人骨髓间充质干细胞向成骨细胞分化中26RFa的作用

26RFa effects on osteogenic differentiation of human bone marrow mesenchymal stem cells

背景:研究表明26RFa在骨形成、痛觉、内分泌、心血管以及能量代谢等方面都有重要的调节作用。目的:观察26RFa在成骨培养体系中对人骨髓间充质干细胞增殖分化的影响。方法:通过MTT实验,以此确定26RFa对人骨髓间充质干细胞是否有促增殖作用及其起作用的最大活性浓度;将人骨髓间充质干细胞接种于6孔培养板上,实验分为2组:实验组含有10-11mol/L的26RFa,对照组不含26RFa。取成骨诱导8,12,16 d细胞用碱性磷酸酶试剂盒测定细胞内碱性磷酸酶活性;成骨诱导21,28 d后行茜素红染色和Von Kossa染色,计算每张玻片钙化结节数,Western blot检测cbfa1的表达。结果与结论:成骨诱导8,12,16 d后细胞内碱性磷酸酶活性实验组高于对照组(P<0.05,P<0.01,P<0.05);21,28 d后茜素红染色和Von Kossa矿化结节数实验组均高于对照组;实验组细胞cbfa1表达明显高于对照组(P<0.05)。说明在适宜的培养条件下,26RFa可促进人骨髓间充质干细胞向成骨细胞分化。

BACKGROUND：Studies have shown that 26RFa plays an important regulatory role in bone formation, pain, endocrine, cardiovascular disease and energy metabolism. OBJECTIVE：To observe the effects of 26RFa on the proliferation and differentiation of human bone marrow mesenchymal stem cells. METHODS：In order to obtain the most efficient concentration of 26RFa, human bone marrow mesenchymal stem cells were detected by 3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide analysis. cells were inoculated into 6-wel plates and then divided into two groups：experimental group treated with 10-11 mol/L 26RFa and control group with no 26RFa. After 8, 12 and 16 days of osteogenic induction, alkaline phosphatase activities in induced cells were detected using alkaline phosphatase kit. After 21 and 28 days of osteogenic induction, alizarin red staining and Von Kossa staining were performed. The number of calcified nodules over each coverslip was calculated, and the expression of cbfa1 protein was detected by western blot assay. RESULTS AND CONCLUSION：After 8, 12, and 16 days of osteogenic induction, the alkaline phosphatase activities were higher in the experimental group than the control group （P＆lt;0.05, P＆lt;0.01, P＆lt;0.05）. After 21 and 28 days of osteogenic induction, alizarin red staining and Von Kossa staining showed that the number of calcified nodules was higher in the experimental group than the control group, and the expression of cbfa1 protein was also higher in the experimental group （P＆lt;0.05）. These findings indicate that 26RFa can promote the osteogenic differentiation of human bone marrow mesenchymal stem cells under appropriate culture conditions.