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[旋转对大鼠培养成骨细胞骨连接蛋白和骨桥蛋白mRNA水平的影响]

[Effects of rotation on osteonectin and osteopontin mRNA level of cultured osteoblasts in rats].

作者信息

Wan Yu-Min, Ma Yong-Jie, Zhang Xiao-You, Zeng Bin, Wang Hong-Hui, Li Ying-Hui

机构信息

Institute of Space Medico-Engineering, Space Cell and Molecular Biology Laboratory, Beijing 100094, China.

出版信息

Sheng Li Xue Bao. 2005 Jun 25;57(3):384-8.

Abstract

Conditions of disuse such as bed rest, space flight, and immobilization result in decreased mechanical loading of bone, which is associated with reduced bone mineral density and increased fracture risk. Mechanisms involved in this process are not well understood except the suppression of osteoblast function. To investigate the effect of simulated weightlessness on mRNA level of extracellular matrix proteins, osteoblasts were rotated in horizontal plane as a model of simulated microgravity. Primordial osteoblasts of rats were grown for 2 d and then rotated for 24, 48 and 72 h, respectively. After isolating total RNA in cells, reverse transcription polymerase chain reaction (PT-PCR) was made to examine the mRNA level of osteopontin (OPN) and osteonectin (ON). Meanwhile, the levels of alkaline phosphatase (ALP) and osteocalcin (BGP) in the cultured medium were measured to evaluate the calcific function of cell. The expression of OPN and ON mRNA fell significantly after rotating for 24, 48 and 72 h, respectively. The contents of BGP descended significantly, meanwhile, the activity of ALP also showed a degressive tendency. Horizontal rotation decreased the expression of ON and OPN as well as diminished the secretion of BGP and ALP, which affected the calcific function of osteoblast. The results obtained suggest that depression of extracellular matrix proteins expression plays a key role in bone loss during weightlessness.

摘要

诸如卧床休息、太空飞行和固定不动等废用状态会导致骨骼的机械负荷减少,这与骨矿物质密度降低和骨折风险增加相关。除了成骨细胞功能受到抑制外,这个过程中涉及的机制尚未完全明确。为了研究模拟失重对细胞外基质蛋白mRNA水平的影响,将成骨细胞在水平面上旋转作为模拟微重力的模型。大鼠原代成骨细胞培养2天,然后分别旋转24、48和72小时。在分离细胞中的总RNA后,进行逆转录聚合酶链反应(PT-PCR)以检测骨桥蛋白(OPN)和骨连接蛋白(ON)的mRNA水平。同时,测量培养基中碱性磷酸酶(ALP)和骨钙素(BGP)的水平以评估细胞的钙化功能。分别旋转24、48和72小时后,OPN和ON mRNA的表达显著下降。BGP的含量显著下降,同时,ALP的活性也呈下降趋势。水平旋转降低了ON和OPN的表达,同时减少了BGP和ALP的分泌,这影响了成骨细胞的钙化功能。所得结果表明,细胞外基质蛋白表达的降低在失重期间的骨质流失中起关键作用。

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