柚皮苷对人脐带间充质干细胞成骨分化影响.docVIP

柚皮苷对人脐带间充质干细胞成骨分化影响 　　【摘 要】目的：研究柚皮苷对人脐带间充质干细胞的生物学作用，以及对干细胞成骨分化的影响。方法：体外分离、培养及扩增HUCMSC，流式细胞技术检测HUCMSC表面特异标志物。取第3代细胞，设立2组，对照组加入地塞米松、抗坏血酸、β-甘油磷酸，柚皮苷组加入地塞米松、抗坏血酸、β-甘油磷酸、柚皮苷。观察细胞形态，进行碱性磷酸酶活性测定、钙沉积半定量分析。结果：流式细胞术检测实验中增殖培育的细胞表面白表达CD29、CD44，不表达CD34、CD45，证实了实验中培育增殖的细胞为人脐带间充质干细胞。柚皮苷组比对照组细胞液中碱性磷酸酶活性高。柚皮苷组细胞内碱性磷酸酶含量与对照组比较，差异有统计学意义（P 0.05）；茜素红染色半定量分析见柚皮苷组与对照组比较，差异有统计学意义（P 0.05）。结论：HUCMSC可以在体外采用组织块贴壁培养法培养成功，柚皮苷对HUCMSC成骨分化有促进作用。 　　【关键词】 人脐带间充质干细胞；柚皮苷；成骨分化；促进作用 　　【ABSTRACT】 Objective：To study the biological effect of naringin on human umbilical cord mesenchymal stem cell（HUCMSC）and its differentiation into osteoblasts.Methods：HUCMSC was isolated，cultured and amplified in vitro，and the surface specific markers of it were detected by flow cytometry technique.Cells of the third generation were taken to establish two groups：a naringin group and a control group，adding dexamethasone，ascorbic acid，β-glycerophosphate to the control group，adding dexamethasone，ascorbic acid，β-glycerol phosphate and naringin to the naringin group，observing the cell morphology，determining the alkaline phosphatase activity and quantitatively analyzing of calcium deposition.Results：The expression of CD29，CD44 and non-expression of CD34 and CD45 showed that the proliferation of HUCMSC was confirmed in the experiment.The activity of alkaline phosphatase was higher in the naringin group than that of the control group.The difference of the content of alkaline phosphatase between the two groups was statistically significant（P 0.05）.Alizarin red staining and semi quantitative analysis showed that there was a statistical significance between the two groups （P 0.05）.Conclusion：HUCMSC can be successfully cultured in vitro by tissue adherent culture and naringin can promote the differentiation of HUCMSC into osteoblasts. 　　【Keywords】 human umbilical cord mesenchymal stem cells；naringin；osteogenic differentiation； 　　promoting effect 　　关节置换手术中，处理骨缺损一直是个难题，阻挡了关节外科的发展。对于髋膝关节置换中出现的骨缺损的处理，由同种异体骨、自体骨移植发展到3D打印骨缺损来修补[1]。