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目的:研究早期生长转录因子(early growth responsive gene-1,Egr-1)对机械应力(mechanical stress,MS)介导人牙周膜细胞(human periodontal membrane cells,hPDLs)炎症因子分泌和表达的影响及可能机制。方法:以实时定量PCR和ELISA法检测MS加载不同时间(6、12、24和48 h)和不同形变率(3%、6%、12%和15%)时炎症因子IL-1β、IL-6、IL-8和IL-11的分泌和表达水平。MS对Egr-1表达的影响采用实时定量PCR和Western印迹法检测,沉默Egr-1对炎症因子的作用采用实时定量PCR和ELISA检测。应用Western印迹法检测Egr-1下调对PTEN/PI3K/Akt信号通路相关蛋白表达的影响。进一步采用20μmol/L PI3K/Akt抑制剂LY294002预处理hPDLs 30 min,研究Egr-1沉默在MS介导炎症因子表达中的可能机制。采用SPSS 11.0软件包对数据进行统计学分析。结果:IL-1β、IL-6、IL-8和IL-11的分泌和mRNA表达随着作用时间和形变率的增加而升高,12%的MS作用24 h后,其分泌程度和表达水平最高。12%的MS加载24 h显著上调Egr-1表达。沉默Egr-1显著抑制MS诱导炎症因子表达。Egr-1沉默下调PTEN表达,上调p-PI3K和p-Akt蛋白表达水平,LY294002预处理可部分阻断Egr-1沉默对炎症因子分泌及表达的抑制作用。结论:沉默Egr-1通过PETN/PI3K/Akt信号通路抑制MS诱导的炎症因子分泌和表达。
AIM: To investigate the effect of early growth responsive gene-1 (Egr-1) on the secretion and expression of inflammatory cytokines in human periodontal membrane cells (hPDLs) mediated by mechanical stress (MS) And possible mechanism. METHODS: Real-time quantitative PCR and ELISA were used to detect the expression of IL-1β, IL-1β at different time points (6, 12, 24 and 48 h) and different deformation rates (3%, 6%, 12% and 15% 6, IL-8 and IL-11 secretion and expression levels. The effect of MS on Egr-1 expression was detected by real-time quantitative PCR and Western blotting. The effect of Egr-1 silencing on inflammatory cytokines was detected by real-time PCR and ELISA. The effect of Egr-1 down-regulation on PTEN / PI3K / Akt signaling pathway-related protein expression was detected by Western blotting. HPDLs were further pretreated with 20μmol / L PI3K / Akt inhibitor LY294002 for 30 min to investigate the possible mechanism of Egr-1 silencing in MS-mediated inflammatory factor expression. Data were analyzed statistically using SPSS 11.0 software package. Results: Secretion and mRNA expression of IL-1β, IL-6, IL-8 and IL-11 increased with time and the rate of degeneration. After 12% MS for 24 h, the secretion and expression of IL- highest. 12% MS loading 24 h significantly upregulated Egr-1 expression. Silencing Egr-1 significantly inhibited MS-induced inflammatory factor expression. Egr-1 silencing down-regulated the expression of PTEN and up-regulated the expression of p-PI3K and p-Akt. LY294002 pretreatment could partially block the inhibitory effect of Egr-1 silencing on the secretion and expression of inflammatory cytokines. Conclusion: Silencing Egr-1 inhibits the secretion and expression of inflammatory cytokines induced by MS through PETN / PI3K / Akt signaling pathway.