›› 2014, Vol. 5 ›› Issue (3): 113-117.

• 论著 •    下一篇

组蛋白去乙酰化酶8对大鼠骨髓间充质干细胞成骨分化的影响

周华,傅瑜,戈杰,周培培,江宏兵   

  1. 江苏省南京医科大学附属口腔医院口腔颌面外科
  • 收稿日期:2014-04-09 修回日期:2014-08-04 出版日期:2014-09-25 发布日期:2014-09-24
  • 通讯作者: 江宏兵 E-mail:jhbtooth@163.com
  • 基金资助:
    国家自然科学基金;江苏高校优势学科建设工程资助项目

Effect of histone deacetylase 8 on osteoblast differentiation of rat bone marrow mesenchymal stem cells

  • Received:2014-04-09 Revised:2014-08-04 Online:2014-09-25 Published:2014-09-24

摘要: 目的:探讨组蛋白去乙酰化酶8(histone deacetylase 8,HDAC8)对大鼠骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs)成骨分化的影响。方法:通过全骨髓贴壁法体外分离培养大鼠BMSCs,转染HDAC8过表达慢病毒载体,并于矿化诱导液中培养7 d后,实时荧光定量PCR、Western blot检测BMSCs成骨分化能力的变化;矿化诱导14 d后,茜素红钙结节染色检测BMSCs矿化能力的变化。组蛋白去乙酰化酶抑制剂——曲古抑菌素A(trichostatin A,TSA)刺激转染HDAC8过表达慢病毒载体的BMSCs,于矿化诱导液中培养7 d后,实时荧光定量PCR及Western blot检测TSA刺激前后过表达HDAC8的BMSCs成骨分化能力的变化。结果:HDAC8过表达组经矿化诱导7 d后,成骨分化相关基因mRNA、蛋白表达水平均低于空白对照组;经14 d矿化诱导后,HDAC8过表达组茜素红钙结节的着色程度及范围低于空白对照组。TSA刺激的HDAC8过表达组矿化诱导7 d后,成骨分化相关基因mRNA及蛋白的表达均高于未加刺激组(P<0.05)。结论:HDAC8对大鼠BMSCs成骨分化具有抑制作用。

Abstract: Objective:To evaluate the effect of histone deacetylase 8 (HDAC8) on osteoblast differentiation of rat bone marrow mesenchymal stem cells (BMSCs). Methods:Rat BMSCs were isolated and cultured by the method of whole bone marrow adherent and transfected with HDAC8 overexpression lentiviral vector. Real-time PCR, Western blot were applied to estimate the change of osteogenic capacity after 7 days of osteogenic induction and Alizarin red stain was used to estimate the mineralization capacity after 14 days of osteogenic induction. Trichostatin A (TSA), one kind of histone deacetylase inhibitor, acted on BMSCs with HDAC8 overexpression and the change of osteogenic capacity of cells after 7 days of osteogenic induction was estimated by real-time PCR and Western blot. Results:For BMSCs with HDAC8 overexpression, the expression of osteogenesis-related genes at mRNA and protein levels were all lower than those in control group after 7 days of ostogenic induction and the capacity to form calcified nodules was lower than that of the control group after 14 days of ostogenic induction. The osteogenesis-related genes expression of BMSCs with HDAC8 overexpression which was treated with TSA were all higher than those in untreated group after 7 days of ostogenic induction (P<0.05). Conclusions:HDAC8 could suppress osteogenic differentiation of rat BMSCs.

中图分类号: