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

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 gingivalfibroblasts,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 matallo-proteinase-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50μg/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、100μg/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 quantitativepolymerase chain reaction,QPCR)检测细胞MMP-1、MMP-8的蛋白和mRNA表达。结果 100μg/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。

Effects of advanced glycation end products on the expression of matrix matelloproteinases in human gingival fibroblasts

Objective To explore the role of （advanced glyeation end products（AGEs）in modulating the expression of matrix matilloproteinase-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.HSA50 μg/mL stimulation group（negative control group）and AGE-modified human serum albumin（AGE-HSA）0.5,5.50and 100μg/mL stimulation groups.After 24,48and 72 hours of culturing,MMP-1and MMP-8 protein and mRNA expression were detected by enzyme-lindked immunosorbent assay （ELISA）and real-time quantitative polymerase chain reaction（QPCR）method respectively.Results At 24,48and 72 hours,the level of MMP-1 in 100μg/mL AGE-HSA group was significantly gradually higher than those in HSA group,blank control group and 0.5μg/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 SGEs may promote synthesis of MMP-1.SGEs may play an important role in periodontal tissue destruction by promoting extracellular collagen degradation.AGEs had no significant effeet on the expression of MMP-8 in eultured HGF according to our results.