补肾中药血清联合骨形态发生蛋白-2对人脐血间充质干细胞成骨分化的影响  

Effects of the serum of kidney-tonifying traditional Chinese herbal medicine combined with BMP-2 on osteogenic differentiation of mesenchymal stem cells derived from human umbilical cord blood

作　　者：郭杨[1] 马勇[1,2] 黄桂成[1] 潘娅岚 王建伟[3] 

机构地区：[1]南京中医药大学骨伤研究所,南京210023 [2]南京中医药大学附属医院骨伤科,南京210029 [3]南京中医药大学无锡附属医院骨伤科,无锡214071

出　　处：《中华中医药杂志》2014年第6期1809-1813,共5页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：江苏省中医药领军人才培养对象资助项目(No.LJ200924);江苏省自然基金资助项目(No.BK2011812)~~

摘　　要：目的:探讨补肾中药血清联合骨形态发生蛋白-2(BMP-2)诱导人脐血间充质干细胞定向成骨分化的可行性。方法:取第3代人脐血间充质干细胞,随机分为4组,即对照组(单纯LG-DMEM全培)、补肾中药血清组、BMP-2组及补肾中药血清组联合BMP-2组(简称联合组)。诱导后7、14、21d行碱性磷酸酶(ALP)染色,3、6、9、12d后检测ALP及骨钙素(BGP)表达,14d后Von Kossa染色测定矿化结节。结果:诱导后第14天ALP阳性表达最为明显,以联合组及BMP-2组较为显著,且联合组最优;诱导后第3、6、9、12天,联合组及BMP-2组的ALP活性与对照组相比差异有统计学意义(P<0.05,P<0.01),诱导第6天时,联合组与BMP-2组比较,差异有统计学意义(P<0.05);诱导后第9、12天,与对照组比较,联合组、BMP-2组均能提高BGP含量(P<0.05);诱导第14天后Von Kossa染色可见细胞间散在黑色颗粒,以联合组最为显著。结论:补肾中药血清联合BMP-2可有效诱导脐血间充质干细胞定向成骨分化,且具有协同效应。Objective： To discuss the feasibility of the serum of kidney-toniyfing traditional Chinese herbal medicine combined with BMP-2 inducing human umbilical cord blood mesenchymal stem cells （UCB-MSCs） into osteoblast. Methods： The third generation of UCB-MSCs was randomly divided into 4 groups： a control group, the serum of kidney-tonifying traditional Chinese herbal medicine group, BMP-2 group and the serum of kidney-tonifying traditional Chinese herbal medicine combined with BMP-2 group （combination group）. After the induction, alkaline phosphatase （ALP） staining was executed at 7d, 14d, 21d. The expression of ALP and BGP were detected at the time point of 3d, 6d, 9d, 12d. The mineralized nodules were detected by Von Kossa staining at the time point of 14d. Results： The highest positive expression rate of ALP was at the time of 14d, especially in the combination group and BMP-2 group, and higher in the combination group. There was a significant statistical difference between the combination group and control group （P〈0.05, P〈0.01） in ALP activity at 3d, 6d, 9d,12d after induction. There was a statistically significant between combination group and BMP-2 group at 6d after induction （P〈 0.05）; Compared with the control group, combination group and BMP-2 group both could increase the content of BGP at 9d, 12d after induction （P〈0.05）; Black particles were discovered scattered in cells by Von Kossa staining at the time point of 14d, the most obvious in combination group. Conclusion： The serum of kidney-tonifying traditional Chinese herbal medicine combined with BMP-2 could induce human umbilical cord blood mesenchymal stem cells.

关 键 词：补肾中药 骨形态发生蛋白-2 脐血间充质干细胞 成骨分化 

分 类 号：R285.5[医药卫生—中药学]