熊果酸对小鼠成牙骨质细胞系OCCM-30细胞分化的促进作用

doi:10.13481/j.1671-587x.20190502

摘要/Abstract

摘要： 目的：探讨熊果酸对小鼠成牙骨质细胞系OCCM-30细胞分化的影响，为牙根吸收的修复提供理论依据。方法：将对数生长期的OCCM-30细胞分为对照组和不同浓度熊果酸组。其中熊果酸组OCCM-30细胞以3种浓度（0.625、1.250和2.500 μmol·L^-1）的熊果酸处理，对照组不做任何处理。采用MTT法检测不同时间点（24、48和72 h）各组OCCM-30细胞增殖抑制率，碱性磷酸酶（ALP）法检测细胞体外分化和矿化情况，实时定量PCR法检测24h内骨桥蛋白（OPN） mRNA表达水平，Western blotting法检测给药3和5 d时OPN蛋白表达水平。结果：不同浓度熊果酸组与对照组OCCM-30细胞增殖抑制率比较差异无统计学意义（P>0.05）。与对照组比较，2.500μmol·L^-1熊果酸组熊果酸处理3、5和7 d后，OCCM-30细胞中ALP活性明显升高（P<0.05），不同浓度熊果酸组OCCM-30细胞中OPN mRNA和蛋白表达水平明显升高（P<0.01）。结论：一定浓度熊果酸对小鼠成牙骨质细胞系OCCM-30细胞增殖无明显抑制作用，但可促进其分化并上调OPN mRNA和蛋白表达水平。

关键词: 熊果酸, 成牙骨质细胞, 细胞增殖, 细胞分化

Abstract: Objective:To study the effects of ursolic acid on the differentiation of cementoblast cell line OCCM-30 cells, and to provide the theoretical basis for the repair of root restoration. Methods:The OCCM-30 cells in logarithmic growth phase were divided into control group and different concentrations of ursolic acid groups. The OCCM-30 cells in ursolic acid groups were treated with different concentrations(0.625, 1.250 and 2.500 μmol·L^-1) of ursolic acid and the cells in control group did not receive any treatment. MTT method was performed to detect the inhibitory rates of proliferation of the OCCM-30 cells in various groups at different time points (24, 48, and 72 h); ALP (alkaline phosphatase) assay was performed to detect the cell differentiation and mineralization; real-time quantitative PCR was used to determine the expression levels of osteopontin(OPN) mRNA during 24 h; the expression levels of OPN protein at 3 and 5 d after administration were detected by Western blotting method. Results:There were no significant differences in the inhibitory rates of proliferation of OCCM-30 cells between control group and different concentrations of ursolic acid groups(P>0.05).Compared with control group, the ALP activities in the OCCM-30 cells in 2.500 μmol·L^-1 ursolic acid group at 3, 5, and 7 d after administration were significantly increased(P<0.05).Compared with control group,the expression levels of OPN mRNA and protein in the OCCM -30 cells in different concentrations of ursolic acid groups were significantly increased (P<0.01). Conclusion:Appropriate concentration of ursolic acid has no effect on the inhibitory rate of proliferation of OCCM-30 cells, but it can promote the differentiation of OCCM-30 cells and up-regulate the expression levels of OPN mRNA and protein.

Key words: ursolic acid, cementoblast, cell proliferation, cell differentiation

中图分类号:

R783.5

李梦红, 姜欢, 王六一, 冯旭, 刘楠, 胡敏. 熊果酸对小鼠成牙骨质细胞系OCCM-30细胞分化的促进作用[J]. 吉林大学学报(医学版), 2019, 45(05): 986-991.

LI Menghong, JIANG Huan, WANG Liuyi, FENG Xu, LIU Nan, HU Min. Promotion effect of ursolic acid on differentiation of mouse cementoblast cell line OCCM-30 cells[J]. Journal of Jilin University(Medicine Edition), 2019, 45(05): 986-991.

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