• 대한치과교정학회지
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• 제26권1호
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• pp.83-94
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• 1996

Substance P는 교정력이 가해진 치아의 치주인대 중 인장력을 받는 부위에 많이 분포하는 neuropeptide중의 하나이며, 또한 여러 조직에서 neurogenic inflammation을 야기하는 neuropeptide중의 하나로도 알려져 있다. 그러나 중요한 세포외 단백기질인 교원질의 생성에 대한 Substance P의 효과는 잘 알려져 있지 않다. 따라서 이 연구의 목적은 배양치주인대 세포에서 교원질 생성에 대한 Substance P의 효과를 평가하는 것이었다. collagenase-digestion method로 교원질 생성을 평가하였고 mRNA 수준에서 작용효과를 평가하기 위하여 Northern blot hybridization을 시행하였다. 이 연구는 또한 교원질 생성에 대한 prostaglandin과 gelatinase 생성도 포함하였으며 변성된 교원질의 분해를 평가하기 위하여 zymography를 이용하였다. 비교원성 단백질, 교원성 단백질, 상대교원질에 대한 dose-dependent effect를 보면 Substance P는 비교원성 단백질 합성을 증가시켰으나 교원성 단백질 합성은 감소시킨다. 그리하여 총 단백합성에 대한 상대적인 교원질 생성을 나타내는 상대교원질은 $7\%$에서 $3.6\%$로 감소시켰다. 세포를 indomethacin과 동시에 처리할 때 substance P의 교원질 합성 억제효과는 나타나지 않았다. 이것은 Substance P의 교원질 합성 억제효과가 prostaglandin의 생성 때문이라는 것을 의미한다. Substance P의 교원질 합성 억제효과가 procollagen mRNA의 정상(steady-state)수준에 부합하는가를 평가하기 위하여 northern blot hybridization을 시행한 결과 Substance P는 ${\alpha}1(1)$ procollagen mRNA의 양적 변동을 일으키지 않았다. Substance P의 교원질 생성 억제효과는 전사이후의 어떤 단계에서 이루어지는 현상임을 나타낸다. 치주인대세포에서 gelatinase 생성에 대한 Substance P의 효과를 알아보기 위하여 zymography를 이용하였다. zymogram 을 보면 Substance P는 치주인대세포에서 gelatinase 생성에는 아무 효과도 나타내지 않음들 알 수 있다. Substance P의 교원질 생성 억제효과가 치주인대세포에 대해 선택적인가 아닌가를알아보기 위하여 MC3T3-E1세포를 이용하였는데 Substance P는 MC3T3-E1세포의 교원질 합성에는 영향을 미치지 않았다. 이상에서 Substance P는 인간의 치주인대세포에서 교원질 합성을 억제하였다. 이 효과는 procollagen mRNA와 gelatinase 생성의 정상(steady-state) 수준의 변화 때문이 아니라 prostaglandin 생성과 연관이 있음을 알았다.

• 대한소아치과학회지
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• 제25권3호
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• pp.635-648
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• 1998

The clinical use of fluoride with a well known osteogenic action in osteoporotic patients is rational, because this condition is characterized by impaired bone formation. However, its anabolic effect has not been demonstrated well in vitro. The purpose of this study was to investigate the effects of sodium fluoride on the physiological role of osteoblastic cell. Osteoblastic cells were isolated from fetal rat calvaria. The results were as follows : 1. Mineralized nodules were shown in osteoblastic cell cultures, which had been maintained in the presence of ascorbic acid and ${\beta}-glycerophosphate$ up to 21 days. When cultures were treated with pulses of 48 hr duration before apparent mineralization was occurring, 2-fold increased in their number was detected. 2. Alkaline phosphatase activity of osteoblastic cells was inhibited by sodium fluoride in dose dependent manner. 3. The effect of sodium fluoride on the osteoblastic cell proliferation was measured by the incorporation of $[^3H]$-thymidine into DNA. As a result, sodium fluoride at $1{\sim}100{\mu}M$ increased the $[^3H]$-thymidine incorporation into DNA in a dose dependent manner. 4. The signaling mechanism activated by sodium fluoride dose-dependently enhanced the tyrosine phosphorylation of the adaptor molecule $Shc^{p66}$ and their association with Grb2, one of earlier events in a MAP kinase activation pathway cascade used by a significant subset of G protein-coupled receptors. 5. The phosphorylation of CREB(cAMP response element binding protein)was inhibited by the sodium fluoride in MC3T3E1 cells. In conclusion, the results of this study suggested that the mitogenic effect of the sodium fluoride in MC3T3E1 cell was stimulated in a dose-dependent manner and suggested "an important role for the interaction between She and Grb2" in controlling the proliferation of osteoblasts.

• Bulletin of the Korean Chemical Society
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• 제29권3호
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• pp.672-674
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• 2008

• 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.

