• Journal of Ginseng Research
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• 제47권3호
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• pp.385-389
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• 2023

Background: Ginseng has been used as a traditional medicine for treatment of many diseases and for general health maintenance. Previously, we showed that ginseng did not demonstrate estrogenic property in ovariectomized mouse model. However, it is still possible that disruption of steroidogenesis leading to indirect hormonal activity. Methods: The hormonal activities were examined in compliance with OECD guidelines for detecting endocrine disrupting chemicals: test guideline (TG) No. 456 (an in vitro assay method for detecting steroidogenesis property) and TG No. 440 (an in vivo short-term screening method for chemicals with uterotrophic property). Results: Korean Red Ginseng (KRG) and ginsenosides Rb1, Rg1, and Rg3 did not interfere with estrogen and testosterone hormone synthesis as examined in H295 cells according to TG 456. KRG treatment to ovariectomized mice did not show a significant change in uterine weight. In addition, serum estrogen and testosterone levels were not change by KRG intake. Conclusion: These results clearly demonstrate that there is no steroidogenic activity associated with KRG and no disruption of the hypothalamic-pituitary-gonadal axis by KRG. Additional tests will be performed in pursuit of cellular molecular targets of ginseng to manifest mode of action.

• 한국발생생물학회지:발생과생식
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• 제8권2호
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• pp.99-111
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• 2004

난소가 제거된 생쥐를 이용하여 자궁조직에서의 ADAM-8, 9, 10, 12, 15, 17, 그리고 ADAMTS-1의 유전자의 발현이 생식호르몬에 의하여 조절되는 지를 알아보았다. 암컷 생쥐의 난소를 제거하고, 2주후에 sesame oil, 17 ${\beta}$-estradiol ($E_2$), progesterone ($P_4$ 혹은 이 둘 혼합액 ($E_2+P_4$)을 피하 주사하였다. RT-PCR 방법을 이용하여 유전자 전사체의 발현을 조사한 결과 ADAM-8, 12, 그리고 17은 oil을 주사하거나 $P_4$만을 주사한 군보다 $E_2$를 주사한 군에서 자궁조직에서의 mRNA의 양이 현저하게 증가하였다. 반면 ADAM-9, 10, 15, 그리고 ADAMTS-1은 oil을 주사하거나 $E_2$만을 주사한 군보다 $P_4$를 주사한 군에서 mRNA의 양이 현저하게 증가하였다. 또한 단백질의 발현양상의 결과도 RT-PCR의 결과와 동일하게 관찰되었다. 이러한 결과로 미루어 ADAM-8, 12, 그리고 17은 17 ${\beta}$-estradiol에 의하여, ADAM-9, 10, 15, 그리고 ADAMTS-1은 progesterone에 의하여 유전자의 발현이 upregulation 되는 것으로 생각되어진다.

• 대한한방부인과학회지
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• 제31권4호
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• pp.16-38
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• 2018

목 적: 본 연구에서는 고진음자 물 추출물의 골다공증 개선 효과를 난소적출 마우스를 이용하여 risedronate sodium(RES) 2.5 mg/kg 투여군과 비교 평가하였다. 방 법: OVX 35일 후부터 고진음자 물 추출물을 매일 1회씩 35일간 연속 경구 투여하고, 체중, 대퇴골의 중량, 골밀도, 골강도, 무기질 - 칼슘(Ca) 및 무기인(inorganic phosphorus, IP) 함량, 골량 및 구조와 골흡수에 관한 조직병리학적 변화를 혈중 osteocalcin 함량 및 bone specific alkaline phosphatase(BALP) 활성의 변화와 함께 각각 측정하였다. 본 실험에서 고진음자 물 추출물에서의 결과는 RES 경구 투여 OVX 마우스에서의 결과와 비교 평가 하였다. 결 과: OVX대조군에서는 현저한 체중 증가와 함께 대퇴골의 중량, 골밀도, 골강도, 무기질 - Ca 및 IP 함량 감소 및 지주골과 피질골의 현저한 조직병리학적 감소가 함께 확인되었으며, 혈중 osteocalcin 함량의 증가와 함께 혈중 BALP 활성의 감소가 각각 확인되었다. 이에, 전형적인 estrogen 결핍성 골다공증 소견이 OVX 수술에 의해 유발되는 것으로 관찰되었다. 한편 이렇게 OVX에 의해 유발된 estrogen 결핍성 골다공증 소견이 모든 세 용량의 고진음자 물 추출물 500, 250 및 125 mg/kg의 35일에 걸친 연속 경구 투여에 의해 투여 용량 의존적으로 현저히 억제되었고, 특히 고진음자 물 추출물 500 mg/kg은 RES 2.5 mg/kg 투여군과 비교할 만한 골다공증 개선 효과를 나타내었다. 결 론: 이상의 결과에서, 고진음자 물 추출물은 난소적출 마우스에서 투여 용량 의존적으로 유효한 골다공증 개선 효과를 나타내는 것으로 관찰되었다. 따라서 적절한 용량의 고진음자 물 추출물은 새로운 보다 효과적인 estrogen 결핍성 골다공증 개선제로서의 개발 가능성이 매우 높을 것으로 기대된다. 한편 고진음자는 15종의 약제로 구성되어 있으며, 각각의 개별 약제는 수많은 생리활성 물질을 함유하고 있어, 앞으로 생리활성을 나타내는 화학성분의 검색과 함께 다양한 방면으로 기전연구가 추가적으로 수행되어야 할 것으로 판단된다.

