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目的:研究辛伐他汀对原代培养大鼠颅骨成骨细胞增殖、分化及矿化功能的影响。方法:组织块法分离培养大鼠颅骨成骨细胞,分别在含不同浓度辛伐他汀(1.0μmol/L、0.5μmol/L、0.25μmol/L、0.125μmol/L)培养液环境下培养成骨细胞,采用MTT法测定细胞增殖情况;检测细胞碱性磷酸酶(ALP)活性以及培养液中骨钙素(OC)含量;Von Kossa染色观察成骨细胞矿化结节,用Image-Pro Plus(IPP6.0)软件及其相关系统完成图像采集及分析计算成骨细胞矿化面积的百分比,通过矿化结节面积百分比计算检测细胞的矿化能力。采用SPSS13.0对数据进行单因素方差分析。结果:辛伐他汀抑制成骨细胞的增殖,呈剂量依赖性。各浓度组的ALP活性均增加,以0.250μmol/L浓度组的作用效应最为显著;骨钙素水平增加,与对照组比较均有统计学意义;辛伐他汀能使成骨细胞矿化面积百分比显著增加,差异有统计学意义(P<0.05)。结论:辛伐他汀抑制体外培养大鼠颅骨成骨细胞的增殖,但却促进成骨细胞的分化及矿化,发挥促进骨形成的作用。
Objective: To study the effect of simvastatin on proliferation, differentiation and mineralization of primary cultured rat skull osteoblasts. METHODS: Rat calvarial osteoblasts were isolated and cultured by tissue block method. Osteoblasts were cultured in the culture medium containing different concentrations of simvastatin (1.0μmol / L, 0.5μmol / L, 0.25μmol / L, 0.125μmol / L) The cell proliferation was assayed by MTT assay. The activity of alkaline phosphatase (ALP) and the content of osteocalcin (OC) in the culture medium were detected. Von Kossa staining was used to observe the mineralized nodules of osteoblasts. IPP6.0) software and related systems to complete the image acquisition and analysis of osteoblast mineralization area percentage calculated by the percentage of mineralized nodule area calculation of cell mineralization ability. Data were analyzed by one-way ANOVA using SPSS 13.0. Results: Simvastatin inhibited osteoblast proliferation in a dose-dependent manner. ALP activity in all concentration groups increased, and the effect of 0.250μmol / L group was the most significant; osteocalcin levels increased, compared with the control group were statistically significant; simvastatin can osteoblast mineralization area percentage Significantly increased, the difference was statistically significant (P <0.05). Conclusion: Simvastatin can inhibit the proliferation of rat calvarial osteoblasts in vitro, but promote the differentiation and mineralization of osteoblasts and promote bone formation.