HDAC2转染对骨髓干细胞成骨细胞分化的影响  

Effects of HDAC2 transfection on differentiation of bone marrow mesenchymal stem cells into osteoblasts

作　　者：李晔[1] 孙仁义[1] 吕明[1] 王鹏云[1] LI Ye;SUN Ren-yi;LV Ming;WANG Peng-yun(Department of Traumatology and Orthopaedicsf Central Hospital of Zibo City,Zibo 255036,China)

机构地区：[1]淄博市中心医院创伤骨科,山东淄博255036

出　　处：《中国矫形外科杂志》2021年第12期1119-1123,共5页Orthopedic Journal of China

基　　金：山东省医药卫生科技发展计划项目(编号:2017WS161)。

摘　　要：[目的]探讨组蛋白去乙酰化酶2(histone deacetylase 2,HDAC2)转染对骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs)向成骨细胞分化的影响。[方法]HDAC2-siRNA重组序列、阴性随机对照序列分别转入大鼠BMSCs,命名为HDAC2-si组和空载组,未做干预细胞为空白组,检测转染后各组HDAC2 mRNA表达量;诱导BMSCs成骨分化后,检测ALP活性及BGP分泌量、茜素红染色,Western blot法检测HDAC2、SHH、IHH、Ptch1、Glis1蛋白表达量。[结果]转染后HDAC2-si组HDAC2 mRNA相对表达量低于空白组和空载组(P<0.05)。茜素红染色显示,诱导前3组均无红色矿化结节,诱导后空白组和空载组出现红色矿化结节,HDAC2-si组矿化结节更多。与空白组和空载组比较,HDAC2-si组成骨诱导后HDAC2蛋白相对表达量较低,ALP活性、BGP分泌量及SHH、Ptch1、Glis1蛋白相对表达量均较高(P<0.05),IHH蛋白相对表达量组间比较,差异无统计学意义(P>0.05)。[结论]下调HDAC2表达可促进BMSCs向成骨细胞分化,机制可能是通过激活Hedgehog信号通路相关分子发挥促向成骨分化的作用。[Objective]To investigate the effect of histone deacetylase 2(HDAC2)on the differentiation of bone marrow mesenchymal stem cells(BMSCs)into osteoblasts.[Methods]The HDAC2-siRNA recombination sequence and the negative random control sequence were transferred into rat BMSCs,which were named HDAC2-si group and no-load group respectively,whereas the cells without intervention were termed as the blank group.The HDAC2 mRNA expression in each group was detected after transfection.After the osteogenic differentiation of BMSCs was induced,the ALP activity and BGP secretion were detected,the alizarin red staining was used to observe mineralization,in addition,western blot assay were conducted to determine the protein expression of HDAC2,SHH,IHH,Ptch1 and Glis1.[Results]After transfection,the relative expression of HDAC2 mRNA in HDAC2-si group was lower than that of the blank group and no-load group(P<0.05).As results of Alizarin red staining,red mineralized nodules were not seen in the three groups before induction,whereas appeared in the blank group and no-load group after induction,by contrast,more mineralized nodules were found in the HDAC2-si group.Compared with the blank group and the no-load group,the relative expression of HDAC2 protein in HDAC2-si group was lower after osteogenic induction,and the relative expression of ALP activity,BGP secretion and SHH,Ptch1,Glis1 protein were higher(P<0.05).There was no significant difference in the relative expression of IHH protein between groups(P>0.05).[Conclusion]Down-regulating the expression of HDAC2 can promote the differentiation of BMSCs into osteoblasts,which may play a role in promoting osteogenic differentiation by activating Hedgehog signaling pathway related molecules.

关 键 词：骨髓间充质干细胞 成骨细胞 分化 组蛋白去乙酰化酶2 

分 类 号：R681[医药卫生—骨科学]