니코틴이MG63조골세포주의오스테오칼신과오스테오프로테제린의 생성및mRNA 발현에 미치는영향
Effects of nicotine on the formation of osteocalcin and osteoprotegerin and synthesis of its mRNA in MG63 osteoblast-like cell
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흡연이 골형성 세포의 기능을 억제한다는 것은 이미 잘 알려져 있으나 대다수의 연구가 주로 역학조사에 기초하여 이루어져 왔다. 본 연구의 목적은 흠연과 골형성 세포의 상관관계를 실험적으로 평가하기 위한 것이다. 본 연구를 위해 인간 골육종 세포인 MG63 조골세포주를 이용하였으며, 니코틴이 조골세포의 증식, 염기성 인산분해효소의 활성, 오스테오칼신과 오스테오프로테제린의 생성 그리고 mRNA 발현에 미치는 영향을 관찰하였다. MG63 조골세포를 니코틴 농도가 각각 1.0 μＭ, 1.0 mM, 2.5 mM, 5.0 mM, 7.5 mM, 그리고 10.0 mM,이 함유된 배지에서 1일, 2일, 3일 그리고 6일동안 배양 실험을 하여 다음과 같은 결과를 얻었다. MG63 조골세포의 증식이 저농도의 니코틴에서는 일시적으로 활성화되었지만 5.0 mM 이상의 고농도에서는 억제되었다. 염기성 인산분해효소의 활성은 니코틴 농도가 증가함에 따라 감소하였다. 오스테오칼신 생성양은, 배양 1일 후, 5.0mM 이하의 농도에서 증가하였으나 7.5 mM 이상의 고농도에서는 감소하였다. MG63 조골세포를 3일 배양한 경우, 오스테오칼신 생성양은 1.0 mM의 저농도에서도 감소하였다. 니코틴이 오스테오칼신 단백질 생성과 오스테오칼신 mRNA 생성에 미치는 영향은 1일과 3일에서 다소 차이가 있었다. 오스테오프로테제린의 생성양은 모든 실험군에서 니코틴을 함유하지 않은 대조군보다 감소하였다. 하지만 mRNA 수치는 단백질 생성과 상반되게 7.5 mM (3일), 5.0 mM (6일) 이상의 고농도에서 증가하였다.
Abstract
The purpose of this study was to evaluate the correlation between nicotine and the activity of bone forming cell. MG63 osteoblast-like cells were used for this study. Several factors were examined including the proliferation of cell, alkaline phosphatase activity, the formation of osteocalcin and osteoprotegerin, and the synthesis of its mRNA. MG63 osteoblast-like cells were incubated for 1, 2, 3 and 6 days with nicotine added to the culture medium in 1.0 μM, 1.0 mM, 2.5 mM, 5.0 mM, 7.5 mM, and 10.0 mM concentrations. The proliferation of MG63 osteoblast-like cells was temporarily activated at the low nicotine concentrations. At high concentrations (>5.0 mM), however, it was suppressed. Alkaline phosphatase activity was suppressed in a dose-dependent manner as the concentration of nicotine increased. Osteocalcin decreased in a dose-dependent manner at high nicotine concentrations of more than 7.5 mM and the same result was show when the osteoblasts were treated with low concentrations for longer than 3 days. There was a difference in the influence of nicotine on the synthesis of osteocalcin mRNA and formation of osteocalcin itself at 1 and 3 days. Generally, osteoprotegrin notably declined in all experimental groups. However, the level of its mRNA inc-reased at high nicotine concentrations of more than 7.5 mM after 3 days and more than 5.0 mM after 6 days.
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