摘要
目的探讨乳鼠成骨细胞碱性磷酸酶活性的检测方法,为研究骨代谢异常及骨质疏松症等骨病提供简便可靠的检测手段。方法采用经体外培养的乳鼠成骨细胞,制备上清液、裂解液和冻融液样本,分别用二乙醇胺、2-乙基氨基乙醇、2-氨基-2-甲基-1-丙醇3种不同缓冲液的试剂方法,测定其碱性磷酸酶活性,并作比较分析。结果使用二乙醇胺缓冲液的试剂方法,对3种标本的测定值都最高,与各组比较分析,差异均有显著性(P<0.01),且变异系数最小,其他两种方法的测定结果无明显差异(P>0.05)。用3种不同缓冲液试剂方法检测3种不同方法制备的标本,均以裂解液中碱性磷酸酶活性最高,冻融液次之,上清液最低,分析结果,差异均有显著性(P<0.01)。结论用二乙醇胺缓冲液的试剂方法,测定体外培养乳鼠成骨细胞裂解液碱性磷酸酶的活性,效果较为理想。
[Objective] To study the methodology on the determination of osteoblast alkaline phosphatase in sucking rat in order to provide a simple and reliable means for bone diseases, such as bone metabolic abnormality and osteoporosis etc. [Method] The osteoblast from sucking rat was cultured in vitro to obtain the samples of supernatant, spliting liquid, freezing-melting liquid. The alkaline phosphatase in samples above were determined using the different buffer solutions of diethanol, 2-ethylaminothanol and 2-amino-2-methyl-l-propanol and the results were analyzed. [Results] The activities of alkaline phosphatase in three types of samples above and precision were the highest in diethanol buffer solution and had obvious differences as compared to other two groups （P 〈0.01）, but the results of other two methods had no difference （P 〉0.05）. In three types of samples, the results of alkaline phosphatase were different and had statistical diversity by the three methods（P 〈0.01）, the highest in spliting liquid and the lowest in supernatant. [Conclusion] The method using diethanol as buffer solution is more ideal for the determination of osteoblast alkaline phosphatase activity in cultures of sucking rat in vitro.
出处
《中国现代医学杂志》
CAS
CSCD
北大核心
2006年第10期1495-1497,1500,共4页
China Journal of Modern Medicine
关键词
乳鼠
成骨细胞
碱性磷酸酶
测定
sucking rat
osteoblast
alkaline phosphatase
determination