• Journal of Microbiology and Biotechnology
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• 제10권3호
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• pp.293-299
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• 2000

Many methods have been developed for screening chemicals with hormonal activity. Using recombinant yeasts expressing either human estrogen receptor [Saccharomyces cerevisiae ER + LYS 8127 (YER)] or androgen receptor [S. cerevisiae AR + 8320 (YAR)], we evaluated the hormonal activities of several chemicals by induction of ${\beta}-galactosidase$ activity. The chemicals were $17{\beta}-estradiol$ (E2), testosterone (T), ${\rho}-nonylphenol$ (NP), bisphenol A (BPA), genistein (GEN), 2-bromopropane (2-BP), dibutyl phthalate (DBP), di-(2-ethylhexyl) phthalate (DEHP), 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), and butylparaben (BP). To assess the estrogenicity of NP, the result of the in vitro recombinant yeast assay was compared with an E-screen assay using MCF-7 human breast cancer cells and an uterotrophid assay using ovariectomized mice. In the YER yeast cells, E2, NP, BPA, GEN, and BP exhibited estrogenicity in a doseresponse manner, while TCDD did not. All the chemicals tested, except T, did not show androgenicity in the YAR yeast cell. The sensitivity of the yeast (YER) assay system to the estrogenic effect of NP was similar to that of the E-screen assay. NP was also estrogenic in the uterotrophic assay. However, in terms of convenience and costs, the yeast assay was superior to the E-screen assay or uterotrophic assay. These results suggest that the recombinant yeast assay can be used as a rapid tool for detecting chemicals with hormonal activities.

• Journal of Genetic Medicine
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• 제19권2호
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• pp.63-75
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• 2022

Purpose: Osteoporosis is a common calcium and metabolic skeletal disease which is characterized by decreased bone mass, microarchitectural deterioration of bone tissue and impaired bone strength, thereby leading to enhanced risk of bone fragility. In this study, we aimed to identify novel genes for susceptibility to osteoporosis and/or bone density. Materials and Methods: To identify differentially expressed genes (DEGs) between control and osteoporosis-induced cells, annealing control primer-based differential display reverse-transcription polymerase chain reaction (RT-PCR) was carried out in pre-osteoblast MC3T3-E1 cells. Expression levels of the identified DEGs were evaluated by quantitative RT-PCR. Association studies for the quantitative bone density analysis and osteoporosis case-control analysis of single nucleotide polymorphism (SNPs) were performed in Korean women (3,570 subjects) from the Korean Association REsource (KARE) study cohort. Results: Comparison analysis of expression levels of the identified DEGs by quantitative RT-PCR found seven genes, Anxa6, Col5a1, Col6a2, Eno1, Myof, Nfib, and Scara5, that showed significantly different expression between the dexamethason-treated and untreated MC3T3-E1 cells and between the ovariectomized osteoporosis-induced mice and sham mice. Association studies revealed that there was a significant association between the SNPs in the five genes, ANXA6, COL5A1, ENO1, MYOF, and SCARA5, and bone density and/or osteoporosis. Conclusion: Using a whole-genome comparative expression analysis, gene expression evaluation analysis, and association analysis, we found five genes that were significantly associated with bone density and/or osteoporosis. Notably, the association P-values of the SNPs in the ANXA6 and COL5A1 genes were below the Bonferroni-corrected significance level.

