摘要
目的:通过观察葛根素(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)分化过程中BMP-2 mRNA表达的影响,从分子生物学角度探讨中药防治骨质疏松症的可能机制。方法:采用全骨髓贴壁法,提取MSCs,通过形态学观察及流式细胞仪等方法鉴定所提取的细胞。实验分为空白组、Pue组、经典诱导组。采用MTT法检测Pue最佳促增殖浓度;通过RT-PCR技术检测Pue对MSCs向OB分化时BMP-2 mRNA表达的影响。结果:MTT比色法检测示10-7mol/L、10^(-8)mol/L Pue能够促进MSCs增殖;ALP染色细胞阳性率显示10^(-6)mol/L Pue能够促进MSCs向OB分化;10^(-6)mol/L Pue能够促进MSCs向OB分化过程中BMP-2 mRNA的表达增强。结论:Pue具有促进体外培养MSCs增殖及提高其向OB分化中BMP-2 mRNA的表达能力。
Objective： To observe the effects of proliferation and differentiation to osteoblast of MSCs cultured by Puerarin and study the possible mechanisms and effects of traditional Chinese medicine in the prevention and treatment of osteoporosis from the view of stem cells. Methods： By using the adherence method,MSCs was extracted. By morphological observation and flow cytometry,the surface marker MSCs was identified. The experiment was divided into control group,Pue groups and classical induction group. The optimal concentration of Pue proliferation was detected by MTT method. The expression of MSCs in BMP-2 mRNA to OB differentiation was observed through RT-PCR. Results： MTT assay showed that 10-7mol / L,10-8mol / L Pue could promote the proliferation of MSCs. ALP-positive cells showed that 10-6mol / L Pue could improve the differentiation process of MSCs into OB. RT-PCR showed that 10-6mol / L Pue could improve the expression of BMP-2 mRNA in the osteogenic differentiation process of MSCs. Conclusion： Pue can promote the proliferation of MSCs in vitro and improve the expression of BMP-2 mRNA in the osteogenic differentiation process of MSCs.
出处
《上海中医药大学学报》
CAS
2016年第3期88-92,共5页
Academic Journal of Shanghai University of Traditional Chinese Medicine
基金
黑龙江中医药大学科研基金项目(201403)
