【作者】 雍翔智； 陈怡； 曾启新； 陶人川；

【Author】 Yong Xiangzhi;Chen Yi;Zeng Qixin;Tao Renchuan;Department of Periodontics and Oral Medicine, College of Stomatology, Guangxi Medical University;

【机构】 广西医科大学口腔医学院牙周粘膜科；

【摘要】 目的:通过TNF-α+IFN-γ刺激人口腔黏膜上皮细胞(hOMEC)建立炎症模型,观察炎症模型中地塞米松(Dexamethasone,Dex)对于口腔黏膜上皮防御素(hunman Beta-defensins,hBDs)及Toll(Toll-like receptors,TLR)样受体表达的影响。方法:选用不同浓度的Dex(0.1,1,10μM)预处理第二代hOMEC12h后,再用50ng/ml (TNF-α+IFN-γ)刺激细胞24h,RT-PCR检测处理前后细胞TLR2、hBD-2、hBD-3的mRNA相对表达量,取贴壁细胞,提取细胞总蛋白行Western Blot(WB)检测TLR2蛋白的表达情况。ELISA检测细胞上清液中hBD-2蛋白水平。结果:不同浓度DEX均能使TNF-α+IFN-γ诱导的hOMEC hBD-2的mRNA相对表达量下降,且随Dex浓度增加而降低,在浓度为10μM时,差异有统计学意义(P<0.05)。在hBD-2蛋白水平,不同浓度DEX均能使hBD-2表达量下降(P<0.05),且不同浓度的Dex作用下,hBD-2表达量之间差异无明显统计学意义(P>0.05)而在任何浓度的Dex作用下,hBD-3的mRNA相对表达量与模型组相比,差异无统计学意义(P>0.05)。TNF-α+IFN-γ能使TLR2的mRNA相对表达量明显升高,在Dex浓度为0.1,1μM时TLR2的mRNA水平有轻微升高,但差异无统计学意义(P>0.05)。0.1,1,10μM的Dex均能使TLR2的蛋白表达量下降,且低浓度的DEX下调TLR2的蛋白表达量最明显。结论:DEX能抑制hOMEC炎症模型中hBD-2的表达,其机制可能与下调TLR2信号通路表达有关。更多还原

【Abstract】 Objective: To detect the influence of dexamethasone(Dex) on the expression of toll like receptors and hBD-2 and hBD-3 hOMEC induced by ininflammatory factor in human oral mucosal epithelial cells(hOMEC). Methods: After pretreated with different concentrations of Dex(0.1,1,10μM)for 12 h, the second generation of hOMEC was induced by 50ng/ml(TNF-alpha + IFN-gamma) for 24 h. RT-PCR was used to detect the relative expression of TLR2, hBD-2 and hBD-3 mRNA in cells. The total protein was extracted from anchorage-dependent cells and detect the expression of TLR2 protein by Western Blot(WB). ELISA was used to detect the level of hBD-2 protein in the cell supernatant. Results: Different concentrations of Dex could decrease the relative expression of hBD-2 mRNA induced by inflammatory factors in hOMEC.And as the concentration of Dex increased, the more obvious hBD-2 mRNA decreased. The difference was statistically significant when the concentration was 10μM(P<0.05). At the level of hBD-2 protein, all concentrations of Dex could decrease the expression of hBD-2(P<0.05), and there was no significant difference between different concentrations of Dex in the expression of hBD-2(P>0.05).However, At any concentration of Dex, the relative expression of hBD-3 mRNA was not statisticallysignificant compared with that of the model group(P>0.05).TNF-α+IFN-γ could increase the relative expression of mRNA of TLR2, and the mRNA level of TLR2 increased slightly at concentration of 0.1,1 μM Dex, but the difference was not statistically significant(P>0.05). The Dex of 0.1,1,10μM could decrease the protein expression of TLR2, and the low concentration of Dex down regulated TLR2 protein expression was the most obvious. Conclusion: DEX could inhibit the expression of the hBD-2 by inflammatory factors in hOMEC by suppressing the TLR2 signaling pathway.更多还原