川续断皂苷Ⅵ对人颌骨骨髓间充质干细胞成骨分化的影响

doi:10.13591/j.cnki.kqyx.2022.03.003

摘要/Abstract

摘要： 目的探讨川续断皂苷Ⅵ(asperosaponinⅥ,ASAⅥ)对人颌骨骨髓间充质干细胞(human jaw bone marrow mesenchymal stem cells,hjBMSCs)成骨分化的影响。方法流式鉴定提取的hjBMSCs表面的特异性抗原;CCK8检测1×10^-4、1×10^-5、1×10^-6、1×10^-7、1×10^-8 mol/L的ASAⅥ药物溶液对hjBMSCs增殖的影响;用BCIP/NBT碱性磷酸酯酶显色、茜素红染色(alizarin red stain,ARS)及免疫印迹分析(western-blot,WB)验证ASAⅥ促进hjBMSCs成骨分化作用的同时,确定ASAⅥ促进hjBMSCs成骨分化合适浓度。用实时定量荧光PCR(real-time quantitative PCR,RT-PCR)进一步探索,在mRNA水平,ASAⅥ对hjBMSCs成骨分化的影响。结果实验结果表明,1×10^-4 mol/L的ASAⅥ对hjBMSCs有明显的毒性作用(P<0.01),1×10^-8～1×10^-5 mol／L对细胞增殖无明显毒性作用。与对照组相比,1×10^-5 mol／L、1×10^-6 mol／L ASAⅥ组的碱性磷酸酯酶、钙结节沉淀、成骨分化特异性蛋白表达量均明显升高。与对照组相比,1×10^-5 mol／L、1×10^-6 mol／L组的Runx2、OCN和COL-Ⅰ在mRNA水平的表达均明显上升(P<0.01),且前者略显优势。结论 1×10^-5、1×10^-6 mol／L的ASAⅥ均能较好促进hjBMSCs成骨分化,且1×10^-5 mol／L略显优势。

关键词: 川续断皂苷Ⅵ, 人颌骨骨髓间充质干细胞, 成骨分化

Abstract: Objective To investigate the effect of asperosaponinⅥ(ASAⅥ) on the osteogenic differentiation of human jaw bone marrow mesenchymal stem cells (hjBMSCs). Methods Specific antigens of hjBMSCs were identified by flow cytometry analysis. The effect of 1×10^-4, 1×10^-5, 1×10^-6, 1×10^-7, 1×10^-8 mol/L ASAⅥ on hjBMSCs' proliferation was detected by CCK8 kit; BCIP/NBT, alizarin red stain (ARS) and western-blot (WB) were used to verify the role of ASAⅥ in promoting hjBMSCs' osteogenic differentiation and to determine the appropriate concentration of ASAⅥ. The effect of ASAⅥ on the osteogenic differentiation of hjBMSCs at mRNA level was further explored by real-time quantitative PCR(RT-PCR). Results Results showed that 1×10^-4mol/L ASAⅥ had obvious toxic effects on hjBMSCs (P<0.01), while the others had no obvious side effect on cell proliferation. Alkaline phosphatase, calcium nodule precipitation, and the expression of osteogenic differentiation-specific proteins were all significantly increased in the 1×10^-5, 1×10^-6 mol/L ASAⅥ group when compared with the control group. The expression of Runx2, OCN and COL-Ⅰ at mRNA level were significantly increased in the 1×10^-5, 1×10^-6 mol/L groups compared to the control group (P<0.01), and the former was slightly predominant. Conclusion The 1×10^-5, 1×10^-6 mol/L ASAⅥ can better promote hjBMSCs' osteogenic differentiation, and 1×10^-5 mol/L group is slightly better.

Key words: asperosaponinⅥ, human jaw bone marrow mesenchymal stem cell, osteogenic differentiation

中图分类号:

R329

李燕燕, 朱珠, 谢雯静, 徐子昂, 张紫薇, 张玮. 川续断皂苷Ⅵ对人颌骨骨髓间充质干细胞成骨分化的影响[J]. 口腔医学, 2022, 42(3): 204-209.

LI Yanyan, ZHU Zhu, XIE Wenjing, XU Zi′ang, ZHANG Ziwei, ZHANG Wei. The effect of asperosaponinⅥ on human jaw bone marrow mesenchymal stem cells' osteogenic differentiation[J]. Stomatology, 2022, 42(3): 204-209.

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