不同浓度rhBMP-2对MC3T3-E1细胞增殖、ALP活性及成骨分化的影响  

Effect of rhBMP-2 on proliferation, ALP activity and differentiation of MC3T3-E1 cells

作　　者：赵彦涛[1] 李忠海[1] 胡先同[1] 韩丽伟[1] 白玉龙[1] 李矛[1] 

机构地区：[1]解放军总医院第一附属医院骨科,北京市骨科植入医疗器械工程技术研究中心,北京100048

出　　处：《中国骨与关节损伤杂志》2016年第3期281-284,共4页Chinese Journal of Bone and Joint Injury

基　　金：国家自然科学基金(81201380);军事医学项目(13CXZ028,AWS14C007);北京市自然科学基金(7152144,7153174);北京市科技新星项目(XXJH2015102);解放军第304医院临床部重点扶持项目(2014ZD001)

摘　　要：目的研究不同浓度rh BMP-2对MC3T3-E1细胞增殖、ALP活性及成骨分化的影响规律。方法 MC3T3-E1细胞接种到培养板24 h后,试验组分别加入1.25、2.5、5、10μg/ml的rh BMP-2进行药物干预,对照组加入全培。试验组应用CCK-8法检测细胞增殖活性,PNPP法检测细胞ALP活性,RT-PCR法检测成骨标志蛋白m RNA的表达情况。结果rh BMP-2浓度为2.5μg/ml时开始促进细胞的增殖(P<0.05),rh BMP-2浓度为5μg/ml时促进作用达到峰值(P<0.05)。rh BMP-2浓度为2.5μg/ml时ALP活性显著提高(P<0.05);rh BMP-2浓度为5μg/ml和10μg/ml时,ALP活性较浓度为2.5μg/ml时继续升高,但差异无统计学意义(P>0.05)。ALP、COL1-α以及转录因子RUNX-2的m RNA含量均在加入浓度5μg/ml rh BMP-2后明显升高(P<0.05),继续加大rh BMP-2浓度,m RNA含量没有明显增加。结论在研究范围内,rh BMP-2对MC3T3-E1的影响呈浓度依赖性。Objective To study the effect of rhBMP-2 on the proliferation, Alkaline phosphatase （ALP） activity and the differentiation of MC3T3-E1 ceils. Methods The MC3T3-E1 cells were planted into 24-well cell culture plate, then the experimental groups were added with 1.25, 2.5, 5, 10μg/ml of rhBMP-2 for drug intervention, the control group was added with complete medium. CCK-8 method was used to detect the cell proliferation activity, PNPP method to detect the ALP activity, RT-PCR method to detect the mRNA expression of osteogenic protein. Results Cell proliferation activity increased until the rhBMP-2 concentration was 2.5 μg/ml （P 〈0.05）, and then reached peak value when the concentration of rhBMP-2 was 5μg/ml （P 〈0.05）; ALP activity increased when the rhBMP-2 concentration was 2.5μg/ml （P 〈0.05）, and there was no remarkable increase when the concentration of rhBMP-2 were 5 μg/ml and 10 μg/ml（P 〉0.05）. The mRNA expression of ALP, COLl-ct and RUNX-2 was significantly increased when the concentration of rhBMP-2 was 5μg/ml （P 〈0.05）, and there was no significant change when the concentration continued to increase. Conclusion In certain range, rhBMP-2 concentration- dependently affects MC3T3-E1 cells.

关 键 词：RHBMP-2 MC3T3-E1细胞 碱性磷酸酶 COL1-a RUNX-2 CCK-8 

分 类 号：R-33[医药卫生]