• 대한약학회:학술대회논문집
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• 대한약학회 2003년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2-2
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• pp.124.2-124.2
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• 2003

Taurine is present in a variety of tissue and exhibits many important physiological functions in the cell. Although it is known that many tissues mediate taurine transport, its functions of taurine transport in bone have not been identified yet. In the present study, we investigated the expression of taurine transporter (TauT) and taurine uptake using mouse stromal ST2 cells and osteoblast-like MC3T3-E1 cells, which is bone related cells. Detection of TauT MRNA expression in these cells were performed by reverse transcription polymerase chain reaction (RT-PCR). (omitted)

• 농약과학회지
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• 제6권2호
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• pp.96-104
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• 2002

Paraquat의 세포독성을 알아보기 위하여 NIH 3T3 섬유모세포에 적용한 후 MTT와 NR 분석법을 이용하여 세포독성을 측정하고, paraquat의 세포독성에 대한 3-MC의 독성경감효과를 알아보기 위하여 Spraque Dawley계 수컷 랫드에 paraquat 단독 및 paraquat와 3-MC 병용투여 후 랫드의 폐를 경시적으로 채취하여 관찰하였다. Paraquat의 NIH 3T3 섬유모세포에 대한 $MTT_{50}$은 $1668.97{\mu}M$, $NR_{50}$은 $1030.25{\mu}M$로 산출되어 Borefreund와 Puemer(1984)의 독성판정기준에 의하면 저독성 물질이었다. Paraquat 단독 투여 군은 H&E 염색에서 3시간째부터 폐 모세혈관 내에 적혈구 수가 증가하기 시작하여 24시간째에는 충혈상태에 이르렀으며, 폐포사이 중격에서는 큰폐포상피수가 증가하였다. 또한 폐 조직을 둘러싸고 있는 결합조직 내에는 임파구, 대식세포 및 다형핵 백혈구 등이 다수 관찰되었고, 48시간째부터 폐포사이 중격과 폐포내에 폐포큰포식세포가 증가하기 시작하여 96시간째에는 다수의 폐포큰포식세포가 관찰되었다. Verhoeff의 iron hematoxylin 염색에서도 paraquat 단독 투여 후 24시간째에 조직변화가 가장 심하였고, 교원섬유량의 급격한 증가, 폐포의 넓이와 폐포 구멍(alveolar pore) 간격의 확장 등이 관찰되었다. 한편, paraquat와 3-MC 병용투여군은 paraquat 단독 투여 군에 비하여 조직변화가 약하게 관찰되었는데, 병용투여 후 3시간째에는 단독투여 3시간째의 소견과 유사하였으나 점차 회복되어 폐 모세혈관 내에 적혈구 수가 증가하여 24시간째에는 대조군의 구조와 거의 유사하였다. 또한 폐 조직을 둘러싸고 있는 결합조직과 임파소절에서도 paraquat 단독 투여 군에서 보였던 변화가 거의 관찰되지 않았다. Verhoeff의 iron hematoxylin 염색에서도 병용투여 후 24시간째에는 교원섬유량이 단독 투여 군에 비하여 크게 감소하였고 폐포와 폐포 구멍의 넓이도 대조군과 유사하였다.

• Journal of Ginseng Research
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• 제44권6호
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• pp.823-832
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• 2020

Background: The formation of a nanotube layer on a titanium nanotube (N-Ti) plate facilitates an active reaction between bone cells and the material surface via efficient delivery of the surface materials of the dental implant into the tissues. Studies have reported that Korean Red Ginseng extracts (KRGEs) are involved in a variety of pharmacological activities: we investigated whether implantation with a KRGE-loaded N-Ti miniimplant affects osteogenesis and osseointegration. Methods: KRGE-loaded nanotubes were constructed by fabrication on pure Ti via anodization, and MC3T3-E1 cells were cultured on the N-Ti. N-Ti implants were subsequently placed on a rat's edentulous mandibular site. New bone formation and bone mineral density were measured to analyze osteogenesis and osseointegration. Results: KRGE-loaded N-Ti significantly increased the proliferation and differentiation of MC3T3-E1 cells compared with cells on pure Ti without any KRGE loading. After 1-4 weeks, the periimplant tissue in the edentulous mandibular of the healed rat showed a remarkable increase in new bone formation and bone mineral density. In addition, high levels of the bone morphogenesis protein-2 and bone morphogenesis protein-7, besides collagen, were expressed in the periimplant tissues. Conclusion: Our findings suggest that KRGE-induced osteogenesis and osseointegration around the miniimplant may facilitate the clinical application of dental implants.

