iRoot BP Plus对骨髓间充质干细胞成牙/成骨分化及自噬的影响

Effect of iRoot BP Plus on the odonto/osteogenic differentiation and autophagy of bone marrow mesenchymal stem cells

目的研究iRoot BP Plus对骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMMSCs)成牙/成骨分化的影响,初步探讨其与自噬的关系。方法全骨髓贴壁法分离培养大鼠骨髓间充质干细胞,制作iRoot BP Plus浸提液及条件培养基,Western blot筛选最佳作用浓度,碱性磷酸酶染色、qRT-PCR和Western blot观察细胞成牙/成骨相关指标的变化,Western blot检测自噬相关指标的变化。结果0.2g/L为iRoot BP Plus条件培养基的最佳作用浓度。用该浓度iRoot BP Plus条件培养基处理细胞后,细胞成牙/成骨相关指标上调。同时,自噬相关蛋白LC3Ⅱ/LC3Ⅰ比值升高,p62表达下调。结论iRoot BP Plus可促进骨髓间充质干细胞成牙/成骨分化,其机制可能与激活细胞自噬水平有关。

Objective To investigate the effect of iRoot BP Plus on the odonto/osteogenic differentiation and autophagy of bone marrow mesenchymal stem cells and preliminarily discuss its relationship with autophagy. Methods BMMSCs were harvested by the whole marrow method. iRoot BP Plus conditioned medium was made by material evaluates and the optimal concentration of iRoot BP Plus conditioned medium was determined by Western blot. ALP staining, qRT-PCR and Western blot were used to investigate the changes of odonto/osteogenic related markers of BMMSCs. The changes of autophagy related markers were detected by Western blot. Results 0.2 g/L was the optimal concentration of iRoot BP Plus conditioned medium. When treated with 0.2 g/L iRoot BP Plus conditioned medium, the expression of odonto/osteogenic markers in BMMSCs was significantly higher than that in control group, and the ratio of LC3Ⅱ/LC3Ⅰupregulated while the protein level of p62 decreased. Conclusion iRoot BP Plus can potentially promote the odonto/osteogenic capacity of BMMSCs, whose mechanism may be related with autophagy.