低氧对成骨生长肽诱导的小鼠间充质干细胞成骨分化的影响  

Effect of hypoxia on the osteogenic differentiation of mouse bone marrow mesenchymal stem cells induced by osteogenic growth peptide

作　　者：张磊[1] 龚跃昆[2] 赵学凌[2] 周厚俊[3] 

机构地区：[1]云南省第一人民医院康复医学科,昆明650032 [2]昆明医科大学第一附属医院骨科,昆明650032 [3]昆明医科大学第一附属医院神经外二科,昆明650032

出　　处：《中国骨质疏松杂志》2016年第11期1380-1385,共6页Chinese Journal of Osteoporosis

基　　金：云南省卫计委科研基金(2011WS0033)

摘　　要：目的探讨低氧对成骨生长肽(osteogenic growth peptide,OGP)诱导的小鼠骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs)增殖与成骨分化的影响。方法将第三代小鼠BMSCs随机分为四组:对照组(A组,常氧条件下培养);1%O2组(B组,1%O2条件下培养);OGP组(C组,常氧条件下添加OGP培养)、1%O2联合OGP组(D组,1%O2条件下添加OGP)。分别于培养后1天、2天、3天、4天收集细胞,噻唑蓝(Methylthiazolyldiphenyl-tetrazolium bromide,MTT)比色法观察BMSCs增殖能力;培养7、14天后收集细胞,可见光比色法检测细胞碱性磷酸酶活性(alkaline phosphatase,ALP);培养1天后收集细胞,ELISA法检测低氧诱导因子1α(hypoxia inducible factor,HIF-1α)表达,培养3、7天后收集细胞,实时荧光定量PCR检测RUNX2、Osterix表达。结果 MTT结果显示,与C组相比,在培养第2、3、4天D组能明显促进BMSCs增殖(P<0.05)。与C组相比,在培养第7、14天D组能明显上调ALP表达(P<0.05)。与C组相比,在培养第3、7天D组能明显上调RUNX2、Osterix信使RNA表达(P<0.05)。在培养后1天,A组及C组未能检测到HIF-1α蛋白,B组与D组HIF-1α表达明显增加(P<0.001)。结论低氧明显增强了OGP促进BMSCs增殖及成骨分化的作用。Objective To investigate the effect of hypoxia on the proliferation and osteogenesis of mouse bone marrowmesenchymal stem cells（ BMSCs） induced by osteogenic growth peptide（ OGP）. Me thods The third passage BMSCs were randomly divided into 4 groups： control group（ group A,under normal oxygen condition）,1% O2group（ group B,under 1% O2condition）,OGP group（ group C,addition of OGP under normal oxygen condition）,1% O2 combined with OGP group（ group D,addition of OGP under 1% O2condition）. The proliferation of BMSCs was detected with methyl thiazolyl tetrazolium（ MTT） assay at 1d,2d,3d,and 4d. ALP expression was determined with visible light photocolorimetric method at 7d and 14 d after osteogenic induction. ELISA was employed for detecting hypoxia inducible factor（ HIF-1α） at 1d. q PCR was employed for detecting the mRNA of runt-related transcription factor 2（ RUNX2） and Osterix at 3d and 7d. Re sults MTT assay showed that compared with that in group C,BMSCs proliferation was promoted obviously in group D（ P〈0. 05）. Compared with that in group C,ALP expression was up-regulated obviously in group D（ P〈0. 05）. Compared with that in group C,mRNA expression of RUNX2 and Osterix increased obviously in group D. HIF-1α protein was not detected in group A and group C,while the expression of HIF-1αwas up-regulated obviously in group B and group D（ P〈 0. 001）. Conclusion Hypoxia promotes OGP-induced BMSCs proliferation and osteogenesis.

关 键 词：低氧 成骨生长肽 骨髓间充质干细胞 成骨分化 

分 类 号：R683[医药卫生—骨科学] R681[医药卫生—外科学]