• Nutrition Research and Practice
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• 제1권1호
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• pp.29-35
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• 2007

Trace mineral studies involving metal ion chelators have been conducted in investigating the response of gene and protein expressions of certain cell lines but a few had really focused on how these metal ion chelators could affect the availability of important trace minerals such as Zn, Mn, Fe and Cu. The aim of the present study was to investigate the availability of Zn for the treatment of MC3T3-E1 osteoblast-like cells and the availability of some trace minerals in the cell culture media components after using chelexing resin in the FBS and the addition of N,N,N',N'-tetrakis-(2-pyridylmethyl)ethylenediamine (TPEN, membrane-permeable chelator) and diethylenetriaminepentaacetic acid (DTPA, membrane-impermeable chelator) in the treatment medium. Components for the preparation of cell culture medium and Zn-treated medium have been tested for Zn, Mn, Fe and Cu contents by atomic absorption spectrophotometer or inductively coupled plasma spectrophotometer. Also, the expression of bone-related genes (ALP, Runx2, PTH-R, ProCOL I, OPN and OC) was measured on the cellular Zn depletion such as chelexing or TPEN treatment. Results have shown that using the chelexing resin in FBS would significantly decrease the available Zn (p<0.05) $(39.4{\pm}1.5{\mu}M\;vs\;0.61{\pm}10.15{\mu}M)$ and Mn (p<0.05) $(0.74{\pm}0.01{\mu}M\;vs\;0.12{\pm}0.04{\mu}M)$. However, levels of Fe and Cu in FBS were not changed by chelexing FBS. The use of TPEN and DTPA as Zn-chelators did not show significant difference on the final concentration of Zn in the treatment medium (0, 3, 6, 9, $12{\mu}M$) except for in the addition of higher $15{\mu}M\;ZnCl_2$ which showed a significant increase of Zn level in DTPA-chelated treatment medium. Results have shown that both chelators gave the same pattern for the expression of the five bone-related genes between Zn and Zn+, and TPEN-treated experiments, compared to chelex-treated experiment, showed lower bone-related gene expression, which may imply that TPEN would be a stronger chelator than chelex resin. This study showed that TPEN would be a stronger chelator compared to DTPA or chelex resin and TPEN and chelex resin exerted cellular zinc depletion to be enough for cell study for Zn depletion.

• 한국치위생학회지
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• 제20권5호
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• pp.763-771
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• 2020

Objectives: Erigeron bonariensis is a type of Erigeron found throughout the tropical and subtropical areas as one of the perennial plants or pioneer plants. It is known to show detoxifying, antipyretic, and anticancer effects for various cancers. However, there are no reports on the anticancer effect of E. bonariensis on oral cancer cells. In the present study, we investigated the effects of the methanol extract of Erigeron bonariensis (MEEB) on the inhibition of cell growth and induction of apoptosis in mucoepidermoid carcinoma (MEC) cell lines, including the MC3 and YD15 oral cancer cells. Methods: MC3 Cells were treated by dimethyl sulfoxide (DMSO) or methanol extracts of 20 various natural products 20 ㎍/mL for 48 hours and cell viability were analyzed as Trypan blue exclusion assay. The effects of MEEB treatment on the cell viability of MC3 and YD15 cells, for 48 h, were analyzed by Trypan blue exclusion assay. The anticancer efficacy and apoptosis of oral cancer cell lines were analyzed by western blot analysis. The statistical significance of differences between groups was analyzed by Student's two-tailed t-test. A value of P<0.05 compared to the vehicle control was considered statistically significant. Results: Among 20 different naturally derived products, MEEB significantly inhibited cell viability and increased cleaved poly [ADP-ribose] polymerase 1 (PARP) protein in the MC3 and YD15 cells in a concentration-dependent manner. Conclusions: These results suggest that MEEB can be used as a natural anticancer drug for the treatment of human oral cancer.

• Toxicological Research
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• 제17권2호
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• pp.123-129
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• 2001

This study were carried out to investigate cytotoxicity of paraquat and bentazon that is scattering to farm products were essensial for human diet and compensatory effects of 3-methylcholanthrene (3-MC) in vitro and in vivo. In vitro, The 5.0$\times$$10^4$ cell/ml of NIH 3T3 fibroblast in each well of 24 multidish were cultured. After 24 hours, the cells were treated with solution of paraquat and bentazon (1, 25, 50, 100 pM respectively). After the NIH 3T3 fibroblast of all groups were cultured in same condition for 48 hours, Sulfohordamin B Protein (SRB) assay were performed to evaluate the cytotoxicity of cell organelles. Paraquat and bentazon $SRB_50$ were 1860.73 $\mu\textrm{M}$, 1913.38 $\mu$M respectively. In vivo, Sprague Dawley male rats divided into paraquat and bentazon only administered group and simultaneous application group of paraquat and bentazon and 3-MC. At 30 min. and 1, 3, 6, 12, 24, 48 and 96 hrs. interval after each treatment, the animals were sacrificed by decapitation and kidney were immediately removed, immersed in fixatives, and processed with routine method for light microscopic study. Paraffin sections were stained with H-E, PAM, and PAS. Under the light microscope, atrophic change of renal corpuscles were frequently observed from 3 hrs after paraquat and bentazon treatment. The increase of the mesangium was apparent from 12 hrs later after paraquat and bentazon treatment. Necrotic changes of the epithelium and loss of brush border of proximal tubules were most severe at 48 hrs after paraquat and bentazon treatment, respectively. In contrast there were no evidences of the toxic effects on renal tissues at 48hrs in paraquat and bentazon plus 3-MC treated groups.