葛根素对人牙周膜干细胞成骨分化的作用

Effects of Puerarin on the Differentiation of Human Periodontal Ligament Stem Cells into Osteoblasts in Vitro

目的:探讨葛根素对人牙周膜干细胞(human periodontal ligament stem cells,hPDLSCs)成骨分化的影响。方法:向体外培养的hPDLSCs中分别加入0.01mmol/L、0.1mmol/L和1mmol/L葛根素作为实验组,另设阳性对照组和空白组。MTT法检测葛根素对hPDLSCs活性影响,采用免疫荧光法、碱性磷酸酶(alkaline phosphotase,ALP)试剂盒、茜素红染色及qRT-PCR对葛根素作用后hPDLSCs生物学特性作初步观察。结果:0.01mmol/L葛根素可明显促进hPDLSCs增殖,免疫荧光染色显示实验组I型胶原蛋白(collagen-I,COL-I)和骨桥蛋白(osteopontin,OPN)均阳性表达;与空白组对比,实验组7d后ALP活性升高,14d后茜素红染色见矿化结节形成,qRT-PCR检测ALP和骨钙蛋白(osteocalcin,OCN)分别在加药后7d和14d表达上调。结论:葛根素能够促进牙周膜干细胞向成骨细胞分化。

Objective： To assess the effects of puerarin on the differentiation of human periodontal ligament stem cells（hPDLSCs） into osteoblasts in vitro. Methods： hPDLSCs cultured in vitro in the presence of puerarin at three concentrations：0.01mmol/L,0, 1mmol/L and 1 mmol/L. The viability of hPDLSCs , immunofluorescence , alkaline phosphotase activity and mineral nodules formation were determined. In addition, the osteogenic related genes ALP and OCN were detected by RT-PCR. Results.. The viability of PDLSCs treated with puerarin at 0.01mM was significantly higher than that of control（P〈0.05）. After puerarin treamentment, immunofluorescence show that hP- DLSCs positively expressed ALP and OPN, meanwhile, alkaline phosphatase and mineral nodules were significantly increased. Quantitative real-time PCR revealed that puerarin significantly increased the mRNA expression of alka- line phosphatase （ALP）and osteocalcin（OCN）on 7th and 14th day respectively. Conclusion： Puerarin can promote the differentiation of hPDLSCs into osteoblasts.