硫酸钙对人骨髓基质干细胞成骨过程中VDR表达的影响

目的 观察硫酸钙对入骨髓基质干细胞(hBMSC)成骨诱导过程中维生素D受体(VDR)基因表达的影响.方法 原代分离培养hBMSC,在倒置显微镜下观察细胞形态,流式细胞仪检测hBMSC表型,取第2代hBMSC,分为硫酸钙组与对照组,分别用含或不含硫酸钙的成骨诱导液进行成骨诱导,诱导培养14 d后收集细胞,实时定量PCR检测两组VDR mRNA的表达水平.结果 流式细胞仪检测hBMSC表面抗原,CD34、CD45表达阴性,CD29、CD71、CD106表达阳性,阳性率分别为98.74％、65.74％、68.46％.与对照组相比,硫酸钙组倒置显微镜观察细胞呈长梭形,细胞两极成束状排列,细胞增殖变快,细胞呈多个散在的细胞结节,并且产生大量基质成分.实时定量PCR结果显示,与对照组相比,硫酸钙组VDR mRNA的相对表达明显增加(2.64±0.49)比(0.99 ±0.16),t=6.297,P<0.01].结论 在hBMSC成骨诱导过程中,硫酸钙能够促进VDR基因表达.

Effects of calcium sulfate on the expression of VDR mRNA during the osteogenesis of human bone marrow stromal stem cell

Objective To investigate the effect of calcium sulfate on the expression of vitamin D receptor(VDR) gene in human bone marrow stromal stem cells (hBMSC) during the osteogenic induction.Methods Primary isolated hBMSC were cultured and observed under an inverted microscope.Flow cytometry was used to identify the phenotype of hBMSC, and the second generation of hBMSC were divided into calcium sulfate group and control group.Cells were cultured in osteogenic induction medium with or without calcium sulfate and were harvested after 14 days.Real time quantitative polymerase chain reaction was used to detect the expression of VDR mRNA in both groups.Results The surface antigen of hBMSC was detected by flow cytometry.The expression of CD34 and CD45 were negative;the expression of CD29, CD71 and CD106 were positive, and the positive rate was 98.74％, 65.74％, or 68.46％, respectively.Compared with control group, under inverted microscope, cells in calcium sulfate group were spindle-shaped, and arranged in bundles;the proliferation were accelerated, and more scattered cell nodules and a large amount of matrix components were found in calcium sulfate group.Quantitative real-time PCR results showed that compared with control group, the relative expression of VDR mRNA in calcium sulfate group was increased(2.64±0.49) vs.(0.99±0.16),t=6.297, P<0.01].Conclusion Calcium sulfate can promote the expression of VDR mRNA in hBMSC during osteogenic induction.