vivo data showed that RRP and SBH prevented bone erosion by LPS treatment. Conclusions : These results demonstrate SBH can be effective remedy for bone-loss diseases such as osteoporosis.

• Journal of Oral Medicine and Pain
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• 제31권1호
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• pp.17-25
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• 2006

혈관신생(angiogenesis)은 골조직을 포함하는 모든 조직의 발생 및 개조(remodeling) 과정에 필요하다. 본 연구는 혈관 신생에 관여하는 단백질인 angiopoietin-2가 골대사에서 미치는 영향을 알아보기 위하여 세포수준에서 관찰하였다. 즉 조골 세포에 미치는 영향을 알아보기위하여 세포생존률, 염기성인산분해효소 활성, gelatinase 활성 및 nitric oxide 생성을 관찰하였고, 파골세포에 미치는 영향을 알아보기 위하여 tartrate-저항성 인산분해효소 양성인 다핵세포의 형성 및 파골세포전구세포 배양 후 흡수면적을 측정함으로써 관찰하였다. Angiopoietin-2는 조골세포의 세포생존률 및 염기성 인산분해효소 활성을 증가시켰으며, gelatinase와 nitric oxide 생성의 증가시켰다. 또한 angiopoietin-2는 파골세포 생성 및 활성을 감소시켰다. 따라서 angiopoietin-2는 골수의 미세환경에서 세포의 조절작용을 하는 단백질로 여겨진다.

• Pediatric Gastroenterology, Hepatology & Nutrition
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• 제9권2호
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• pp.226-232
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• 2006

목 적: 체중과 지방량이 골밀도와 밀접한 연관이 있는 것으로 알려져 왔다. 비만아에서는 지방 조직에서 합성 분비되는 leptin이 증가되는데, 증가된 leptin이 비만아의 골 대사에 미치는 영향에 대해 알아보고자 연구를 하였다. 방 법: 2005년 11월부터 2006년 5월까지 조선대학교병원 외래를 방문한 50명의 남자(비만아 25명, 정상 체중의 대조군 25명)를 대상으로 하였다. 체질량 지수, 체지방률, 삼두박근 피부주름 두께와 견갑골하 피부주름 두께를 측정하고 혈청 leptin과 골대사 지표인 B-ALP, CICP 및 total DPD 농도를 검사하였다. 비만군과 대조군의 신체 계측, leptin 및 골대사 지표 농도를 비교하고, leptin과 골대사 지표와의 상관성을 알아보았다. 결 과: 비만군에서 체질량 지수(r=0.551, p<0.05), 체지방률(r=0.584, p<0.05)과 유의한 양의 상관관계가 있었다. Leptin은 골형성 지표인 B-ALP (r=-0.613, p=0.026) 및 CICP (r=-0.583, p=0.037)와 유의한 음의 상관성을 보였다. 그러나 골흡수 지표인 total DPD (r=0.103)와는 유의한 상관성이 없었다. 대조군에서 leptin과 유의한 상관성이 있는 인자는 없었다. 골대사에 영향을 주는 체중을 통제하고 상관성을 알아본 결과에서도 비만군에서 leptin은 B-ALP (r=-0.728, p=0.007) 및 CICP (r=-0.684, p=0.014)와 유의한 음의 상관성을 보였으나 total DPD와는 연관성이 없었다. 대조군에서는 leptin과 연관성 있는 골대사 지표는 없었다. 비만군에서 골대사 지표에 영향을 미치는 인자에 대해 다중 단계별 회귀 분석을 시행한 결과 종속변수인 B-ALP는 leptin과 유의한 음의 상관성을 보였으며(Y=-39.653X+356.341, p=0.026), CICP도 leptin과 유의한 음의 상관성을 보였다(Y=-13.437X+116.013, p=0.037). 결 론: 비만아에서 leptin은 골형성 지표인 B-ALP 및 CICP와 음의 상관성이 있었으며, 골 흡수가 아닌 골 형성을 저하시키는 것을 알 수 있었다.

• 구강회복응용과학지
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• 제31권3호
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• pp.242-252
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• 2015

무치악 환자에서는 치아가 상실되면서 주위 치조골의 개조와 흡수가 일어나고, 이에 따라 총의치의 유지력 감소, 저작 효율의 저하와 통증으로 인해 의치 사용에 어려움을 겪게 된다. 이를 개선하기 위해 하악에 2 - 4개의 임플란트를 식립하고 어태치먼트에서 유지 또는 지지를 얻는 피개의치가 바람직한 치료 방법으로 고려되고 있다. 본 증례들은 하악 완전 무치악 환자들을 악간 관계, 골흡수 정도, 안모지지 등을 평가하여 하악에 2개의 임플란트 식립을 고려하고, 다양한 종류의 어태치먼트를 이용한 임플란트 피개의치 수복을 계획하여 치료하였다. 주기적인 경과 관찰 결과 심미적, 기능적으로 만족할만한 결과를 얻었기에 이를 보고하는 바이다.