杜仲含药血清诱导骨髓间充质干细胞定向分化的实验研究_药学论文.docVIP

杜仲含药血清诱导骨髓间充质干细胞定向分化的实验研究_药学论文 杜仲含药血清诱导骨髓间充质干细胞定向分化的实验研究_药学论文 作者：曾建春， 樊粤光　刘建仁，曾意荣，李显彭，易春智 【摘要】 目的研究补肾中药杜仲含药血清对骨髓间充质干细胞(BMSCs)增殖及定向分化的影响。方法将第4代MSCs以5×106接种于9块96孔板，3组，每组8孔;2块6孔板，每板3孔，2 d后换液，加入诱导液。分为空白组、地塞米松组、杜仲组。空白组加入含10%胎牛血清(FBS)的 L-DMEM培养基继续培养，地塞米松组在基础培养基加入诱导剂(含β甘油磷酸钠10mmol/L，地塞米松0.1 μmol/L，维生素C 50 mg/L)，杜仲组为含10%杜仲含药血清的L-DMEM培养基。分别自加诱导液第1天起，采用MTT法测量96孔板细胞数，每天1块板;在第10天取出1块6孔板做ALP染色;第20天做6孔板茜素红染色。结果杜仲含药血清组OD值呈现出明显的上升趋势，而空白组和地塞米松组无明显上升趋势;第10天杜仲组及地塞米松组ALP染色阳性，第13天起可见白色钙结节，第20天茜素红染色阳性。结论杜仲含药血清能促进骨髓间充质干细胞向成骨细胞分化，并刺激细胞增殖。 【关键词】 杜仲; 骨髓间充质干细胞 ; 成骨细胞 　　Abstract：ObjectiveTo study the generation and directional differentiation of BMSCs induced by Duzhong(eucommia bark ) which can invigorate the kidney. MethodsInoculated quartus generation MSCs in nine ninety six-hole cell culture plates and two six-holes cell culture plates by 5×106 cells per milliliter, all the holes were divided into three groups named blank group, DXM group and Duzhong(eucommia bark ) group , each group had eight holes or three holes, two days later, changed medium and added new medium. The medium of DXM group contained 10mmol/L β-sodium glycerophosphate, 0.1 μmol/L DXM, 50mg/L vitamin C, that of Duzhong(eucommia bark ) group contained 10% blood serum containing Duzhong(eucommia bark ). Measured cell number of 96-hole plates from the second day to the eighth day, one of the 6-holes plates was colored by AKP staining solution at the tenth day, the last one was colored by alizarin Bordeaux on the twentyth day.ResultsThe OD value of Duzhong(eucommia bark ) group rose obviously, blank group and DXM group had no obvious ascensus; on the tenth day DXM group and Duzhong(eucommia bark ) group were colored masculine; white calcium nodus could be seen on the undersurface of the holes 13days later, which could be colored by alizarin Bordeaux twenty days later. ConclusionBlood serum containing Duzhong(eucommia bark ) can induce BMSCs differentiate to osteoblast and stimulate growth of cells. 　　Key words：Duzhong(eucommi