摘要
目的探讨极度低氧条件下Ckip-1 siRNA促进骨髓间充质干细胞(BMSCs)成骨分化作用。方法差速贴壁法分离培养兔BMSCs,常氧(20%)及极度低氧(1%)条件下培养72 h,实时荧光定量PCR(qPCR)及Western blot检测成骨基因Run相关转录因子2(Runx2)、SMAD5及骨形态发生蛋白2(BMP2)表达,培养3、5、7 d后检测上清中碱性磷酸酶(ALP)活性。再将BMSCs置于1%氧浓度下培养72 h,观察Ckip-1 siRNA对Runx2、SMAD5及BMP2表达及ALP活性的影响。结果 1%氧浓度下BMSCs的成骨基因表达及ALP活性明显低于20%氧浓度下,差异有统计学意义(P <0.05);Ckip-1 siRNA转染后BMSCs成骨基因表达及ALP活性明显高于未转染,差异有统计学意义(P <0.05)。结论极度低氧可抑制BMSCs成骨分化能力,而Ckip-1 siRNA促进BMSCs在极度低氧条件下向成骨细胞分化。
Objective To investigate the osteogenic differentiation of bone marrow mesenchymal stem cells(BMSCs)by Ckip-1 siRNA under extreme hypoxia.Methods BMSCs were isolated and cultured in normal oxygen(20%)and extreme hypoxia(1%)for 72 h.The expression of osteogenic gene Runt-related transcription factor 2(Runx2),Smad5 and bone morphogenetic protein 2(BMP2)were detected by real-time quantitative PCR(qPCR)and Western blot.Alkaline phosphatase(ALP)activity in the supernatant was detected after 3,5,7 days of culture.BMSCs were cultured in 1%oxygen for 72 h to observe the effect of Ckip-1 siRNA on the expression of Runx2,Smad5,BMP2 and the activity of ALP.Results The osteogenic gene expression and ALP activity of BMSCs in 1%oxygen concentration were significantly lower than those in 20%oxygen concentration group,and the differences were statistically significant(P<0.05);the osteogenic gene expression and ALP activity of BMSCs after transfection with Ckip-1 siRNA were significantly higher than those without transfection,and the differences were statistically significant(P<0.05).Conclusion Hypoxia can inhibit the osteogenic differentiation of BMSCs,and Ckip-1 siRNA can promote the differentiation of BMSCs into osteoblasts.
作者
刘想忠
宁宇
胡静
杨傲飞
蔡寒涛
李章华
LIU Xiangzhong;NING Yu;HU Jing;YANG Aofei;CAI Hantao;LI Zhanghua(Department of Orthopaedics,Zhongnan Hospital of Wuhan University,Hubei Province,Wuhan 430000,China;Department of Orthopaedics,Tongren Hospital of Wuhan Universyity,Hubei Province,Wuhan 430000,China;Department of Orthopaedics,Xiangyang Hospital of Traditional Chinese Medicine,Hubei Province,Xiangyang 441000,China;Graduate School,Wuhan Sports University,Hubei Province,Wuhan 430000,China;Department of Orthopaedics,Hubei Provincial Hospital of Traditional Chinese Medicine,Hubei Province,Wuhan 430000,China;the First Clinical School,Hubei University of Traditional Chinese Medicine,Hubei Province,Wuhan 430000,China)
出处
《中国医药导报》
CAS
2020年第18期10-14,23,共6页
China Medical Herald
基金
国家自然科学基金资助项目(81472103)
湖北省武汉市卫生和计划生育委员会科研项目(WX18M01)
湖北省武汉市科技局应用基础前沿项目(2019020701011471)。
