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目的研究不同浓度的牙龈卟啉单胞菌(Porphyromonas gingivalis,P.gingivalis)脂多糖(Lipopolysaccharide,LPS)调节人牙周膜细胞(hPDLFs)中骨涎蛋白(Bone sialoprotein,BSP)基因表达的影响。方法采用组织块法原代培养hPDLFs,取第4代细胞在含有10%胎牛血清(FCS)的DMEM培养基中培养至完全融合后,分别用0.01、0.1、1、10、100mg/L P.g.LPS作用hPDLFs 10h,实时荧光定量PCR检测骨涎蛋白基因(BSP mRNA)的表达水平。结果组织块法hPDLFs原代培养5~8d后,可见典型细长梭形细胞从组织块边缘呈放射状游离出来,第4代传代细胞伸展贴壁良好;0.01、0.01mg/L P.g.LPS刺激hPDLFs10h后BSP mRNA表达略有升高,但差异无统计学意义(P>0.05);而10、100 mg/L P.g.LPS刺激BSP mRNA表达显著降低,差异有统计学意义(P<0.01)。结论不同浓度的P.g.LPS对人hPDLFs中BSP基因表达有不同影响,高浓度P.g.LPS明显降低BSP基因的表达水平。
Objective To study the effects of different concentrations of Porphyromonas gingivalis (Lipuracein) on lipopolysaccharide (LPS) in regulating the expression of Bone Sphoprotein (BSP) gene in human periodontal ligament cells (hPDLFs). Methods Primary cultured hPDLFs were cultured by tissue block method. The fourth passage cells were cultured in DMEM medium containing 10% fetal bovine serum (FCS) until they were completely fused and then treated with 0.01, 0.1, 1, 10 and 100 mg / L Pg The effect of LPS on hPDLFs was detected by real-time fluorescence quantitative PCR for detecting the expression of bone sialoprotein (BSP) mRNA. Results After primary culture of hPDLFs by tissue block method for 5 ~ 8 days, typical slender spindle cells released radially from the edge of the tissue block, and the passage cells of the 4th passage adhered well. 0.01, 0.01 mg / L PgLPS stimulated hPDLFs for 10 h BSP mRNA expression was slightly increased, but the difference was not statistically significant (P> 0.05); while BSP mRNA expression was significantly reduced by PgLPS 10,100 mg / L (P <0.01). Conclusions Different concentrations of P.g.LPS have different effects on BSP gene expression in human hPDLFs. High concentration of P.g.LPS significantly reduces BSP gene expression.