小寡核苷酸对大鼠骨髓间充质干细胞向成骨细胞分化的影响

Effect of oligodeoxynucleotide on differentiation of rat bone marrow mesenchymal stem cells to osteoblasts

目的:筛选具有促进大鼠骨髓间充质干细胞(BMSCs)成骨分化作用的小寡核苷酸(ODN),探讨ODN对BMSCs向成骨细胞分化的影响。方法:取第3代BMSCs孵育24h后进行成骨诱导,双盲法加入12条不同的ODN,PBS作为溶媒对照组,应用碱性磷酸酶(ALP)试剂盒检测ODN于不同工作浓度(4.0、2.0、1.0和0.5mg.L-1)、不同时间点(24、72和120h)刺激经成骨诱导后的BMSCs内ALP的表达水平,筛选出具有促成骨分化作用的ODN。结果:与PBS溶媒对照组比较,ODN工作浓度为4.0和0.5mg.L-1时,实验组与对照组ALP表达水平差异无显著性(P>0.05),ODN工作浓度为2.0和1.0mg.L-1时,有2条ODN在不同时间点(24、72和120h)均可促进BMSCs内ALP的表达(P<0.01)。结论:在12条不同的ODN中共筛选出2条具有促进BMSCs成骨分化作用的ODN,表明特定序列的ODN能够影响大鼠BMSCs向成骨细胞的分化。

Objective To investigate the effect of oligodeoxynucleotide（ODN） on differentiation of rat bone marrow mesenchymal stem cells（BMSCs） to osteoblasts.Methods BMSCs at the third generation were incubated for 24 h before 12 different ODN were added.The levels of alkaline phosphatase（ALP）,reflecting the degree of osteogenic differentiation were detected in four replicate wells of BMSCs stimulated with each ODN with different concentrations（4.0,2.0,1.0 and 0.5 mg·^L-1） at different time points（24,72 and 120 h）.Results There were no significant differernces of the mean level of ALP between 4.0 and 0.5 mg·L^-1 ODN groups and PBS control group;but in 2.0 and 1.0 mg·L^-1 ODN groups,there were two ODN which could increase the ALP levels at different time points（24,72 and 120 h）（P〈0.01）.Conclusion Two ODN of 12 different ODN can promote the diffentiation of BMSCs to osteoblasts.It suggests that ODN with special sequence characteristic can significantly promote the differentiation of rat BMSCs to osteoblasts.