氯化锂对人颌骨来源的骨髓间充质干细胞增殖及骨向分化能力的影响

Effects of lithium chloride on the proliferation and osteogenic differentiation of human orofacial bone marrow mesenchymal stem cells

目的:探讨氯化锂(LiCl)对颌骨来源的骨髓间充质干细胞(OF-BMMSCs)增殖及骨性分化的影响。方法:采用有限稀释法克隆化培养人颌骨来源的骨髓间充质干细胞;正常培养基中加入不同浓度LiCl(5、10、20、40 mmol/L),同时设立对照组(0 mmol/L);运用MTT法分析不同浓度LiCl对人OF-BMMSCs增殖活性的影响;采用组织化学染色法、RT-PCR法测定不同浓度LiCl处理人OF-BMMSCs后的ALP活性及相关骨向分化基因(ALP,Runx-2,OCN)的表达。结果:5 mmol/L LiCl促进OF-BMMSCs增殖,对ALP、Runx-2、OCN表达的影响作用不明显(P>0.05),20和40 mmol/L LiCl抑制细胞增殖(P<0.05),促进ALP、Runx-2、OCN基因表达(P<0.05)。结论:LiCl能够促进人OF-BMMSCs的骨向分化。

Objective： To investigate the effects of lithium chloride （LiC1） on the proliferation and osteogenic differentiation of human orofacial-derived bone marrow mesenchymal cells （OF-BMMSCs）in vitro. Methods： Human OF-BMMSCs were isolated and cultured by limited dilution method. The cloned OF-BMMSCs were stimulated with LiC1 at different concentrations（0, 5, 10, 20, 40 retool/L） respectively. After 3 days stimulation, proliferation of OF-BMMSCs was examined by MTF assay. The osteogenic differentiation of OF- BMMSCs was evaluated by Alizarin red staining, ALP staining and real-time quantitative PCR for the mRNA expression of ALP, Runx- 2 and OCN after treatment by LiC1 and followed by osteogenic induction. Results：LiC1 at 5 mmol/L promoted the proliferation of OF- BMMSCs（P 〈 0.05）, but showed no significant effect on the mRNA expression of ALP, Runx-2 and OCN of OF-BMMSCs（P 〉 0.05 ）. However, LiC1 at 20 and 40 mmol/L inhibited the cells proliferation （P 〉 O. 05 ）, while, promoted the mRNA expression of the genes（P 〈 0.05 ）. Conclusion： LiC1 may promote the osteogenic differentiation of OF-BMMSCs.