摘要
目的:研究特异性糖原合成酶激酶-3β(GSK-3β)抑制剂Tideglusib对小鼠成骨细胞MC3T3-E1增殖和成骨相关基因表达的影响。方法:采用CCK-8法测定梯度浓度(0~2000 ng/L)Tideglusib培养1、2、3 d时MC3T3-E1细胞的增殖活性,筛选有效浓度;采用荧光显微镜观察有效浓度Tideglusib培养8 h时MC3T3-E1细胞的黏附铺展形态;采用实时定量PCR仪检测有效浓度Tideglusib培养7、14 d时MC3T3-E1细胞中Runt相关转录因子2(Runx2)、Osterix(OSX)、碱性磷酸酶(ALP)、骨桥蛋白(OPN)、骨钙素(OCN)等成骨相关基因的表达水平,Western blot法检测其Runx2、OCN的蛋白表达水平。结果:与对照组相比,10、20 ng/L浓度Tideglusib具有促进MC3T3-E1细胞的增殖和黏附作用;10、20 ng/L浓度Tideglusib可上调Runx2、OSX、OPN、OCN的基因表达水平,并增强Runx2、OCN的蛋白表达水平。结论:Tideglusib能够促进体外成骨细胞的增殖、黏附以及成骨相关基因的表达。
Objective:To explore the effects of Tideglusib,a specific chemical inhibitor of GSK-3β,on proliferation and osteogenic gene expression of mouse osteoblast-like MC3T3-E1 cells.Methods:The proliferation of MC3T3-E1 cells was detected by CCK-8 assay after culturing in gradient concentrations(0-2000 ng/L)of Tideglusib for 1,2 and 3 days,and the effective concentration of Tideglusib was screened.The adhesion and spreading morphology of MC3T3-E1 cells culturing with the effective concentration of Tideglusib for 8 h was observed by fluorescence microscope.The mRNA expression of Runx2,OSX,ALP,OPN and OCN of MC3T3-E1 cells was analyzed by real-time PCR and the protein expression of Runx2,OCN was detected by Western blot after culturing with the effective concentration of Tideglusib for 7 and 14 days.Results:Compared with the control group,Tideglusib could promote proliferation and adhesion abilities of MC3T3-E1 cells at the effective concentration of 10 and 20 ng/L.The effective concentration of Tideglusib could up-regulated the mRNA levels of Runx2,OSX,ALP,OPN and OCN and the protein levels of Runx2 and OCN.Conclusions:Tideglusib can enhance the proliferation and osteogenic gene expression activities of osteoblasts in vitro.
作者
于小宇
朱文卿
邱憬
YU Xiaoyu;ZHU Wenqing;QIU Jing(Department of Oral Implantology,Jiangsu Key Laboratory of Oral Disease,Affiliated Stomatological Hospital of Nanjing Medical University,Nanjing 210029,China)
出处
《口腔生物医学》
2019年第4期191-195,共5页
Oral Biomedicine
基金
国家自然科学基金面上项目(81870799)
江苏省重点研发计划(社会发展)项目(BE2019728)
江苏省青年医学人才项目(QNRC2016850)
南京医科大学-苏盐智创新研发基金项目(NMU-SY201806)
江苏高校优势学科建设工程资助项目(2018-87)
