前列腺素E2调控骨髓间充质干细胞成骨分化最佳浓度及其效应基因

背景：前列腺素E2不仅是参与机体炎症反应的炎性递质，而且在骨组织形成与改建过程中起着非常重要的调节作用，且前列腺素E2在其他干细胞如神经干细胞、胚胎干细胞的增殖分化中，也表现出类似的调节作用。目的：研究前列腺素E2对骨髓间充质干细胞增殖与骨向分化的影响，检测其促增殖和成骨分化的最佳剂量，筛选出前列腺素E2的效应基因，评价前列腺素E2诱导成骨的作用机制。方法：提取SD大鼠骨髓间充质干细胞并鉴定，配制10-2，10-3，10-4，10-5，10-6g／L不同质量浓度的前列腺素E2溶液，分别与第3代骨髓间充质干细胞共培养，空白组为含体积分数10％胎牛血清的L．DMEM。M丌比色法及碱性磷酸酶定量检测，筛选出前列腺素E2促骨髓间充质干细胞增殖及成骨分化的最佳剂量。利用基因芯片筛选出最佳成骨诱导浓度前列腺素E2诱导后14d的骨髓间充质干细胞与未诱导组的差异表达基因。结果与结论：不同质量浓度前列腺素E2诱导实验数据进行统计学分析后发现前列腺素E2最佳促骨髓间充质干细胞增殖浓度为1×10-4g／L，前列腺素E2最佳促骨髓间充质干细胞成骨分化浓度为1×10-5g／L。基因芯片筛选发现Cxcll3、Cd55基因及PPAR、MAPK信号通路可能与前列腺素E2促骨髓间充质干细胞向成骨分化关系密切。

Optimal concentration of prostaglandin E2 for osteogenic differentiation of bone marrow mesenchymal stem cells and its effectors

BACKGROUND： Prostaglandin E2 is not only involved in the body＇s inflammatory response as an inflammatory mediator, but also plays a very important role in the process of bone formation and remodeling. In addition, prostaglandin E2 also regulates the proliferation and differentiation of other stem cells, such as neural stem cells and embryonic stem cells. OBJECTIVE： To observe the effects of prostaglandin E2 on proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells and to confirm the best concentration of prostaglandin E2, then to screen differentially expressed genes by gene chip technology, and to study the mechanism of prostaglandin E2 in the osteoinductive process of bone marrow mesenchymal stem cells.METHODS： Bone marrow mesenchymal stem cells were isolated and cultured from Sprague-Dawley rats, and prostaglandin E2 was prepared to different concentrations of solutions： 103, 10-3, 10~, 10~ and 10-6 g/L. Then, passage 3 cells were cultured in prostaglandin E2 solutions, and low-glucose Dulbecco＇s modified Eagle＇s medium containing 10% fetal bovine serum. MTT colorimetric assay and quantitative detection of alkaline phosphatase were used to confirm the best concentration of prostaglandin E2 promoting proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells. After 14 days, the gene-chip technique was used to detect the differentially expressed genes. RESULTS AND CONCLUSION： The experimental data were statistically analyzed, we found that the best concentration of prostaglandin E2 for the proliferation of bone marrow mesenchymal stem ceils was I x 104 g/L, and the best concentration of prostaglandin E2 for the osteogenic differentiation of bone marrow mesenchymal stern cells was 1 xl0-5 g/L. According to the gene chip, Cxc113, Cd55 gene and PPAR, MAPK signaling pathways might be the key genes and pathways in the osteogenic differentiation process of bone marrow mesenchymal stem cells.