• 약학회지
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• 제41권1호
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• pp.1-6
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• 1997

Rubus coreantim (Rosaceae), which has been used in oriental traditional medicine as the remedies for impotence, pollution and premature ejaculation, bet-wetting and frequ ency of urination. is a fruit drug cultivating throughout southern Korea. The leaves of Rubus coreanum were extracted with 80% acetone. The extracts were separated by Ambellite XAD-2, Sephadex LH-20. MCI-gel CHP 20P and Cosmosil 75 C$_{18}$ OPN. And three flavonoids were isolated. The structures of these compounds were elucidated as quercetin-3-O-${\beta}$-D-glueuronopyranoside(l). quercetin-3-O-${\beta}$-D-glueuronopyranosyl methyl ester(2), quercetin-3-O-${\beta}$-D-xylopyranosyl-(2${\rightarrow}$1)-${\beta}$-glucopyranoside(3) on the basis of physicochemical and spectroscopic evidences.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2020년도 춘계학술대회
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• pp.90-90
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• 2020

Panax ginseng C.A. Meyer (P. ginseng) is known to exert a wide range of pharmacological effects both in vitro and in vivo. Although studies on ginsenoside, antioxidant activity, and anticancer effect of wild simulated ginseng (WSG) have been conducted, there is little research on the effect of WSG on bone metabolism. In this study, we investigated the potential anti-osteoporotic properties of WSG on the growth and differentiation of MC3T3-E1 cells. WSG significantly increased the viability and proliferation of MC3T3-E1 cells. WSG activated intracellular alkaline phosphatase (ALP) activity in MC3T3-E1 cells. In addition, WSG increased the mineralized nodules in MC3T3-E1 cells. Furthermore, WSG increased the expression of genes such as Runx2, ALP, OPN and OCN associated with osteoblast growth and differentiation in a dose-dependent manner.

• 생약학회지
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• 제25권1호
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• pp.11-19
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• 1994

Gleditsia japonica var. koraiensis NAKAI(Leguminosae) is commonly distributed in Korea and has been used as a folk medicine in the treatment of bronchitis, neoplasm and blennorrhgia in the Orient. The aqueous acetone extract of the leaves of G. japonica was subjected to a combination of Sephadex LH-20, Cosmosil $75C_{18}-OPN$, TSK-gel Toyopearl HW 40F, Avicel cellulose, and MCI-gel CHP 20P chromatographies with various solvent systems. Twelve compounds were isolated and confirmed to be vitexin(1), isovitexin(2), orientin(3), isoorientin(4), 4-caffeoyl quinic acid(5), 5-caffeoyl quinic acid(6), 3, 5-dicaffeoyl quinic acid(7), 4, 5-dicaffeoyl quinic acid(8), caffeic acid(9), quercetin(10), isoquercitrin(11) and luteolin-7-O-glucoside(12), on the basis of chemical and spectroscopic evidences.

• Journal of Acupuncture Research
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• 제23권5호
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• pp.69-77
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• 2006

Background : Mesenchymal stem cells (MSCs) are present in most of the tissue matrix, taking part in their regeneration when injury or damage occurs. The aim of this study was to investigate the presence of cells with pluripotential characteristics in human subchondral bone and the capacity of these cells to differentiate to osteoblast. Methods : Human subchondral bone were digested with collagenase. Isolated cells were cultured with a-MEM, 15% FBS, 10-8M dexamethasone and 50 ng/mL ascoric acid. Cells from 0 day(isolated cells), 7 day (first subculture) and 14 days (third subculture) were used to carry out phenotypic characterization experiments flowcytometry analysis with 11 monoclonal antibodies) and osteogenic differentiation experiments. Osteogenic differentiation of cells was assessment by quantification of bone extracellular matrix components by following analysis: alkaline phosphatase(ALP) stains to detect ALP activity, RT-PCR and western blot to detect osteocalcin (OCN), osteopontin (OPN) and type I collagen(Col I), and Alizarin red stains to detect calcium deposition. Results : Flowcytometry analyses showed that in our population more than 98% of cells were positive for MSC markers: SH-2(CD105, 99%), CD29 (95%), CD73 (95%). Cells were negative for hematopoietic markers (CD11b, CD34, and CD45). Furthermore, cells showed positive stain to multipotent markers such as CDl17 (c-kit) (15.1%), and CD166 (74.9%), and cell adhesion molecules such as CD54 (78.1%) and CD106 (63.5%). The osteogenic specific marker analyses showed that the culture of these cells for 7 and 14 days stimulates ALP, OCN, OPN and Col I synthesis by RT-PCR and Western blot analysis. Also, after 14 days in the culture of MSCs induces mineralization by Arizarin red stain. Conclusion : In this work, we demonstrated a new and efficient method for osteoblastic differentiation of human subchondral bone stem cells. As MSCs takes part in reparative processes of adult tissues, these cells could play an important role in osteogenesis.

