骨髓基质干细胞成骨分化过程中Wnt、骨形态发生蛋白2信号通路相关因子与辛伐他汀的作用  

Effects of simvastatin on Wnt and bone morphogenetic protein 2 signaling pathway during osteoblast differentiation of bone marrow stromal cells

作　　者：靳云乔[1] 张柳[1] 田发明[1] 张磊[1] 程潭[1] 刘晓宁[1] 

机构地区：[1]河北联合大学附属医院骨外科,河北省唐山市063000

出　　处：《中国组织工程研究与临床康复》2011年第36期6732-6736,共5页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：河北省自然科学基金资助项目(C2006000580)~~

摘　　要：背景:辛伐他汀可促进体外培养的人或鼠骨髓基质干细胞向成骨细胞分化,但作用机制尚不清楚。目的:观察辛伐他汀对大鼠骨髓基质干细胞向成骨细胞分化过程中Wnt与骨形态发生蛋白2信号途径中相关因子表达的影响。方法:取6周龄雌性SD大鼠双侧股骨、胫骨全骨髓进行体外成骨细胞诱导培养。实验分为对照组及SIM组。SIM组加入浓度为10-7mol/L辛伐他汀,对照组加入等量无水乙醇和PBS。培养14d,行碱性磷酸酶染色,28d时,行vonKossa染色观察细胞外基质矿化情况;培养14,21d,免疫荧光细胞化学染色观察成骨细胞中β-catenin,Smad1/5,Cbfa1的表达及分布。结果与结论:大鼠骨髓基质干细胞经体外诱导后可分化为具有碱性磷酸酶活性和矿化细胞外基质能力的成熟成骨细胞。辛伐他汀可显著上调骨髓基质干细胞成骨分化过程中碱性磷酸酶的表达。同时,与对照组比较,SIM组β-catenin,Smad1/5,Cbfa1表达明显增多(P<0.05),且呈现明显的核内聚集趋势。说明辛伐他汀促进骨髓基质干细胞向成骨细胞分化的作用可能与调控Wnt与骨形态发生蛋白2信号通路中相关因子的表达及细胞内分布有关。BACKGROUND：Simvastatin can promote osteoblast differentiation of human or rat bone marrow stromal stem cells cultured in vitro.But the mechanism remains unclear.OBJECTIVE：To investigate the effects of simvastain on expression of factors related to Wnt and bone morphogenetic protein 2（BMP-2） signaling pathway during osteoblast differentiation of bone marrow stromal cells.METHODS：Bilateral femur and tibia bone marrow was harvested from 6-week-old female Sprague-Dawley rats for in vitro culture of osteoblasts.In the SIM group,10-7 mol/L simvastain was added,and in the control group,the same amount of dehydrated alcohol and PBS was added.After 14 days of culture,alkaline phosphatase staining was performed.At 28 days,von Kossa staining was applied to observe osteoblastic differentiation and mineralization.At 14 and 21 days,immunofluorescent cytochemical staining was used to detect β-catenin,Smad1/5,Cbfa1 expression and distribution in osteoblasts.RESULTS AND CONCLUSION：Cells in each group were differentiated into osteoblast lineage,as shown by alkaline phosphatase activity and extracellular matrix mineralization.Simvastatin can significantly upregulate alkaline phosphatase expression during the osteoblast differentiation of bone marrow stromal cells.The expression of β-catenin,Smad1/5,and Cbfa1 was significantly increased in the SIM group than in the control group.The above-mentioned expression in the SIM group accumulated in the nuclear more than in the cytoplasm.These results showed that the mechanism by which simvastatin promotes osteoblast differentiation of bone marrow stromal stem cells may be partially related to expression and distribution of factors regulating Wnt and BMP-2 signaling pathway

关 键 词：骨髓基质干细胞 成骨细胞 WNT信号通路 骨形态发生蛋白2信号通路 辛伐他汀 

分 类 号：R394.2[医药卫生—医学遗传学]