摘要：目的 通过体外给药的方式用辛伐他汀直接干预大鼠骨髓间充质干细胞（bone marrow stromal cells， BMSCs），从而研究辛伐他汀对骨髓间充质干细胞早期粘附与成骨分化的影响。方法 从7-8周龄SD大鼠股骨和胫骨中分离得到原代BMSCs后进行体外培养，取第三代细胞接种于实验组（含1×10-7M辛伐他汀的完全培养基）和对照组中（等量的完全培养基）。培养1天后，通过免疫荧光染色观察细胞形态；分别培养1天和7天后，对培养细胞进行碱性磷酸酶（alkaline phosphatase，ALP）定量测定。结果 通过染色观察可知1×10-7M辛伐他汀促进了BMSCs的早期铺展，对它的早期粘附并没有显著的影响。实验组较对照组而言，其碱性磷酸酶活性更高，结果表明1×10-7M辛伐他汀促进了BMSCs早期的成骨分化。69048

毕业论文关键词：辛伐他汀、大鼠骨髓间充质干细胞、粘附、碱性磷酸酶、成骨分化

Effect of simvastatin on early adhesion and osteogenic differentiation of bone marrow mesenchymal stem cells

Abstract: Objective : To study the effect of simvastatin on the early stage of adhesion and osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) by in vitro administration of system. Methods : 7 the BMSCs were isolated and cultured in vitro from the femur and tibia SD rats (7-8 weeks old), and  the third generations of cells were used  in the experimental group (1 x 10-7M simvastatin) and control group (equivalent total medium). After 1 days of culture, the cell morphologies were observed.After cultured for 1 days and 7 days respectively, and The alkaline phosphatase (ALP) activity was determined by immunofluorescence staining. Results : In contrast with the control group, the alkaline phosphatase activity was higher in the experimental group.The results showed that simvastatin had no significant effect on the early adhesion of BMSCs, however, promoted the early osteogenic differentiation of BMSCs.

Key words: Simvastatin; rat bone marrow mesenchymal stem cells; adhesion; alkaline phosphatase; osteogenic differentiation

目录

前言 4

1.实验材料与方法 5

1.1 实验动物 5

1.2 实验药品及试剂 5

1.3 实验方法 5

1.3.1 BMSC的提取及培养 5

1.3.2 实验分组及进行药物干预 6

1.3.3 细胞的染色观察 6

1.3.4 细胞内的总蛋白含量测定 7

1.3.5 碱性磷酸酶活性的检测 7

2.实验结果 7

2.1 BMSC的生长状态及形态观察 7

2.2辛伐他汀对BMSCs粘附的影响 9

2.3辛伐他汀对BMSCs早期分化的影响 10

3.讨论 11

4.致谢 12

参考文献 12

前言

随着我国经济飞速发展，中国人口老龄化趋势日益严重，骨质疏松成为了一种常见疾病。骨质疏松是指骨量缓慢流失，骨组织的微观结构发生改变，骨质变脆弱，导致更加容易引发骨折和骨痛等一系列并发症。当前医学，用于治疗骨质疏松的主要药物的药理作用是减缓骨质流失，然而少有药物是用来直接促进成骨的。因此，当前进行的针对促进成骨作用药物的研究对骨质疏松症的治疗有着十分重要的帮助。