摘要：本次实验的主要目的是为了探究辛伐他汀（1×10-7 mol/L）对成骨细胞早期粘附与分化的影响。本研究首先选取SD大鼠的乳鼠（出生1-3天内）的头盖骨组织进行体外培养，待细胞爬出长满传至第三代时，取其进行分组实验，实验组给予含辛伐他汀（1×10-7 mol/L）的完全培养液对其干预，而对照组给予含等量溶剂无水乙醇的完全培养液进行干预，待培养24小时后观察其细胞的形态，分别培养4和7天后对不同基底细胞的碱性磷酸酶活性定量测定。本研究对粘附性的影响主要通过观察纽蛋白和肌动蛋白表达的情况，而对成骨分化的影响主要通过测定与底物对硝基苯酚（PNP）的反应情况来反映碱性磷酸酶酶活性大小，结果表明辛伐他汀（1×10-7 mol/L）对成骨细胞早期粘附无明显作用而对成骨细胞的形态和分化有明显的促进作用。69049

毕业论文关键词：辛伐他汀、成骨细胞、粘附、分化、碱性磷酸酶活性

Effect of simvastatin on early adhesion and differentiation of osteoblasts

Abstract: The main purpose of this study was to explore the effect of simvastatin(1 x 10-7 mol / L) on the early adhesion and differentiation of osteoblasts. This study firstly isolated the skull bone tissue of newborn SD rats (postnatal 1-3 days) and cultured in vitro, the cells were passaged to the third generation and then above respective experiments were performed, including the experimental group give containing simvastatin (1 x 10-7 mol / L) in complete culture medium and the contrast group containing the same amount of solvent ethanol in the complete culture medium. when cells were cultured for 24 hours, the morphological observation was conducted, when they were cultured for 4 and 7 days, the alkaline phosphatase activities were quantitatively determined. In this study, the expression of vinculin and actin were marked as the adhesion, and the measurement of substrate p-nitrophenol (PNP) response was used to reflect the activity of the alkaline phosphatase enzyme. The results showed that simvastatin（1×10-7 mol/L）had no obvious effect on early adhesion of bone cells, bone cell morphology and differentiation were promoted by simvastatin .

key word：Simvastatin, osteoblast, adhesion, differentiation, alkaline phosphatase activity

目录

引言 1

1 材料与方法 2

1.1供试材料 2

1.1.1动物 2

1.2 药品与试剂 2

1.3 方法 2

        1.3.1DMEM高糖培养基的配置 2      

1.3.2原代培养 2

        1.3.3成骨细胞ALP染色的鉴定 3

1.3.4实验分组 3

1.3.5细胞形态观察 3

        1.3.6细胞内蛋白总量测定 4

1.3.7碱性磷酸酶活性检测 5

2 结果与分析 5

    2.1 成骨细胞的形态及鉴定 5

2.2 辛伐他汀对成骨细胞早期粘附的影响 6

2.3 辛伐他汀对成骨细胞分化的影响 7

3 讨论 8

4 致谢 9

参考文献 10