不同分化阶段的成骨细胞对强磁重力环境的响应  

The Response to High Magneto-Gravitational Environment of Different Differentiating-Stage Osteoblasts

作　　者：胡丽芳[1] 骞爱荣[1] 李迪杰[1] 李京宝[1] 商澎[1] 

机构地区：[1]西北工业大学生命学院,西北工业大学特殊环境生物物理学研究所,空间生物实验模拟技术国防重点学科实验室,西安710072

出　　处：《中国细胞生物学学报》2013年第7期950-957,共8页Chinese Journal of Cell Biology

基　　金：国家重点基础研究发展计划(973)项目(批准号:2011CB710903);国家自然科学基金(批准号:31100667;31170812)资助的课题~~

摘　　要：研究强磁重力环境(high magneto-gravitational environment,HMGE)对不同分化阶段MC3T3-E1成骨细胞分化的影响及其可能机制。采用HMGE的μg[9 Tesla(T)]、1 g(16 T)和2 g(12 T)以及正常对照(1 g,地磁场)环境,分别对诱导成骨分化4 d和7 d的MC3T3-E1细胞处理12 h,对硝基苯磷酸法检测成骨细胞碱性磷酸酶(alkaline phosphatase,ALP)活性;茜素红S染色检测细胞矿化结节形成情况;Real-time RT-PCR检测成骨分化相关分子ALP、runt-related transcription factor 2(Runx2),I型胶原α1(type I collagenα1,Col Iα1)、骨钙素(osteocalcin,OC)以及牙本质基质蛋白1(dentin matrixprotein 1,DMP1)mRNA的表达。结果发现,经过12 h的HMGE处理后,与对照组(C)相比,μg组和1 g组成骨分化7 d的MC3T3-E1细胞ALP活性显著增加(P<0.001,P<0.01),而HMGE对成骨分化4 d细胞的ALP活性无显著影响。茜素红S染色显示,μg组和1 g组成骨分化7 d的MC3T3-E1细胞形成矿化结节较对照组增加。Real-time RT-PCR检测显示,HMGE处理12 h后,成骨分化4 d与7 d细胞成骨分化相关基因(ALP、Runx2、Col Iα1、OC和DMP1)在μg组和1 g组的表达明显上调,而2 g组下调(P<0.05,P<0.01,P<0.001)。结果表明,处于4 d与7 d不同分化阶段的MC3T3-E1细胞对12 h HMGE环境的处理敏感程度不同,7 d矿化阶段的细胞对HMGE更敏感,同时,磁场表现出促进成骨细胞分化的作用。To investigate the effects and possible mechanism of high magneto-gravitational environment （HMGE） on the differentiation of different differentiating stage MC3T3-E1 osteoblasts, the 4 d or 7 d osteogenic differentiating MC3T3-E1 osteoblasts were cultured under μg [9 Tesla（T）], 1 g （16 T）, 2 g （12 T） and control （1 g, geomagnetic field） conditions for 12 h respectively. The alkaline phosphatase （ALP） activity was analyzed by p-nitrophenyl phosphate colorimetric assay. Alizarin red s staining was used to detect mineralized nodule formation. Real time RT-PCR was applied to detect the mRNA expression of osteogenic genes including ALP, runt-relatedtranscription factor 2 （Runx2）, type I collagen al （Col Ial）, osteocalcin （OC） and dentin matrix protein 1 （DMP1）. The results showed that 12 h treatment of μg and 1 g condition significantly increased the ALP activity of 7 d differentiating MC3T3-E1 cells （P〈0.001, P〈0.01） compared with control condition, while HMGE did not affect the ALP activity of 4 d differentiating MC3T3-E1 ceils. Alizarin red S staining showed that the mineralized nodule formation of 7 d differentiating MC3T3-E1 cells increased under 11 g and 1 g condition. Real time RT-PCR results showed that the expressions of osteogenic genes （ALP, Runx2, OC, Col Ial and DMP1） were significantly up-regulated by 12 h treatment of μg and 1 g condition while down-regulated under 2 g condition （P〈0.05, P〈0.01, P〈0.001） in both 4 d and 7 d osteogenic differentiating cells. These results demonstrate that 4 d and 7 d different differentiating stage MC3T3-E1 cells show different sensitivity to 12 h treatment of HMGE while 7 d mineralizing stage cells are more sensitive to HMGE and suggest the promotion effect of magnetic field on osteoblast differentiation.

关 键 词：强磁重力环境 成骨细胞 分化 碱性磷酸酶 矿化结节 

分 类 号：R85[医药卫生—航空、航天与航海医学]