摘要
目的研究在周期性牵张力介导下,骨形态发生蛋白9(bone morphogenetic proteins 9,BMP9)能否激活PI3K/AKT信号通路调控人牙周膜成纤维细胞(human periodontal ligament fibroblasts,hPDLFs)成骨分化。方法利用FlexCell系统对体外培养的hPDLFs加载正弦波、形变率10%、频率0.5Hz的周期性牵张力,免疫荧光法检测细胞骨架蛋白的表达及分布,qPCR检测RUNX2、OCN、OPN、OSX mRNA的表达情况,免疫印迹法检测成骨标志蛋白OCN、OPN的表达,加入PI3K信号抑制剂LY294002后AKT、P-AKT以及OCN、OPN的变化。结果与对照组比较,6 h、12 h组的细胞骨架蛋白表达增多且呈受力方向分布,OCN、OPN表达上调,差异有统计学意义(P<0.05),qPCR检测mRNA的表达趋势与蛋白检测结果一致。结论周期性牵张力介导BMP9可以通过PI3K/AKT信号通路调控hPDLFs成骨分化。
Objective To investigate whether bone morphogenetic proteins 9(BMP9)can activate PI3 K/AKT signaling pathway to regulate osteogenic differentiation of human periodontal ligament fibroblasts(hPDLFs)under cyclic tension force.Methods The cultured in vitro hPDLFS were loaded with sine wave,deformation rate 10%,frequency 0.5 Hz periodic tension by FlexCell system.Distribution and changes of cytoskeletal protein expression were observed by immunofluorescence method.Changes of mRNA expression of RUNX2,OCN,OPN and OSX were observed by qPCR.Cytoskeletal protein OCN,OPN expressions and changes of AKT,P-AKT,OCN,OPN after adding P13 K signal inhibitor LY294002 were detected by Western blot.Results Compared with control group,there were a significant increase of the expression of cytoskeletion-associated protein,up-regulation of the expression of osteogenic markers in 6 h and 12 h groups(P<0.05).The trend of mRNA expression was consistent with the result of protein detection.Conclusions Cyclic tension force-mediated BMP9 can regulate osteogenic differentiation of hPDLFs through PI3 K/AKT signaling pathway.
作者
张超
胡静
张君怡
魏克元
孙海豹
黄克强
ZHANG Chao;HU Jing;ZHANG Jun-yi;WEI Ke-yuari;SUN Hai-bao;HUANG Ke-qiang(The Second Affiliated Hospital of Jinzhou Medical University;Jinzhou Medical University Life Science Research Institute;Jinzhou Medical University School of Rehabilitation and Sports,Jinzhou 121000,Liaoning Province;Affiliated Hospital of Chifeng University,Chifeng 024000,Inner Mongolia;Shaoxing Stomatological Hospital,Shaoxing 312000,Zhejiang Province,China)
出处
《中国临床解剖学杂志》
CSCD
北大核心
2019年第6期656-661,共6页
Chinese Journal of Clinical Anatomy
基金
锦州医科大学校长基金（AH2015002
