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| 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 |
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| DOI：10.3969/j.issn.1006.7108.2016.11.004 |
| 中文关键词: 低氧 成骨生长肽 骨髓间充质干细胞 成骨分化 |
| 英文关键词:Hypoxia Osteogenic growth peptide Bone marrow mesenchymal stem cells Osteogenesis |
| 基金项目:云南省卫计委科研基金（2011WS0033) |
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| 中文摘要: |
| 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，瘦挫蓝（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lα(hypoxia inducible factor, HIF-lα)表达，培养3、7天后收集细胞，实时突光定量 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-lα蛋白，B组与D组HIF-lα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。 |
| 英文摘要: |
| Objective To investigate the effect of hypoxia on the proliferation and osteogenesis of mouse bone marrow mesenchymal stem cells ( BMSCs) induced by osteogenic growth peptide (OGP). Methods The third passage BMSCs were randomly divided into 4 groups： control group (group A, under normal oxygen condition), 1% O2 group (group B，under 1% O2 condition) , OGP group (group C，addition of OGP under normal oxygen condition) , 1% O2 combined with OGP group ( group D, addition of OGP under 1% O2 condition). 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 14d after osteogenic induction. ELISA was employed for detecting hypoxia inducible factor (HIF-lα) at 1d. qPCR was employed for detecting the mRNA of runt-related transcription factor 2 (RUNX2) and Osterix at 3d and 7d. Results 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-lα protein was not detected in group A and group C, while the expression of HIF-lα was up-regulated obviously in group B and group D (P <0. 001). Conclusion Hypoxia promotes OGP-induced BMSCs proliferation
and osteogenesis. |
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