槐角提取物对颌骨骨髓间充质干细胞成骨向分化的影响

Effects of Sophora japonica extract on osteogenic differentiation of bone marrow mesenchymal stem cells

目的 :探讨槐角提取物对颌骨骨髓间充质干细胞(jaw bone marrow mesenchymal stem cells,jBMSCs)成骨分化的影响。方法:采用CCK-8法检测10、25、50、100μmol/L槐角提取物溶液对jBMSCs增殖的影响,采用BCIP/NBT碱性磷酸酯酶显色、茜素红染色检测10、25、50μmol/L槐角提取物溶液对jBMSCs成骨分化水平的影响,采用蛋白免疫印迹试验(Western blot)和实时定量荧光PCR检测槐角提取物溶液对jBMSCs成骨分化相关分子的蛋白和mRNA表达水平的影响。采用GraphPad Prism 9软件包对数据进行统计学分析。结果:100μmol/L槐角提取物溶液对jBMSCs有明显毒性作用(P<0.0001),10、25和50μmol/L浓度不影响细胞增殖(P>0.05)。与对照组相比,10、25和50μmol/L组的碱性磷酸酶活性、钙结节数量、成骨分化相关分子的蛋白和mRNA表达水平均逐渐升高。结论:10、25、50μmol/L浓度的槐角提取物能促进jBMSCs成骨向分化。

PURPOSE:To investigate the effect of Sophora japonica extract on osteogenic differentiation of jaw bone marrow mesenchymal stem cells(jBMSCs).METHODS:The effects of 10,25,50 and 100μmol/L Sophora japonica extract solutions on proliferation of jBMSCs were detected by CCK-8.BCIP/NBT alkaline phosphatase and alizarin red staining were used to detect the effect of 10,25,50μmol/L extracts of Sophora on osteogenic differentiation of jBMSCs.Western blot analysis and real-time quantitative PCR were used to detect the effects of Sophora japonica extract solution on protein and mRNA expression levels of osteogenic differentiation-related genes of jBMSCs.GraphPad Prism 9 software package was used for data analysis.RESULTS:The experimental results showed that 100μmol/L Sophora japonica extract solution had obvious toxic effect on jBMSCs(P<0.0001);while 10,25 and 50μmol/L Sophora japonica extract solution did not affect cell proliferation(P>0.05).Compared with the control group,alkaline phosphatase activity,number of calcium nodules,and protein and mRNA expression levels of osteogenic differentiation-related molecules in the 10,25 and 50μmol/L groups were gradually increased.CONCLUSIONS:The extract of Sophora japonica on 10,25,50μmol/L can enhance the osteogenic differentiation of jBMSCs.