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目的:研究脂多糖(lipopolysaccharide,LPS)对成牙本质细胞(odontoblast-lineage cells,OLCs)细胞连接的影响。方法:在细胞培养体系中加入不同浓度LPS(10、100、1 000 ng/mL)作用于OLCs,并以条件培养基(10 ng/mL LPS)中同时加入NF-κB抑制剂吡咯烷二硫代氨基甲酸酯(Pyrrolidinedithiocarbamate,PDTC)和LPS组为对照;分别培养0.5、1、2、6 h各时间点采用Western blot观察细胞连接蛋白E-cadherin水平的变化;以实时定量PCR观察细胞连接蛋白ZO-1、occludin、claudin-1和E-cadherin mRNA水平的改变。结果:经过LPS处理后,ZO-1、occludin、claudin-1和E-cadherin的mRNA表达水平明显降低,且在一定的作用时间范围内呈时间依赖性(P<0.05),但LPS在浓度10~1 000 ng/mL之间作用无明显差异(P>0.05)。同时,E-cadherin在蛋白水平随着LPS作用时间的延长其表达水平逐渐降低,而当加入NF-κB抑制剂PDTC时则能抑制这种效应。结论:LPS能够下调成牙本质细胞细胞连接蛋白ZO-1、occludin、claudin-1、E-cadherin mRNA的表达水平,以及E-cadherin的蛋白表达水平,对NF-κB的抑制能在一定水平上影响上述过程,表明NF-κB参与了LPS对成牙本质细胞连接的破坏。
Objective: To study the effect of lipopolysaccharide (LPS) on the cell junctions of odontoblast-lineage cells (OLCs). Methods: Different concentration of LPS (10,100,1 000 ng / mL) was added to OLCs in cell culture system, and NF-κB inhibitor pyrrolidine disulfide The cells were treated with PDTC and LPS for 0.5, 1, 2 and 6 h respectively. Western-blotting was used to observe the changes of E-cadherin level. Real-time quantitative PCR Changes of protein ZO-1, occludin, claudin-1 and E-cadherin mRNA levels. Results: After treated with LPS, mRNA expression of ZO-1, occludin, claudin-1 and E-cadherin decreased significantly and was time-dependent in a time-dependent manner (P <0.05) ~ 1 000 ng / mL no significant difference between the role (P> 0.05). At the same time, the expression level of E-cadherin gradually decreased at the protein level with the prolongation of LPS treatment, while the effect was inhibited when adding NF-κB inhibitor PDTC. CONCLUSION: LPS can down-regulate the mRNA expression of connexin ZO-1, occludin, claudin-1 and E-cadherin, and the expression of E-cadherin in dentate cells, and inhibit the expression of E-cadherin to a certain extent Affect the above process, indicating that NF-κB is involved in the destruction of LPL on odontoblasts.