IL1β干预下TRA6在牙周膜成纤维细胞中表达的研究.docx

坐苎堡型奎兰竺主兰竺堡兰Abst 坐苎堡型奎兰竺主兰竺堡兰 Abst ract IObjectivel：To isolate and culmre the human periodontal ligament fibroblasts(HPLFs)， construct the cell model in vitro；and research the expression of tumor necrosis factor receptor -associated factor一6(TRAF6)in HPLFs by the interference of IL．1 13，and provide new document to the research ofthe effect ofcytokine network to cells． IMethod]：The primary cells were isolated from human periodontal ligament and cultured bv tissue explant culture technique，then passaged when the cells grew in the bottom of culture flasks completely．Morphological analysis and immunohiStochemistry analysis were used to characterize the cell lineage．Choose the fifth to eighth passage and stable growth of cells．And Use IL-I 13 of Ong／ml、0．I ng／ml、O．5ng／ml、l ng／ml、5ng／ml、1 0ng／ml on cells intervention respectively· Using rt,朗verse transcription·-polymerase chain reaction(RT-PCR) and immunohistochemical methods to detect the expression of TRAF6，and collect the test results，include image and data by computer image analysis system．The experiment data using single—factor variance Analysis for statistical analysis，to observe the expression and Iocation of TRAF6 in different concentrations of IL一1 13 intervention． IResult]：(1)The HPLFs were cultured and passaged successfully．After the fifth passage，cells 伊ew prosperity and show long spindle appearance,are a typical fibroblast．1ike morphology． Through immunocytochemical analysis，the cells had a positive reaction to antibodies against vimentin， and a negatwe reaction to antibodies against ceratin． It certified the cells were fibroblasts from mesoblast，and in line with human periodontal ligament fibroblasts in vitro biological properties．(2)The results of RT-PCR and immunohistochemistrv showed that：in the gene and protein level，TRAF6，in the normal HPLFs shows a negative reaction,but shows a posmve reaction in the cytoplasm of the cell by IL一1 p intervention,and the intensity of its expression of IL一1 13 inter