• 제목/요약/키워드: MC3T3-E1

검색결과 268건 처리시간 0.023초

Protective effect of Korean Red Ginseng against glucocorticoid-induced osteoporosis in vitro and in vivo

  • Kim, Jinhee;Lee, Hyejin;Kang, Ki Sung;Chun, Kwang-Hoon;Hwang, Gwi Seo
    • Journal of Ginseng Research
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    • 제39권1호
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    • pp.46-53
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    • 2015
  • Background: Glucocorticoids (GCs) are commonly used in many chemotherapeutic protocols and play an important role in the normal regulation of bone remodeling. However, the prolonged use of GCs results in osteoporosis, which is partially due to apoptosis of osteoblasts and osteocytes. In this study, effects of Korean Red Ginseng (KRG) on GC-treated murine osteoblastic MC3T3-E1 cells and a GC-induced osteoporosis mouse model were investigated. Methods: MC3T3-E1 cells were exposed to dexamethasone (Dex) with or without KRG and cell viability was measured by the 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay. Realtime polymerase chain reaction was performed to evaluate the apoptotic gene expression; osteogenic gene expression and alkaline phosphatase (ALP) activity were also measured. Western blotting was performed to evaluate the mitogen-activated protein kinase (MAPK) proteins. A GC-induced osteoporosis animal model was used for in vivo study. Results and conclusion: The MTT assay revealed that Korean Red Ginseng (KRG) prevents loss of cell viability caused by Dex-induced apoptosis in MC3T3E1 cells. Real-time polymerase chain reaction data showed that groups treated with both Dex and KRG exhibited lower mRNA levels of caspase-3 and -9, whereas the mRNA levels of Bcl2, IAPs, and XIAP increased. Moreover, groups treated with both Dex and KRG demonstrated increased mRNA levels of ALP, RUNX2, and bone morphogenic proteins as well as increased ALP activity in MC3T3-E1 cells, compared to cells treated with Dex only. In addition, KRG increased protein kinase B (AKT) phosphorylation and decreased c-Jun N-terminal kinase (JNK) phosphorylation. Moreover, microcomputed tomography analysis of the femurs showed that GC implantation caused trabecular bone loss. However, a significant reduction of bone loss was observed in the KRG-treated group. These results suggest that the molecular mechanism of KRG in the GC-induced apoptosis may lead to the development of therapeutic strategies to prevent and/or delay osteoporosis.

방사선조사가 MC3T3-E1 조골세포주의 type I collagen과 alkaline phosphatase mRNA 발현에 미치는 영향 (The effects of irradiation on the mRNA expression of type I collagen and alkaline phosphatase in the MC3T3-E1 osteoblastic cell line)

  • 최선영;고광준
    • Imaging Science in Dentistry
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    • 제33권1호
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    • pp.51-57
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    • 2003
  • Purpose: To investigate the effects of irradiation on the phenotypic expression of the MC3T3-El osteoblastic cell line, particularly an the expression of type I collagen and alkaline phosphatase mRNA. Materials and Methods: Cells were irradiated with a single dose of 0.5, 1, 2, 4, and 8 Gy at a dose rate of 5.38 Gy/min using a cesium 137 irradiator. The specimens were then harvested and RNA extraction was carried out at 1 and 3 days after irradiation. The extracted RNA strands were reverse-transcribed and the resulting cDNA fragments were amplified by PCR. Results: The irradiated cells demonstrated a dose-dependent increase in type I collagen mRNA expression relative to the control group, with a maximum level of type I collagen mRNA expression occurring at 8 Gy. The degree of type I collagen mRNA expression increased significantly at 1 day after irradiation, but little differences were found between the control group and at the 3rd day. The amount of alkaline phosphatase mRNA expression increased significantly at land 3 days after irradiation in the 1 Gy exposed group compared with the control group. Conclusion: The amount of type I collagen and alkaline phosphatase mRNA expression increased significantly 1 day after irradiation when compared with the control group.

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까마귀쪽나무 열매 추출물의 면역증가 및 뼈 건강 효과 (Immunostimulatory and Bone Health-Promoting Activities of Litsea japonica Fruit Extract)

