目的探讨胞内信号转导分子FHL2蛋白在人牙周膜细胞(hPDLCs)体外矿化过程中的表达。方法体外培养hPDLCs,实验组用矿化诱导液培养,对照组不加诱导液。培养0、14、28d后,茜素红染色检测矿化结节的形成;免疫细胞化学法检测hPDLCs矿化诱导0、14 d时FHL2蛋白的表达;同时采用半定量RT-PCR方法检测hPDLCs矿化诱导0、14、28 d时FHL2 mRNA的表达。结果诱导培养14、28 d后,实验组茜素红染色阳性,矿化结节形成;对照组茜素红染色显示:无明显矿化结节形成。免疫细胞化学结果显示:实验组FHL2蛋白阳性表达。RT-PCR结果显示:与0d比较,hPDLCs矿化培养14和28 d后,FHL2表达均有显著升高(P〈0.01),其中14 d约为0 d的1.4倍。结论 FHL2与hPDLCs的体外矿化过程相关。提示FHL2可能在hPDLCs的分化、矿化过程中发挥作用。
Objective To investigate the expression pattern of FHL2,which is an intracellular signaling transcription molecule during mineralization in cultured human periodontal ligament cells(hPDLCs) in vitro.Methods hPDLCs were cultured in vitro.Test group was cultured with mineral induction media while control group without induction media.0,14,28 days after culture,alizarin red staining was used to measure the mineral nodules formation.Immunocyto-chemistry was used to examine the expression of FHL2 protein 0 day and 14 days after mineral induction.Mean-while,mRNA expression level of FHL2 was analyzed by reverse transcriptase-polymerase chain reaction(RT-PCR) on the 0,14,28 days after induction.Results 14 and 28 days after cultivation,mineral nodules formed and were stained positively with alizarin red staining in test group while no mineral nodule formed in control group.Immunocy-tochemical results indicated that hPDLCs in test group expressed FHL2 positively.According to RT-PCR results,14 and 28 days after mineral induction,the expression levels of FHL2 both increased significantly when compared with 0 day(P0.01),and the expression level at 14 days was 1.4 folds of 0 day.Conclusion FHL2 protein is found to be involved in the in vitro mineralization of hPDLCs.FHL2 protein may play a role in the differentiation and minerali-zation of hPDLCs.