• 한국자원식물학회지
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• 제33권4호
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• pp.227-236
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• 2020

고령사회에서 노년기 건강의 큰 문제로 대두되고 있는 골다공증은 특히 폐경 후 여성들에게서 가장 그 발생빈도가 높게 나타났으며, 현재 골다공증 예방 및 치료에 사용되고 있는 약제는 대부분 골흡수 억제제로써 진행된 골소실을 회복 시킬 수는 없기 때문에 골형성 증가를 통한 골다공증 예방과 치료에 관한 연구가 활발히 이루어지고 있다. 산양삼(cultivated wild Panax ginseng, CWP)에 대한 연구는 다수가 원기회복, 자양강장 및 면역증강 효과 등에 대한 것이나 골대사에 미치는 영향에 대한 연구는 거의 없는 실정이다. 이에 본 연구에서는 산양삼 추출물이 조골세포에서 골관련 유전자 발현에 미치는 영향을 확인함으로써 골다공증 예방 및 치료 효과를 갖는 천연 소재로의 활용 가능성을 검토하고자 하였다. 산양삼 추출물 처리가 조골 세포의 증식에 미치는 영향을 알아보기 위해 MTT assay를 실시하였고, MC3T3-E1 세포생존률은 FBS가 첨가되지 않은 배양액만 처리한 대조군과 산양삼 추출물을 처리한 실험군 모두에서 동일한 수준으로 나타났으며 이로써 산양삼 추출물의 안전성을 확인할 수 있었다. 또한 산양삼 추출물을 처리한 실험군과 대조군과의 세포증식률을 비교하였을 때 산양삼 추출물 50 ㎍/mL 농도 처리군에서 유의적으로 세포증식이 촉진되었으며 25 ㎍/mL과 100 ㎍/mL 농도 처리군에서도 대조군보다 높은 경향을 나타내었다. 산양삼 추출물이 조골 세포의 활성에 미치는 영향을 알아보기 위해 조골세포의 분화초기 표지인자인 ALP활성을 측정하였으며 그 결과 모든 산양삼 추출물 처리군이 대조군과 비교하여 유의적으로 높은 활성을 나타내었으며 특히 산양삼 추출물 50 ㎍/mL 농도 처리군에서 가장 높은 활성을 나타내었다. 산양삼 추출물의 농도에 따른 석회화 형성도를 확인하기 위해 무기질화된 세포의 기질을 alizarin red로 염색하였고 산양삼 추출물을 처리한 실험군과 대조군과의 석회화 형성도를 비교하였을 때 산양삼 추출물 50 ㎍/mL 농도 처리군에서 유의적으로 석회화 형성이 촉진되었으며 25 ㎍/mL과 100 ㎍/mL 농도 처리군에서도 대조군보다 높은 경향을 나타내었다. 산양삼 추출물이 MC3T3-E1 조골세포에서 골 형성 관련 유전자 발현에 미치는 영향을 확인하기 위해 Runx2, ALP, OPN, OCN등의 유전자를 정량 real-time PCR을 통해 분석하였으며 대조군과 비교하여 모든 산양삼 추출물 처리군에서 농도 의존적이고 유의적으로 골 형성 관련 유전자발현이 증가되었다. 따라서 산양삼추출물이 골 형성 관련 유전자인 Runx2, ALP, OPN, OCN 발현을 증가시켜MC3T3-E1 조골세포의 분화를 촉진하고, 골 석회화 형성 촉진에 기여하였을 것으로 사료된다. 그러나 산양삼 추출물이 골형성과 관련하여 어떠한 기전으로 유전자의 발현을 조절하였는지에 대한 유전자 및 단백질 수준의 추가적인 연구와 산양삼 추출물의 분화 촉진과 석회화 형성능이 산양삼의 사포닌계 진세노사이드 성분의 영향인지에 대한 후속 연구가 필요할 것으로 사료된다.

