摘要
[目的]观察金属Co2+、Cr3+离子对小鼠成骨细胞(MC3T3E1)的表面超微结构、凋亡、细胞周期以及在Co2+、Cr3+离子刺激下对成骨细胞分泌ALP(碱性磷酸酶)的影响。[方法]体外培养成骨细胞。扫描电镜检测细胞形态学表面超微结构,Annexin V-AITC/PI双染色法检测细胞凋亡率,流式细胞术检测细胞周期分布。ELISA(酶联免疫分析)法对培养上清液ALP含量进行检测。[结果]Co2+Cr3+金属离子明显促进MC3T3凋亡,且在24 h凋亡率最高;同时G2M分布比例明显降低,细胞停滞在G0G1期的比率明显增多;小鼠ALP Elisa试剂盒实验的OD值明显下降;扫面电镜照相显示细胞形态学表面超微结构有明显差异。[结论]Co2+、Cr3+离子对成骨细胞的细胞周期分布有明显影响,抑制其增殖使细胞大部分处于休眠期(G0G1期),同时增加其凋亡率,并可抑制成骨细胞释放ALP。
[Objective]To investigate the effect of metal ion Co2＋、Cr3＋ on surface ultrastructure 、osteoblast apoptosis,cell cycle distribution and secretion of ALP（alkaline phosphatase）. [Methods]The osteoblast was cultured in vitro.Ultrastructure of osteoblast was detected by scanning electron microscopy.The osteoblast apoptosis was detected by Annexin V-AITC/PI double staining method.Cell cycle distribution was assured by flow cytometry.ELISA method was applied to detect ALP content in serum supernatant.[Results]Co2＋、Cr3＋ metal ions significantly increased the apoptosis rate of MC3T3,especially at 24 h.At the same time,G2M（dividing phase）distribution ratio decreased and G0G1（dormancy stage） distribution ratio increased.The OD value tested by ALP ELISA method in experiment group down-regulated obviously.Scanning electron microscope photograph showed surface ultrastructure of cells had been significantly different.[Conclusion]Metal ion CO2＋,Cr3＋ have a marked effect on osteoblasts cell cycle distribution,caused most of the cell to be in dormancy stage（G0G1）.At the same time,the apoptosis rate has been upregulated and the release of ALP from osteoblast has been inhibited.
出处
《中国矫形外科杂志》
CAS
CSCD
北大核心
2011年第1期54-57,共4页
Orthopedic Journal of China
基金
国家自然科学基金资助项目(编号:30760255)
