糖基化终末产物对人牙龈成纤维细胞炎症因子分泌及P65表达的影响

Effects of advanced glycation end products on secretion of inflammatory factors and expression of P65 in human gingival gibroblasts

目的:研究糖基化终末产物(AGEs)对人牙龈成纤维细胞炎症因子分泌的影响及其可能的机制。方法:用体外酶消化结合组织块法培养人牙龈成纤维细胞;将细胞分为对照组和实验组,对照组为正常培养液培养的人牙龈成纤维细胞;实验组为含AGEs终浓度分别为1、5、10、20μL/mL培养液培养的人牙龈成纤维细胞;Real time PCR检测炎症因子IL-6、肿瘤坏死因子TNF-α、NF-κB信号通路关键分子P65mRNA的表达情况。结果:酶消化加组织块法培养出来的牙龈成纤维细胞大多呈长梭形,胞液丰满;Real time PCR检测结果表明,与对照组相比,不同浓度AGEs刺激下的牙龈成纤维细胞IL-6表达量均升高,5、10、20μL/mL浓度AGEs刺激下TNF-α、P65的mRNA表达量升高,差异有统计学意义(P<0.05)。结论:糖基化终末产物可以导致牙龈成纤维细胞炎症因子的表达升高,其机制可能是通过激活NF-κB信号通路,从而影响了糖尿病患者的牙龈健康。

Objective:To study the effect of advanced glycation end products(AGEs)on the secretion of inflammatory factors in human gingival fibroblasts and its possible mechanism.Methods:Human gingival fibroblasts were cultured in vitro using enzyme digestion combined with tissue block technique,and divided into control group and experimental group.Human gingival fibroblasts in the control group were cultured in conventional medium,and those in the experimental group were cultivate in 1,5,10 and 20μL/mL media with AGEs end concentration,respectively.Real time PCR was performed to detect the inflammatory factor IL-6,TNF-αand p65 mRNA expression.Results:Most of the gingival fibroblasts cultured by enzyme digestion and tissue block technique presented with long fusiform and full cytoplasm.The results of Real time PCR showed that mRNA expression of IL-6 as well as TNF-αand p65 mRNA expression in gingival fibroblasts stimulated by diverse dose of AGEs or by 5,10 and 20μL/mL AGEs was notably increased as compared to the control group.The difference was significant(P<0.05).Conclusion:Advanced glycation end products can up-regulate the expression of inflammatory factors in gingival fibroblasts,which may be related to the activation of NF-κB signaling pathway,thus affecting the gingival health of diabetic patients.