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目的:研究脂多糖(lipopolysaccharide,LPS)对大鼠牙髓细胞牙本质涎磷蛋白(dentin sialophosphoprotein,DSPP)、骨涎蛋白(bone sialoprotein,BSP)及碱性磷酸酶(alkaline phosphatase,ALP)表达的影响。方法:采用组织块法获得大鼠牙髓细胞,体外常规培养并进行鉴定,以含0.1、1、10、100和10000 ng/mL牙龈卟啉单胞菌(Porphyromonas gingivalis,P.g)的LPS作用牙髓细胞1、3、5 d,用实时定量PCR检测DSPP、ALP、BSP mRNA表达的变化,采用SPSS17.0软件包对数据进行统计学分析。结果:镜下贴壁后的细胞形态多样,多呈成纤维样细胞形态,还有部分多角形细胞,胞质突起。实时定量PCR结果显示,与对照组相比,1、10 ng/mL LPS组大鼠牙髓细胞DSPP、ALP、BSP的mRNA表达增高,100、10000 ng/mL LPS组DSPP、ALP、BSP的mRNA表达均降低;在1、3、5 d时,1、10、100和10000 ng/mL LPS组mRNA表达逐渐减少。0.1 ng/mL LPS对mRNA的表达无显著影响。3种因子呈现相似的表达变化趋势。结论:低剂量P.g LPS能促进牙髓细胞ALP、BSP、DSPP的表达,高剂量时则抑制ALP、BSP、DSPP的表达;随着培养时间的延长,促进作用逐渐减弱,抑制作用逐渐增强。
Objective: To investigate the effects of lipopolysaccharide (LPS) on the expression of dentin sialophosphoprotein (DSPP), bone sialoprotein (BSP) and alkaline phosphatase (ALP) in rat dental pulp cells influences. METHODS: Rat dental pulp cells were obtained by tissue block method and cultured in vitro. The dental pulp cells were stained with 0.1, 1, 10, 100 and 10000 ng / mL Porphyromonas gingivalis (Pg) The mRNA of DSPP, ALP and BSP was detected by real-time quantitative PCR at 1, 3 and 5 days after transplantation. The data were analyzed by SPSS17.0 software package. Results: The morphology of the cells adhered to each other under microscope was multifarious. Most of them were fibroblast-like cells with some polygonal cells and cytoplasm protruding. Real-time quantitative PCR results showed that compared with the control group, mRNA expression of DSPP, ALP and BSP in 1, 10 ng / mL LPS group increased, DSPP, ALP and BSP mRNA in 100 and 10000 ng / mL LPS group 1, 1, 100, 100 and 10000 ng / mL LPS group. 0.1 ng / mL LPS had no significant effect on mRNA expression. Three kinds of factors showed a similar trend of expression. CONCLUSION: Low dose P.gl LPS can promote the expression of ALP, BSP and DSPP in dental pulp cells, and inhibit the expression of ALP, BSP and DSPP at high dose. With the prolongation of culture time, the promotion effect is weakened and the inhibitory effect is gradually enhanced.