辛伐他汀对BMSCs成骨分化早期β-catenin及Runx2表达的影响  

Effect of simvastatin on the expression of β-catenin and Runx2 at the early stage of the osteoblast differentiation by rat bone marrow stromal cells

作　　者：贾忠宝 田发明[2] 崔纳 张柳[4] 

机构地区：[1]北京市丰台南苑医院骨外科,北京100076 [2]河北联合大学医学实验中心,河北唐山063000 [3]北京市长庚医院妇产科,北京100045 [4]河北联合大学附属医院骨外科,河北唐山063000

出　　处：《中国骨质疏松杂志》2013年第12期1237-1240,1306,共5页Chinese Journal of Osteoporosis

基　　金：河北省自然科学基金资助项目(C2006000580)

摘　　要：目的观察辛伐他汀体外给药对大鼠骨髓基质干细胞向成骨细胞分化早期β-catenin、Runx2表达的影响,探讨辛伐他汀在此过程中的作用及其作用机制。方法 20只4周龄雌性SD大鼠应用颈椎脱位法处死,无菌条件下取双侧股骨和胫骨骨髓细胞向成骨方向诱导培养。取相同第二代细胞随机分为实验组和对照组,实验组加入10-7mol/L的辛伐他汀(Simvastatin,SIM)(SIM溶于DMSO后制成母液再用完全培养基进行稀释),对照组加入含有等量DMSO的完全培养基。两组细胞在加入SIM 12 h和36 h后分别提取蛋白质和RNA,采用Western Blot法检测两组细胞β-catenin、Runx2蛋白的表达。采用实时定量-PCR法检测β-catenin、Runx2mRNA的表达。结果 (1)Western Blot结果:辛伐他汀干预12h后β-catenin和Runx2蛋白表达水平实验组与对照组相比无显著增高,两组间的差异均无统计学意义(P>0.05);辛伐他汀干预36h后实验组与对照组相比β-catenin和Runx2蛋白表达水平显著增高,两组间的差异均有统计学意义(P<0.05)。(2)实时定量-PCR结果:辛伐他汀干预12 h和36 h后β-catenin mRNA的表达水平实验组与对照组相比表达增高,两组间的差异有统计学意义(P<0.05);辛伐他汀干预12 h和36 h后Runx2 mRNA的表达水平实验组与对照组相比表达有一定程度的增高,但两组间的差异无统计学意义(P>0.05)。结论辛伐他汀体外给药对大鼠骨髓基质干细胞向成骨细胞分化早期有一定作用,这涉及到Wnt信号通路的活化,但是在不同的时间点所起作用有所差别。Objective To observe the effect of simvastatin （SIM） on the expression of β-catenin and Runx2 at the early stage of osteoblast differentiation of rat bone marrow stromal sells （BMSCs） in vitro, and to investigate the role of SIM in this process and the possible mechanism. Methods Twenty 4-week female Sprague-Dawley rats were executed by cervical dislocation. BMSCs in the bilateral femur and the tibia were collected under sterile conditions, and the cells were cultured in vitro to differentiate into osteoblasts. The 2nd-passage cells were divided into 2 groups： experimental group and control groups. Cells in experimental group were cultured with complete medium additioned with 10^-7 mol/L SIM （SIM was dissolved in DMSO first, then diluted with complete medium） , while cells in control group were cultured with complete medium additioned alone with same volume of DMSO. After 12h and 36h, the protein and RNA in 2 groups were extracted, respectively. The expression of β-catenin and Runx2 was detected using Western blotting. The mRNA expression of β-catenin and Runx2 was detected using real-time RT-PCR. Results The results of Western blotting revealed that, after 12h intervention of SIM, the expression of β-catenin and Runx2 in experimental group was not significantly higher than that in control group （P 〉 0.05）. But after 36h intervention SIM, the expression of ^-catenin and Runx2 in experimental group was significantly higher than that in control group （ P 〈 0. 05 ）. The results of real-time RT-PCR revealed that, after 12 h and 36h intervention of SIM, the mRNA expression of β-catenin in experimental group was significantly higher than that in control group （ P 〈 0.05）. After 12 h and 36h intervention of SIM, the mRNA expression of Runx2 in experimental group was higher than that in control group, but the difference was not significant （P 〉 0.05）. Conclusion SIM has some effect on the osteoblast differentiation of rat BMSCs at the early stage in vitro, which may involve the

关 键 词：辛伐他汀 骨髓基质干细胞 Β-CATENIN RUNX2 

分 类 号：R364.3[医药卫生—病理学]