• International Journal of Oral Biology
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• 제31권4호
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• pp.119-125
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• 2006

The importance of phytoestrogens to human health is currently being actively investigated. Hesperidin, abundantly found in citrus fruits, is known to possess antioxidant, anticancer, and anti-inflammatory effects. Recently, it has been reported that hesperidin inhibits bone loss and decreases serum and hepatic lipids in ovariectomized mice. In our study, to determine the possible role of hesperidin in the regulation of bone metabolism, we observed the effects of hesperidin on the proliferation and activity of osteoblasts, as well as the effects of hesperidin on osteoclast generation and activity. We observed that, when treated with hesperidin, the number and viability of osteoblastic cells increased, alkaline phosphatase (ALP) activity of osteoblastic cells increased, and osteoprotegerin (OPG) secretion from MG63 cells decreased. Hesperidin treatment had no effect on the osteoclast generation and activity in the bone marrow cell culture, but decreased the number and resorptive activity of osteoclasts generated from RAW/264.7 cells. Taken together, these results indicate that hesperidin increases the proliferation and activity of osteoblasts, while inhibiting generation and activity of osteoclasts. Although the precise role of hesperidin remains to be elucidated, our study suggests that it is one of the important modulators of bone metabolism.

• Molecules and Cells
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• 제41권6호
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• pp.575-581
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• 2018

Postmenopausal osteoporosis (PMOP) is a common systemic skeletal disease characterized by reduced bone mass and microarchitecture deterioration. Although differentially expressed SOX5 has been found in bone marrow from ovariectomized mice, its role in osteogenic differentiation in human mesenchymal stem cells (hMSCs) from bone marrow in PMOP remains unknown. In this study, we investigated the biological function of SOX5 and explore its molecular mechanism in hMSCs from patients with PMOP. Our findings showed that the mRNA and protein expression levels of SOX5 were upregulated in hMSCs isolated from bone marrow samples of PMOP patients. We also found that SOX5 overexpression decreased the alkaline phosphatase (ALP) activity and the gene expression of osteoblast markers including Collagen I, Runx2 and Osterix, which were increased by SOX5 knockdown using RNA interference. Furthermore, $TNF-{\alpha}$ notably upregulated the SOX5 mRNA expression level, and SOX5 knockdown reversed the effect of $TNF-{\alpha}$ on osteogenic differentiation of hMSCs. In addition, SOX5 overexpression increased Kruppel-like factor 4 (KLF4) gene expression, which was decreased by SOX5 silencing. KLF4 knockdown abrogated the suppressive effect of SOX5 overexpression on osteogenic differentiation of hMSCs. Taken together, our results indicated that $TNF-{\alpha}$-induced SOX5 upregulation inhibited osteogenic differentiation of hMSCs through KLF4 signal pathway, suggesting that SOX5 might be a novel therapeutic target for PMOP treatment.

• BMB Reports
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• 제52권9호
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• pp.572-576
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• 2019

Bisphosphonates are the mainstay of therapy worldwide for osteoporosis. However, bisphosphonates also have limitations. The objective of this study was to determine the role of miR-101-3p/Rap1b signal pathway in osteoclast differentiation after treatment with bisphosphonates. Our results revealed that miR-101-3p was an important regulator in bisphosphonates treated-osteoclasts. When miR-101-3p was down-regulated in bone marrow-derived macrophage-like cells (BMMs), the development of mature osteoclasts was promoted, and vice versa. However, alendronate decreased multinucleated cell number regardless of whether miR-101-3p was knocked down or over-expressed. TRAP activity assay confirmed the above results. Luciferase assay indicated that miR-101-3p was a negative regulator of Rap1b. Western blot analysis revealed that protein expression level of Rap1b in BMMs transfected with OV-miR-101-3p was lower than that in BMMs transfected with an empty vector. Rap1b overexpression increased TRAP-positive multinucleated cells, while Rap1b inhibition decreased the cell numbers. In vivo data showed that miR-101-3p inhibited osteoclast differentiation in ovariectomized mice while overexpressed of Rap1b blocked the differentiation. Taken together, our data demonstrate that miR-101-3p/Rap1b signal pathway plays a key role in osteoclast differentiation after treatment with bisphosphonates.