• BMB Reports
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• 제49권3호
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• pp.179-184
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• 2016

Bone morphogenetic protein 9 (BMP9) is a potent inducer of osteogenic differentiation of mesenchymal stem cells. The Wnt antagonist Dickkopf-1 (Dkk1) is involved in skeletal development and bone remodeling. Here, we investigated the role of Dkk1 in BMP9-induced osteogenic differentiation of MSCs. We found that overexpression of BMP9 induced Dkk1 expression in a dose-dependent manner, which was reduced by the P38 inhibitor SB203580 but not the ERK inhibitor PD98059. Moreover, Dkk1 dramatically decreased not only BMP9-induced alkaline phosphatase (ALP) activity but also the expression of osteocalcin (OCN) and osteopontin (OPN) and matrix mineralization of C3H10T1/2 cells. Furthermore, exogenous Dkk1 expression inhibited Wnt/β-catenin signaling induced by BMP9. Our findings indicate that Dkk1 negatively regulates BMP9-induced osteogenic differentiation through inhibition of the Wnt/β-catenin pathway and it could be used to optimize the therapeutic use of BMP9 and for bone tissue engineering.

• Journal of Korean Dental Science
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• 제9권1호
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• pp.9-18
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• 2016

Purpose: Wnt signaling plays an essential role in the dental epithelium and mesenchyme during tooth morphogenesis. Deletion of the Wntless (Wls) gene in odontoblasts appears to reduce canonical Wnt activity, leading to inhibition of odontoblast maturation. However, it remains unclear if autonomous Wnt ligands are necessary for differentiation of dental pulp cells into odontoblast-like cells to induce reparative dentinogenesis, one of well-known feature of pulp repair to form tertiary dentin. Materials and Methods: To analyze the autonomous role of Wls for differentiation of dental pulp cells into odontoblast-like cells, we used primary dental pulp cells from unerupted molars of Wls-floxed allele mouse after infection with adenovirus for Cre recombinase expression to knockout the floxed Wls gene or control GFP expression. The differentiation of dental pulp cells into odontoblast-like cells was analyzed by quantitative real-time polymerase chain reaction. Result: Proliferation rate was significantly decreased in dental pulp cells with Cre expression for Wls knockout. The expression levels of Osterix (Osx), runt-related transcription factor 2 (Runx2), and nuclear factor I-C (Nfic) were all significantly decreased by 0.3-fold, 0.2-fold, and 0.3-fold respectively in dental pulp cells with Wls knockout. In addition, the expression levels of Bsp, Col1a1, Opn, and Alpl were significantly decreased by 0.7-fold, 0.3-fold, 0.8-fold, and 0.6-fold respectively in dental pulp cells with Wls knockout. Conclusion: Wnt ligands produced autonomously are necessary for proper proliferation and odontoblastic differentiation of mouse dental pulp cells toward further tertiary dentinogenesis.

• 한국작물학회지
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• 제49권5호
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• pp.423-428
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• 2004

