糖基化终产物对人牙龈成纤维细胞2种基质金属蛋白酶表达的影响

摘要

目的 研究糖基化终产物(advanced glycation end products,AGEs)对人牙龈成纤维细胞(human gingival fibroblasts,HGF)表达基质金属蛋白酶-1(matrix matelloproteinase-1,MMP-1)和基质金属蛋白酶-8(matrix matalloproteinase-8,MMP-8)的影响,探讨糖尿病加速牙周炎发展的可能机制.方法 将培养的HGF随机分成6组:空白对照组只加入培养液;阴性对照组仅加入含50 μg/mL人血清白蛋白(human serum albumin,HSA)的培养液;4个含AGE-HSA的实验组分别加入含0.5、5、50、100 μg/mL AGE-HSA的培养液.培养24、48、72 h后,分别应用酶联免疫吸附测定法(enzyme-linked immunosorbent assay,ELISA)和实时定量聚合酶链反应(real-time quantitative polymerase chain reaction,QPCR)检测细胞MMP-1、MMP-8的蛋白和mRNA表达.结果 100 μg/mL AGE-HSA组培养24、48和72 h后,MMP-1水平明显高于阴性对照组、空白对照组及0.5μg/mL AGE-HSA组,差异具有统计学意义(P＜0.05).各浓度AGE-HSA组MMP-1 mRNA水平均显著高于阴性对照组,差异有统计学意义(P＜0.05).各浓度AGE-HSA组MMP-8水平与阴性对照组相比差异均无统计学意义(P＞0.05),MMP-8 mRNA表达均为阴性.结论 AGEs可能通过促进HGF合成MMP-1,介导胶原降解,从而加重糖尿病患者的牙周组织破坏,尚不能说AGEs影响MMP-8的表达.%Objective To explore the role of advanced glycation end products (AGEs) in modulating the expression of matrix matelloproteinase-1 (MMP-1) and matrix matelloproteinase-8 (MMP-8) in human gingival fibroblasts (HGF). Methods The cultured HGF cells were randomly divided into six groups: blank control group, HSA 50 μg/mL stimulation group (negative control group) and AGE-modified human serum albumin (AGE-HSA) 0.5, 5, 50 and 100 μg/mL stimulation groups. After 24, 48 and 72 hours of culturing, MMP-1 and MMP-8 protein and mRNA expression were detected by enzyme-linked immunosorbent assay (ELISA) and real-time quantitative polymerase chain reaction ( QPCR) method respectively. Results At 24, 48 and 72 hours, the level of MMP-1 in 100 μg/mL AGE-HSA group was significantly gradually higher than those in HSA group, blank control group and 0. 5 μg/mL AGE-HSA group (P <0.05) . The level of MMP-1 mRNA in each concentration AGE-HSA group was significantly higher than that in HSA group (P < 0. 05). There was no statistically significant difference between the level of MMP-1 in each concentration AGE-HSA group and that in HSA group (P>0. 05). MMP-8 mRNA expression in each concentration AGE-HSA group was negative. Conclusion AGEs may promote synthesis of MMP-1. AGEs may play an important role in periodontal tissue destruction by promoting extracellular collagen degradation. AGEs had no significant effect on the expression of MMP-8 in cultured HGF according to our results.