• 대한의용생체공학회:의공학회지
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• 제30권2호
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• pp.135-141
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• 2009

Purpose: The aim of present study is to evaluate a possibility of clinical application for the effect of low intensity ultrasound stimulation (LIUS) in mechanical characteristics of bone on osteoporotic fractures prevention. Materials and Methods: Eight virgin ICR mice (14 weeks old, approximate weight 25g) were ovariectomized (OVX) to induce osteoporosis. The right hind limbs were then stimulated with LIDS (US Group), whereas left hind limbs were not stimulated (CON Group). Both hind limbs of all mice were scanned by in-vivo micro-CT to acquire two-dimensional (2D) images at 0 week before stimulation and 3 weeks and 6 weeks after stimulation. Three-dimensional (3D) finite element (FE) models generated by scanned 2D images were used to determine quantitatively the effect of LIUS on strength related to bone structure. Additionally, distributions of Hounsfield units and elastic moduli, which are related to the bone quality, for the bones in the US and CON groups were determined to analyze quantitatively a degree of improvement of bone qualities achieved by LIUS. Results: The result of FE analysis showed that the structural strength in US Group was significantly increased over time (p<0.05), while that in CON Group was statistically constant over time (p>0.05). High values of Hounsfield units obtained from voxels on micro-CT images and high values of elastic moduli converted from the Hounsfield units were dominantly appeared in US Group compared with those in CON Group. Conclusion: These finding indicated that LIUS would improve the mechanical characteristics of osteoporotic bone via the effects of bone structure (bone strength) and quality (Hounsfield unit and elastic modulus). Therefore, the LIUS may decrease effectively the risk of osteoporotic fracture in clinics.

• Journal of Korean Neurosurgical Society
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• 제63권4호
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• pp.433-443
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• 2020

Objective : Osteoporosis is a disease of unbalanced bone metabolism that results in low bone mineral density with increased bone fragility and propensity for fractures. The increased rate of bone fracture due to osteoporosis places a significant burden on public health care expenditures. Therefore, numerous studies have been designed and performed to identify the drugs or health foods that can improve the bone quality or quantity. This study was designed to evaluate and analyze the therapeutic effects of rutin on histomorphometric values of the spine and femur in an osteoporotic mouse model induced by bilateral ovariectomy. Methods : Thirty female ICR mice (8 weeks old) underwent either a sham operation (only abdominal incision, sham group, n=10) or bilateral ovariectomy (n=20). The ovariectomized (OVX) animals were randomly divided into two groups : untreated OVX group (OVX-C, n=10), or rutin-administered group (OVX-R, n=10). The OVX-C group received weight-adjusted doses of saline vehicle and the OVX-R group received 50 mg/kg of rutin intraperitoneally, starting 1 day after surgery. At 4 and 8 weeks after surgery, serum estrogen, osteocalcin, alkaline phosphatase (ALP), and the telopeptide fragment of type I collagen C-terminus (CTX-1) were analyzed. Interleukin (IL)-1β, IL-6, IL-10, and tumor necrosis factor (TNF)-α were also analyzed. Bone histomorphometric parameters of the 4th lumbar vertebra and femur were determined by micro-computed tomography. Results : In OVX-C group, ALP, osteocalcin, CTX-1, IL-1β, IL-6, and TNF-α levels were significantly increased at 4 and 8 weeks compared to sham operation group. Rutin administration after OVX statistically significantly reduced ALP, CTX-1, IL-1β, IL-6, and TNF-α levels at 4 and 8 weeks. Rutin administration also improves bone histomorphometric parameters including trabecular bone volume fraction, trabecular thickness, and trabecular number. Trabecular separation was also decreased in OVX-R group compared to OVX-C group. Conclusion : The present study demonstrated that rutin has therapeutic effects on improving bone histomorphometric values in an OVX mouse model. The improvement in histomorphometric values may be associated with the reduction of osteoclastic activity via inhibition of IL-1β, IL-6, and TNF-α. In future studies, the mechanism for the effect of rutin on osteoporosis should be demonstrated more clearly to use rutin in human osteoporosis.