辛伐他汀对幼鼠骨小梁中骨形成相关因子表达以及BMSCs成骨分化的影响  

EFFECTS OF SIMVASTATIN ON BONE FORMATION RELATIVE FACTORS OF TRABECULAR BONE AND OSTEOGENIC DIFFERENTIATION OF BONE MARROW MESENCHYMAL STEM CELLS IN YOUNG RATS

作　　者：张柳[1] 刘晓宁[1] 田发明[2] 张辉[1] 韩大成[1] 

机构地区：[1]华北煤炭医学院附属医院骨外科河北唐山,063000 [2]河北医科大学研究生院

出　　处：《中国修复重建外科杂志》2011年第2期155-159,共5页Chinese Journal of Reparative and Reconstructive Surgery

基　　金：河北省自然科学基金资助项目(C2006000580)~~

摘　　要：目的既往研究表明辛伐他汀具有促成骨作用,现探讨辛伐他汀对幼鼠骨小梁中骨形成相关因子表达以及BMSCs成骨分化的影响。方法 40只1周龄雄性SD大鼠,体重17.5～19.5 g,随机分为2组,每组20只。实验组于大鼠颈部皮下注射辛伐他汀注射液[5 mg/(kg.d)],对照组同法注射等剂量生理盐水。于注射后1、3周,两组分别脱颈处死10只大鼠,采用免疫组织化学染色检测胫骨上端骨小梁BMP-2、基质金属蛋白酶13(matrix metalloproteinase 13,MMP-13)、VEGF的表达情况。注射后1、3周,两组分别取大鼠双侧股骨,采用全骨髓培养法体外分离培养BMSCs,取第1代BMSCs成骨诱导培养,培养后14 d行ALP染色,培养后21 d采用实时荧光定量PCR检测BMP-2、Runx2、Osterix、Msx2、Dlx3、Dlx5 mRNA的表达,培养后28 d行von Kossa染色。结果免疫组织化学染色示注射后1、3周两组胫骨上端骨小梁BMP-2、MMP-13、VEGF表达,差异均无统计学意义(P>0.05)。成骨诱导培养14 d,注射后1、3周两组ALP染色阳性细胞百分比差异均无统计学意义(P>0.05);成骨诱导培养21 d,实时荧光定量PCR检测示注射后1、3周两组BMP-2、Runx2、Osterix、Dlx3、Dlx5、Msx2 mRNA表达水平比较差异均无统计学意义(P>0.05);成骨诱导培养28 d,注射后1、3周两组钙结节单位面积比差异均无统计学意义(P>0.05)。结论皮下注射辛伐他汀5 mg/(kg.d)后1、3周对幼鼠骨小梁中骨形成相关因子表达以及BMSCs成骨分化无明显影响。Objective Simvastatin has been reported to be effective on stimulation of bone formation.To investigate the effects of simvastatin on bone formation relative factors of proximal tibia trabecular bone and on osteogenic differentiation of bone marrow mesenchymal stem cells（BMSCs）.Methods Fourty 1-week-old male Sprague Dawley rats were divided randomly into 2 groups,20 rats per group.Rats in experimental group received subcutaneous injection of simvastatin [（5 mg/（kgo d）],and the rats in control group received injection of normal saline at the same dose.The expressions of bone morphogenetic protein 2（BMP-2）,matrix metalloproteinase 13（MMP-13）,and vascular endothelial growth factor（VEGF） of trabecular bone were analyzed in the tibia by immunohistochemical staining at 1 and 3 weeks after injection.BMSCs from the rat femur at 1 and 3 weeks after injection were cultured under condition of osteogenic induction.ALP staining was performed on the 14th day after culture;real-time fluorescent quantitative PCR was used to detect the mRNA expressions of BMP-2,Runx2,Osterix,Msx2,Dlx3,and Dlx5 on the 21st day after culture;and von Kossa staining was performed on the 28th day after culture.Results There was no significant difference in the expressions of BMP-2,MMP-13,and VEGF between the experimental group and control group at 1 and 3 weeks after injection（P 〉 0.05）.There was no significant difference in the percentages of ALP positively-stained cells between the experimental group and the control group on the 14th day after culture（P 〉 0.05）.The mRNA expressions of BMP-2,Runx2,Osterix,Msx2,Dlx3,and Dlx5 in osteogenic differentiation-induced BMSCs had also no significant difference between the experimental group and the control group at 1 and 3 weeks after culture（P 〉 0.05）.No significant difference in biomineralization was found between the experimental group and control group at 1 and 3 weeks after culture（P 〉 0.05）.Conclusion Subcutaneous injection of simvastatin [（5 mg/（kgod）

关 键 词：辛伐他汀 BMSCS 成骨分化 骨形成相关因子 大鼠 

分 类 号：R965[医药卫生—药理学]