模拟失重骨细胞条件培养基对成骨细胞活性的影响  

Effects of Conditioned Medium of Osteocytes Subjected to Simulated Weightlessness on Activity of Osteoblasts

作　　者：孟芮[1] 续惠云[1] 王哲[1] 张维[1] 商澎[1] 

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

出　　处：《航天医学与医学工程》2009年第4期247-251,共5页Space Medicine & Medical Engineering

基　　金：国家自然科学基金(30700152)资助

摘　　要：目的探讨三维随机回转模拟失重培养后的骨细胞条件培养基(RCM)对成骨细胞活性的调节作用。方法采用三维随机回转模拟失重培养小鼠骨细胞系MLO-Y448h后,收集回转条件培养基(RCM)和未回转对照组的条件培养基(CCM),用四甲基偶氮唑盐比色(MTT)法、对硝基苯磷酸(pNPP)法和流式细胞术(FCM)检测RCM对小鼠成骨样细胞系MC3T3-E1增殖、周期及细胞分泌碱性磷酸酶(ALP)活性的影响。结果三维随机回转48h后的MLO-Y4RCM可促进MC3T3-E1增殖,用100%的MLO-Y4RCM和CCM培养MC3T3-E148h和72h后,RCM组比CCM组分别增加了1.21和1.27倍,差异性极显著(P<0.001);周期检测结果表明,RCM可以使MC3T3-E1越过周期阻滞点(S期);MC3T3-E1的ALP活性在RCM组和CCM组之间无差异。结论三维随机回转模拟失重培养骨细胞48h后的条件培养基促进了成骨细胞的增殖;并使成骨细胞的周期越过阻滞点,对成骨细胞的ALP活性无影响。Objective To investigate the regulating effects of conditioned medium of rotation （RCM） of osteocytes subjected to simulated weightlessness on osteoblasts. Methods Conditioned medium of mouse osteocyte-like cell line MLO-Y4 was subjected to simulated weightlessness for 48 h with rondom positioning machine （RPM）.Its effects on proliferation,cell cycle and ALP activity of MC3T3-E1 were tested with MTT（methyl thiazolyl tetrazolium）,pNPP（p-Nitrophenyl Phosphate ） and flow cytometry respectively and compared with the effect of conditioned medium of control（CCM） which was not subjected to simulated weightlessness. Results After MC3T3-E1 were cultured for 48 h and 72 h,100% conditioned medium of rotation （RCM） of MLO-Y4 after simulated weightlessness for 48 h significantly stimulated proliferation of MC3T3-E1 cells as compared with conditioned medium of control （CCM）.The rate of proliferation in RCM was 1.21 and 1.27 fold as compared wtih CCM,respectively（P〈0.001）. In cell cycle assay RCM made MC3T3-E1 to overcome cell cycle arrest of S phase. There was no difference between RCM and CCM on ALP activity of MC3T3-E1 cells. Conclusion After 48 h simulated weightlessness,RCM of osteocytes promotes proliferation of osteoblasts and makes osteoblasts to overcome cell cycle arrest. There is no effects of RCM of osteocytes on ALP activity of osteoblasts.

关 键 词：失重模拟 条件培养基 骨细胞 成骨细胞 细胞增殖 细胞周期 ALP活性 

分 类 号：R851.22[医药卫生—航空、航天与航海医学]