中国临床解剖学杂志 ›› 2015, Vol. 33 ›› Issue (4): 434-438.doi: 10.13418/j.issn.1001-165x.2015.04.013

• 实验研究 • 上一篇    下一篇

辛伐他汀对BMSCs成骨分化过程中TGF-β1/ smad信号通路的影响

桑遥彩1, 包翠芬1, 武蕾2, 张苗苗1, 王艳1, 秦书俭1   

  1. 1.辽宁医学院人体解剖与组织胚胎学教研室,辽宁 锦州 121000; 2.日照市东港区人民医院口腔科,山东 日照 276800
  • 收稿日期:2014-12-11 出版日期:2015-07-25 发布日期:2015-08-14
  • 通讯作者: 秦书俭,教授,博士生导师,E-mail:mianyizuhua @aliyun.com
  • 作者简介:桑遥彩(1988-)男,山东日照人,在读硕士,研究方向:骨组织工程,Tel:15940633971, E-mail:723605766@qq.com
  • 基金资助:

     国家自然科学基金(31170930, 81202783)

Effects of simvastatin on TGF-β1/smad signaling pathway in the process of the differentiation of BMSCs

SANG Yao-cai1,   BAO Cui-fen1,   WU Lei2,   ZHANG Miao-miao1,   WANG Yan1,   QIN Shu-jian1   

  1. 1.Department of Human Anatomy and Histology and Embryology, Liaoning Medical University, Jinzhou, Liaoning 121000, China; 2.Department of Oral,  Rizhao Donggang District People's Hospital, Rizhao,Shandong 276800, China
  • Received:2014-12-11 Online:2015-07-25 Published:2015-08-14

摘要:

目的 探讨辛伐他汀(simvastatin,SIM)是否可以通过调节TGF-β1/Smad通路从而促进大鼠骨髓间充质干细胞(bone marrow mesenchymal stem cells , BMSCs)分化为成骨样细胞。 方法 复苏、培养大鼠BMSCs并诱导其向成骨样细胞分化。实验组加入不同浓度的SIM(10-8 mol/L、10-7 mol/L、10-6 mol/L),对照组不给予药物干预。加入成骨诱导液,7d后运用碱性磷酸酶染色法观察细胞成骨能力,21d后用茜素红染色检测细胞钙化水平,Western blot法检测TGF-β1、Smad2、Smad3蛋白的表达水平。  结果 应用10-8 mol/L~10-6 mol/L SIM处理细胞7d均可使细胞ALP活性显著增加,其中浓度为10-7 mol/L时效果最显著;应用10-8 mol/L~10-6mol/L SIM处理细胞21d后可使细胞钙化结节明显增多;应用10-8 mol/L~10-6 mol/L SIM处理细胞7d后细胞内TGF-β1、Smad2、Smad3蛋白水平明显增高。 结论 辛伐他汀可促进大鼠BMSCs分化为成骨细胞,其作用机制可能与上调TGF-β1/Smad信号通路中相关基因的表达水平有关。

关键词: 辛伐他汀, 骨髓间充质干细胞, 成骨细胞, 转化生长因子-&beta, 1

Abstract:

Objective    To investigate whether simvastatin(SIM) can promote differentiation of rat bone marrow mesenchymal stem cells(BMSCs ) to osteablasts by regulating TGF-β1/Smad signaling pathways.  Methods   Rat BMSCs were recovered, cultured and further induced to differentiate into osteoblasts. Different concentration of SIM(10-8 mol/L、10-7 mol/L、10-6 mol/L) was added in the experimental group, and the control group no drug intervention was given in the control group. 7 days after ostrogenic solution was added, alkaline phosphatase staining was used to observe the expression level of bone formation, Alizarin Red staining was used to observe calcification level,and Western blot was used to determine the expression of of TGF-β1,Smad2 and Smad3.   Results   Treatment with SIM at concentrations of 10-8 mol/L to 10-6 mol/L for 7 d  significantly increased the activity of ALP, and SIM at concentration of 10-7 mol/L produced the maximum effect. Exposure of the cells to SIM at concentration of 10-8 mol/L~10-6 mol/L for 21 d significantly increased mineralized nodules. Exposure of the cells to SIM at concentrations of 10-8~10-6 mol/L for 7 d markedly increased the expression of TGF-β1, Smad2 and Smad3.   Conclusion   SIM could promote the osteogenic differentiation of BMSCs,in which process the changes of the mRNA expression levels inTGF-β1/Smad signaling pathway might participate.

Key words: Simvastatin, Bone marrow mesenchymal stem cells, Osteoblasts, TGF-β1