ProgranuIin衍生工程蛋白Atsttrin对牙周膜干细胞成骨分化作用的影响

Effects of Progranulin Derived Engineered Protein Atsttrin on Osteogenic Differentiation of Periodontal Ligament Stem Cells

目的:探讨Atsttrin对牙周膜干细胞(periodontal ligament stem cells,PDLSCs)成骨分化的促进作用及Atsttrin逆转肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)对PDLSCs成骨分化的抑制作用。方法:体外分离培养PDLSCs,实验组分别加入25 μg/L Atsttrin、10 μg/L TNF-α、10 μg/L TNF-α+25 μg/L Atsttrin,检测PDLSCs的碱性磷酸酶(alkaline phosphatase activity,ALP)活性及成骨相关标志物ALP、runt相关转录因子-2(runt-related transcription factor-2,Runx2)的mRNA及蛋白表达水平;21 d时,进行茜素红染色和钙含量测定。结果: Atsttrin增强PDLSCs的ALP活性,增强ALP、Runx2基因及蛋白表达水平,促进矿化结节的形成。在各时间点中, 25 μg/L Atsttrin组促进PDLSCs成骨分化及矿化的能力最强。结论: Atsttrin能促进PDLSCs的成骨分化及矿化,能拮抗TNF-α对PDLSCs成骨分化的抑制作用。

Objective: To explore the positive effect of Atsttrin on osteogenic differentiation of PDLSCs and the reversing effect of Atsttrin on TNF-α-inhibited PDLSCs osteogenic differentiation.Methods: PDLSCs were obtained and treated with 25 μg/L Atsttrin,10 μg/L TNF-α,and 10 μg/L TNF-α+ 25 μg/L Atsttrin,respectively.Alkaline phosphatase activity (ALP) and gene/protein expression levels of ALP and runt-related transcription factor-2 (Runx2) were assessed.The formation of mineralized nodules and determination of calcium content was observed.Results: Atsttrin enhanced the ALP activity and osteogenesis gene/protein expressions of PDLSCs,and promoted the formation of mineralized nodules.At three time points,the highest osteogenic differentiation and mineralization ability of PDLSCs were found in the 25 μg/L Atsttrin group.Conclusion: Atsttrin can directly promote the osteogenic differentiation and mineralization of PDLSCs,and can reverse the inhibitory effect of TNF-α on osteogenic differentiation of PDLSCs.