  • 박성진;강준철;이다영;조주현;윤민호
    • 한국식품위생안전성학회지
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    • 제35권3호
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    • pp.284-289
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    • 2020
  • 본 연구는 까마귀쪽나무 열매 추출물이 대식세포인 RAW264.7세포에서의 면역증가 효능과 조골세포인 MC3T3-E1 세포에서 뼈건강 효능에 미치는 효과를 확인하고자 하였다. 면역증가 효능을 알아보기 위해서 까마귀쪽나무열매 열수추출물(LJF-W)과 70% 에탄올추출물(LJF-70E)을 저농도(10 ㎍/mL), 중농도(100 ㎍/mL) 및 고농도(1,000 ㎍/mL)로 각각 사용하였고, 양성대조군으로는 홍삼 진세노사이드(Rg1+Rb1+Rg3 5.5 mg/g)를 이용하여 실험을 진행하였다. 실험 결과 열수추출물(LJF-W)과 70%에탄올추출물(LJF-70E) 처리군에서 NO 생성량이 무처리구 그룹에 비하여 통계적으로 유의하게 증가하였다. 염증성 cytokine인 TNF-α, IL-6, IL-1β의 생성량은 열수추출물(LJF-W)에서 무처리구에 비해 통계적으로 유의하게 증가하였으나, 70% 에탄올추출물(LJF-70E)에서는 차이가 없음을 관찰할 수 있었다. 또한 뼈건강 효능을 확인하기 위해서 까마귀쪽나무열매 추출물에 의한 조골세포인 MC3T3-E1세포의 세포증식능을 확인한 결과, 열수추출물과 70%에탄올추출물 처리군에서 조골세포 증식능이 무처리구에 비하여 통계적으로 유의하게 증가하였다. 이 결과를 통해 까마귀쪽나무열매 열수추출물(LJF-W)이 면역증가와 뼈건강에 대한 효능성분으로 가능성이 있음을 확인하였다.

Effects of Silkworm (Bombyx mori) Pupa Extract on the Function of Osteoblastic MC3T3-E1 Cells

  • Choi, Eun-Mi;Lee, Kyung-Hee;Koo, Sung-Ja
    • Food Science and Biotechnology
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    • 제14권5호
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    • pp.593-598
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    • 2005
  • Osteoporosis is recognized as one of the major hormonal deficiency diseases, especially in menopausal women and the elderly. When the estrogen level is reduced in the body, local factors, which are known to be related with bone resorption, are increased and promote osteoclastogenesis. In our previous study, we validated the estrogenicity of silkworm pupa. In this study, we investigated the effect of silkworm pupa extract (SPE) on the function of osteoblastic MC3T3-E1 cells. SPE (10 and $50\;{\mu}g/mL$) significantly elevated cell viability, alkaline phosphatase (ALP) activity, and collagen content in the cells. The effect of SPE ($50\;{\mu}g/mL$) in increasing cell viability, ALP activity, and collagen content was completely inhibited by the presence of $10^{-6}\;M$ of cycloheximide and $10^{-6}\;M$ of tamoxifen, suggesting that SPE's effect results from a newly synthesized, protein component and that it might be partly involved in estrogen action. Furthermore, we examined the effect of SPE on the $H_2O_2-induced$ apoptosis and production of local factors in osteoblasts. Treatment with SPE ($50\;{\mu}g/mL$) decreased the 0.2 mM $H_2O_2-induced$ apoptosis and the production of tumor necrosis factor (TNF)-${\alpha}$, interleukin (IL)-6 and nitric oxide (NO) in osteoblasts. Our data indicate that the enhancement of osteoblast function by silkworm pupa may prevent osteoporosis and inflammatory bone diseases.

Aqueous extract of Petasites japonicus leaves promotes osteoblast differentiation via up-regulation of Runx2 and Osterix in MC3T3-E1 cells

  • Kim, Eun Ji;Jung, Jae In;Jeon, Young Eun;Lee, Hyun Sook
    • Nutrition Research and Practice
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    • 제15권5호
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    • pp.579-590
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    • 2021
  • BACKGROUND/OBJECTIVES: Petasites japonicus Maxim (P. japonicus) has been used as an edible and medicinal plant and contains many bioactive compounds. The purpose of this study is to investigate the effect of P. japonicus on osteogenesis. MATERIALS/METHODS: The leaves and stems of P. japonicus were separated and extracted with hot water or ethanol, respectively. The total phenolic compound and total polyphenol contents of each extract were measured, and alkaline phosphatase (ALP) activity of each extract was evaluated to determine their effect on bone metabolism. To investigate the effect on osteoblast differentiation of the aqueous extract of P. japonicus leaves (AL), which produced the highest ALP activity among the tested extracts, collagen content was measured using the Sirius Red staining method, mineralization using the Alizarin Red S staining method, and osteocalcin production through enzyme-linked immunosorbent assay analysis. Also, real-time reverse transcription polymerase chain reaction was performed to investigate the mRNA expression levels of Runt-related transcriptional factor 2 (Runx2) and Osterix. RESULTS: Among the 4 P. japonicus extracts, AL had the highest values in all of the following measures: total phenolic compounds, total polyphenols, and ALP activity, which is a major biomarker of osteoblast differentiation. The AL-treated MC3T3-E1 cells showed significant increases in induced osteoblast differentiation, collagen synthesis, mineralization, and osteocalcin production. In addition, mRNA expressions of Runx2 and Osterix, transcription factors that regulate osteoblast differentiation, were significantly increased. CONCLUSIONS: These results suggest that AL can regulate osteoblasts differentiation, at least in part through Runx2 and Osterix. Therefore, it is highly likely that P. japonicus will be useful as an alternate therapeutic for the prevention and treatment of osteoporosis.