rhBMP2作用下人牙周膜成纤维细胞中OCIF、ODF的表达研究

目的：本研究通过反转录聚合酶链反应来测定不同浓度的重组人骨形成蛋白2（rhBMP2）对人牙周膜成纤维细胞中OCIF、ODFmRNA在不同作用时间点表达的影响,了解rhBMP2对骨改建相关基因的调控方式。方法：原代培养人牙周膜成纤维细胞,取生长良好的第六代细胞,分别用25ng/ml、50 ng/ml及100 ng/ml的rhBMP2作用于细胞,于作用1、3、5、7天后收集细胞。反转录聚合酶链反应检测各组细胞四个时间作用点OCIF、ODF mRNA含量,PCR产物1%琼脂糖凝胶电泳后,采用Image Pro Plus5.0图像分析软件对电泳胶带亮度进行分析。结果：在rhBMP2作用初期,由于OCIF mRNA的表达快速上升,ODF mRNA的表达缓慢增高,二者的表达差异远大于正常生理水平;随作用时间的延长,由于OCIF mRNA的表达下降,ODF mRNA的表达缓慢增高,二者的表达差异逐渐缩小,在作用末期低于正常生理水平。结论：基于OCIF与ODF二者表达的相对变化,HPDLfs在rhBMP2的作用下可能对破骨细胞的形成在初期表现为较强的抑制,随着作用时间的延长,HPDLfs对破骨细胞的抑制作用逐渐转变成为促进其活化的作用。

Study of the expression variety of OCIF and ODF in human periodontal ligament fibroblasts with rhBMP2 reaction

Objective By RT-PCR to investigating the effects of rhBMPs in different concentration on the expression of OCIF、ODF in human periodontal ligament fibroblasts.To clarify the molecular mechanism of BMPs＇ regulation to the HPDLfs,which contribute to the bone rebuilding.Methods The 6th generation well-grown primary culture fibrocyte from HPDLfs were used in the experiment.The cells were subjected to different doses of rhBMP2（25ng/ml,50ng/ml and 100ng/ml） for 1,3,5,7 days in this experiment.mRNA expression of OCIF、ODF were determined by semiquantitative RT-PCR approach.electrophoresis tape brightness was analyzed by graphical analysis software Image Pro Plus5.0.Results The expression varieties between cbfα1 and OCIF in different concentration were compared.At primary stage,their expression varieties were concordance and presented positive correlation.But the variety correlation between cbfα1 and OCIF was no more definitude after OCIF reaching peak value with the action time prolonging.Conclusion Based on the varieties of expression between OCIF and ODF,it showed that the formation of osteclasts was restrained intensively by HPDLfs at primary stage with the action of rhBMP2.With the action time prolonging,the restraining effect was changed into the activation effect.