Soybean seeds contain high amounts of isoflavones that display biological effects and isoflavone content of soybean seed can vary by year, environment, and genotype. Objective of this study was to identify quantitative trait loci that underlie isoflavone content in soybean seeds. The study involved 85 $F_2$ populations derived from Korean soybean cultivar 'Kwangkyo' and wild type soybean 'IT182305' for QTL analysis associated with isoflavone content. Isoflavone content of seeds was determined by HPLC. The genetic map of 33 linkage groups with 207 markers was constructed. The linkage map spanned 2,607.5 cM across all 33 linkage groups. The average linkage distance between pair of markers among all linkage groups was 12.6 cM in Kosambi map units. Isoflavone content in $F_2$ generations varied in a fashion that suggested a continuous, polygenic inheritance. Eleven markers (4 RAPD, 3 SSR, 4 AFLP) were significantly associated with isoflavone content. Only two markers, Satt419 and CTCGAG3 had F-tests that were significant at P<0.01 in $F_2$ generation for isoflavone content. Interval mapping using the $F_2$ data revealed only two putative QTLs for isoflavone content. The peak QTL region on linkage group 3, which was near OPAG03c, explained $14\%$ variation for isoflavone content. The peak QTL region on linkage group 5, which was located near OPN14 accounted for $35.3\%$ variation for isoflavone content. Using both Map-Maker-QTL $(LOD{\geq}2.0)$ and single-factor analysis $(P{\leq}0.05)$, one marker, CTCGAG3 in linkage group 3 was associated with QTLs for isoflavone content. This information would then be used in identification of QTLs for isoflavone content with precision

• 한국식품영양과학회지
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• 제45권5호
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• pp.664-670
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• 2016

Polyglutamic acid(PGA) 함유량이 증대된 청국장이 조골세포에 미치는 영향을 세포 수준에서 관찰하고자 하였다. 조골세포에 미치는 영향을 관찰하기 위하여 세포 생존율, 염기성 인산분해효소(alkaline phosphatase, ALP) 활성, 골 석회화 형성능, 조골세포 분화 관련 유전자 발현능을 측정하였다. 세포 독성이 없는 농도에서 염기성 인산분해효소 활성을 측정한 결과 농도에 의존적으로 ALP 활성이 증가하였으며, 일반 청국장 A보다 PGA 함유량이 증가한 청국장 B가 더 높은 ALP 활성 증가 효과를 나타내었다. 청국장의 골 석회화 형성능을 측정한 결과 역시 청국장 A보다 B가 더 높은 석회화 형성능을 나타내었다. 또한 분화 관련 유전자인 OCN, OPN, Col1, ALP 역시 청국장 B가 A보다 유전자 발현이 더 높게 나타났다. 이러한 실험 결과를 바탕으로 PGA가 증대된 청국장이 기존 청국장보다 조골세포의 활성에 효과적일 것으로 생각한다.

• Tuberculosis and Respiratory Diseases
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• 제71권3호
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• pp.195-201
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• 2011

Background: Osteopontin (Opn) is recognized as an important adhesive bone matrix protein and a key cytokine involved in immune cell recruitment and tissue repair and remolding. However, serum levels of osteopontin have not been evaluated in patients with chronic obstructive pulmonary disease (COPD). Thus, the aim of this study was to evaluate and compare the serum levels of osteopontin in patients experiencing COPD exacerbations and in patients with stable COPD. Methods: Serum samples were obtained from 22 healthy control subjects, 18 stable COPD patients, and 15 COPD with exacerbation patients. Serum concentrations of osteopontin were measured by the ELISA method. Results: Serum levels of osteopontin were higher in patients with acute exacerbation than with stable COPD and in healthy control subjects ($62.4{\pm}51.9ng/mL$, $36.9{\pm}11.1ng/mL$, $30{\pm}11ng/mL$, test for trend p=0.003). In the patients with COPD exacerbation, the osteopontin levels when the patient was discharged from the hospital tended to decrease compared to those at admission ($45{\pm}52.1ng/mL$, $62.4{\pm}51.9ng/mL$, p=0.160). Osteopontin levels significantly increased according to patient factors, including never-smoker, ex-smoker and current smoker ($23{\pm}5.7ng/mL$, $35.5{\pm}17.6ng/mL$, $58.6{\pm}47.8ng/mL$, test for trend p=0.006). Also, osteopontin levels showed a significantly negative correlation with forced expiratory volume in one second ($FEV_1$%) predicted in healthy controls and stable COPD patients (r=-0.389; p=0.013). C-reactive protein (CRP) was positively correlated with osteopontin levels in patients with COPD exacerbation (r=0.775; p=0.002). Conclusion: The serum levels of osteopontin increased in patients with COPD exacerbation and tended to decrease after clinical improvement. These results suggest the possible role of osteopontin as a biomarker of acute exacerbation of COPD.