不同浓度熊果酸对破骨细胞增殖分化的作用及其意义

发布时间：2018-04-11 12:08

本文选题：熊果酸 + 破骨细胞　；参考：《吉林大学》2017年硕士论文

【摘要】：目的:研究熊果酸(UA)对破骨样细胞(OC)增殖分化的影响,探讨其在正畸牙根吸收过程中的作用及与OC之间的关系。方法:对小鼠单核/巨噬细胞系RAW264.7细胞进行破骨样细胞诱导,采用抗酒石酸酸性磷酸酶(TRAP)染色法及骨吸收陷窝实验鉴定诱导是否成功。采用CCK-8法选取对RAW264.7细胞无细胞生物毒性的适宜浓度的UA,观察其对RAW264.7增殖分化影响。实验组在培养液中加入梯度浓度1.0、2.5、5.0、10.0、20.0和40.0μmol·L~（-1） UA,对照组不加UA。采用RT-PCR方法检测适宜浓度UA(5.0μmol·L~（-1）)与OC培养第5d时OC标志性基因TRAP、NFATc1水平表达,观察其对OC分化的影响。结果:TRAP染色及骨吸收陷窝观察,诱导培养RAW264.7细胞5d后,可见TRAP染色阳性细胞;扫描电镜下吸收陷窝呈圆形、椭圆形等,底壁较粗糙,边界较清晰。表明RAW264.7细胞成功向破骨样细胞分化。CCK-8检测表明,高浓度UA(10.0μmol·L~（-1）)显著抑制OC增殖;适宜浓度UA(5.0μmol·L~（-1）),既在生物安全浓度范围内,又抑制OC增殖;低浓度UA(2.5μmol·L~（-1）)无作用;且第5d时,中浓度UA可显著抑制OC标志性基因的表达水平,说明UA可抑制OC分化。结论:UA与OC增殖分化有关联,其在适宜浓度(5.0μmol·L~（-1）)下对OC起抑制作用,且呈时间依赖性。
[Abstract]:Aim: to study the effect of ursolic acid (UAA) on the proliferation and differentiation of osteoclasts (OCs), and to explore the role of UAA in the root resorption of orthodontic teeth and its relationship with OC.Methods: osteoclast-like cells were induced from mouse mononuclear / macrophage RAW264.7 cells. The results of tartrate-resistant acid phosphatase staining and bone resorption trap test were used to determine whether the induction was successful or not.CCK-8 method was used to select the appropriate concentration of RAW264.7 cells without cytotoxicity and to observe the effect of UAU on the proliferation and differentiation of RAW264.7 cells.In the experimental group, the gradient concentration of 1.0 渭 mol / L and 40.0 渭 mol / L ~ (-1) UAA was added to the culture medium, while the control group was not added with UA.The RT-PCR method was used to detect the expression of OC signature gene TRAPN ATC1 at the 5th day of culture with the appropriate concentration of UA(5.0 渭 mol L ~ (-1) and OC, and to observe its effect on the differentiation of OC.Results the positive cells of TRAP staining were found in RAW264.7 cells after 5 days of induction and culture, and the absorptive lacunae were round, elliptical, rough in bottom wall and clear in boundary under scanning electron microscope.The medium concentration of UA could significantly inhibit the expression of OC signature gene, indicating that UA could inhibit the differentiation of OC.Conclusion there is a correlation between the proliferation and differentiation of OC at the appropriate concentration of 5.0 渭 mol / L ~ (-1), and it has a time-dependent inhibitory effect on OC.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R783.5

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