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Satb2过表达慢病毒载体构建及其对骨髓基质细胞的转染及表达
  • ISSN号:1674-8603
  • 期刊名称:口腔生物医学
  • 时间:2013.6.6
  • 页码:61-64
  • 分类:R780.2[医药卫生—口腔医学;医药卫生—临床医学]
  • 作者机构:[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
  • 相关基金: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)
  • 相关项目:表观遗传修饰不同胚源MSCs促进颌骨成骨修复及相关机制研究
中文摘要:

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。

英文摘要:

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.

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期刊信息
  • 《口腔生物医学》
  • 主管单位:江苏省教育厅
  • 主办单位:南京医科大学
  • 主编:陈宁
  • 地址:江苏省南京市汉中路136号南京医科大学附属口腔医院911室
  • 邮编:210029
  • 邮箱:kqswyx@126.com
  • 电话:025-85031861
  • 国际标准刊号:ISSN:1674-8603
  • 国内统一刊号:ISSN:32-1813/R
  • 邮发代号:28-64
  • 获奖情况:
  • 国内外数据库收录:
  • 被引量:461