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  

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

作　　者：周华[1,2] 傅瑜[1,2] 戈杰[1,2] 周培培[1,2] 江宏兵[1,2] 

机构地区：[1]南京医科大学口腔疾病研究江苏省重点实验室,江苏南京210029 [2]南京医科大学附属口腔医院口腔颌面外科,江苏南京210029

出　　处：《口腔生物医学》2014年第3期113-117,共5页Oral Biomedicine

基　　金：国家自然科学基金(81070810);江苏高校优势学科建设工程资助项目(2014-37);南京医科大学科技发展基金(2013NJMU059)

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。Objective：To evaluate the effect of histone deacetylase 8 （HDAC8）on osteoblast differentiation of rat bone marrow mes-enchymal 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 osteogen-ic capacity after 7 days of osteogenic induction and Alizarin red stain was used to estimate the mineralization capacity after 1 4 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 low-er 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 1 4 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.

关 键 词：组蛋白去乙酰化酶8 组蛋白去乙酰化酶抑制剂 骨髓间充质干细胞 成骨分化 大鼠 

分 类 号：R780.2[医药卫生—口腔医学]