葛根素对SD大鼠MSCs增殖及骨向分化的影响

Effect of Puerarin on Proliferation and Differentiation of Marrow Stromal Cells of SD Rats into Osteoblasts

目的:观察葛根素(Puerarin,Pue)对骨髓间充质干细胞(Marrow pluripotent stromal stem cells,MSCs)增殖和向成骨细胞(Osteoblasts,OB)分化的影响,从干细胞的角度探讨中药防治骨质疏松的可能机制和影响途径。方法:成功建立MSCs培养体系,取第三代SD大鼠MSCs,随机分为三组:空白组、不同浓度的Pue组、经典诱导组,通过MTT比色法及流式细胞仪技术,ALP染色技术观察Pue对MSCs增殖及向OB分化的影响。结果:通过对生长曲线的绘制,对多向诱导分化能力检测,及对表面抗原中CD29、CD44表达阳性,CD45表达阴性的检测,说明成功建立MSCs体外培养体系。经MTT比色法检测显示,10^(-7)mol/L、10^(-8)mol/L Pue与空白组对比具有促进MSCs增殖的作用,具有统计学意义。ALP染色阳性细胞表达率显示,10^(-6)mol/L Pue与空白组相比具有确定的诱导MSCs向OB分化的作用,说明Pue可以促进MSCs向OB分化。结论:葛根素具有促进体外培养MSCs增殖及向OB分化的能力。

Objective：To observe the effect on proliferation, differentiation to osteoblast of MSCs cultivated by Puerarin and to discuss the possible mechanism and effect of traditional Chinese medicine on prevention and treatment of osteoporosis from the view of stem cells. Methods： The cultivated system of MSCs was established successfully. The MSCs of the third generation were divided into three groups, which were blank control group, Pue group and classical induction group. The effects of Pue on proliferation and differentiation of MSCs to OB were observed by MTI？ colorimetric menthod, FCM and ALP staining technique. Results：The growth curve, the ability of multiple induction differentiation, the expression of CD29 and CtM4 and the negative ex- pression of CD45 showed that the cultivated system of MSCs in vitro was established. MTT colorimetric method showed that the comparison between 10^-7 mol/L, 10^-8 mol/L Pue and the blank control group was statistically significant, The expression of ALP staining positive cells showed that 10^-6 mol/L Pue had an effect on inducing MSCs to OB differentiation compared with the blank control group. Conclusion： Pue can promote the prolifera- tion of MSCs in vitro and the ability to differentiate into OB.