促小鼠红细胞生成素对体外培养小鼠成骨细胞成骨作用的实验研究  

The Effect of rm EPO on Cell Osteogenic Activity on Mice Cultured in vitro

作　　者：廖菲[1] 徐涛涛[1] 肖鲁伟[1] 童培建[1] 吴承亮[1] 

机构地区：[1]浙江中医药大学,杭州310053

出　　处：《浙江中医药大学学报》2015年第6期473-477,504,共6页Journal of Zhejiang Chinese Medical University

基　　金：国家自然科学基金(81273770)~~

摘　　要：[目的]探究促小鼠红细胞生成素(recombinant mouse Erythropoietin,rm EPO)对体外培养小鼠成骨细胞成骨活性的影响。[方法]3d内新生C57乳鼠10只,无菌条件下酶消化法分离其成骨细胞。原代培养成骨细胞传第一代,将生长良好的成骨细胞制成细胞悬液,随机分为空白组和rm EPO组(10ng·m L-1)。MTT比色法检测细胞增殖;比色法检测细胞碱性磷酸酶(alkaline phosphatase,ALP)活性;用茜素红染色观察细胞外基质矿化情况;用RT-PCR法检测细胞成骨相关基因(ALP、OPG、RANKL)的表达水平。[结果]rm EPO能够显著促进细胞增殖(与空白组相比P<0.05)、细胞ALP活性的表达(与空白组相比P<0.05)、细胞外基质矿化。能够显著上调ALP、OPG基因表达(与空白组相比P<0.05),下调RANKL基因的表达(与空白组相比P<0.05)。[结论]rm EPO能促进小鼠成骨细胞成骨活性。[Objective] To investigate the role of rmEPO on cellosteogenic activity of osteoblast cells. [Methods] 10 C57 mice（3d old） were used, and the osteoblast cells were separated by enzyme digestion under the aseptic condition. Primary osteoblast cells were cultured to obtain the first generation of the cells, then divided into normal group and rmEPO group（10ng·min＆-1 rmEPO treated）. We used the MTT colorimetric method to test cell proliferation assay, the colorimetric method to test the ALP activity, the alizarin red staining method to observe the calcified area, RT-PCR to test the expression of osteogenic genes（ALP,OPG,RANKL genes）.[Resuhs] After the rmEPO treatment, osteoblast cells showed increased cell viability compared with the normal group（P〈0.05）. And rmEPO-treated cells showed enhanced ALP activity and enhanced mineralization（P〈0.05）, rmEPO treatment also significantly promoted ALP and OPG expression（P〈0.05） while significantly inhibiting RANKL expression（P〈0.05）. [Conclusion] rmEPO can induce the cellosteogenic activity of mouse osteoblast cells.

关 键 词：促红细胞生成素 成骨细胞 成骨活性 碱性磷酸酶 基因表达 

分 类 号：R331[医药卫生—人体生理学]