缺氧对人牙周膜成纤维细胞基质金属蛋白酶和组织金属蛋白酶抑制物mRNA表达的影响  

Effect of hypoxia on the expression of matrix metalloproteinase and tissue inhibitors of matrix metalloproteinase mRNA in human periodontal ligament fibroblasts in vitro

作　　者：宋爱梅[1] 侯超[2] 陈家芳[1] 孙静[1] 田恬[1] 李纾[1] 

机构地区：[1]山东大学口腔医学院牙周病科·山东省口腔生物医学重点实验室,济南250012 [2]山东省枣庄市立医院口腔科

出　　处：《中华口腔医学杂志》2012年第10期599-604,共6页Chinese Journal of Stomatology

基　　金：山东省自然科学基金（ZR2010HM050）

摘　　要：目的探讨缺氧对人牙周膜成纤维细胞基质金属蛋白酶（matrix metalloproteinase，MMP）和组织金属蛋白酶抑制物（tissue inhibitors of matrix metalloproteinase，TIMP）mRNA表达的影响，以期为牙周组织修复再生机制研究提供依据。方法组织块法培养人牙周膜成纤维细胞，分为缺氧组和常氧组（对照组）。缺氧组细胞分别于1％O2、5％CO2、94％N2的37℃培养箱中培养12、24和48h（分别为12、24、48h缺氧组）；常氧组细胞在20％02、5％CO2、75％N2的37℃培养箱中培养。采用反转录聚合酶链反应技术检测MMP和TIMP相关基因的表达。缺氧组与常氧组间均数比较采用t检验，多组间比较采用单因素方差分析，两两比较采用LSD检验。结果12、24、48h缺氧组MMP-2 mRNA的表达呈明显递增趋势（分别为0．769±0．038、0．793±0．043、0．865±0．047），均显著高于常氧组（0．294±0．117），P〈0．01；12h缺氧组TIMP-1，2mRNA的表达（分别为1．870±0．645、0．862±0．043）均有一过性升高，均分别显著高于常氧组TIMP-1，2mRNA的表达（分别为0．816±0．043、0．426±0．097），P〈0．05。12h缺氧组MMP-1／TIMP-1 mRNA的比值（0．392±0．206）显著低于常氧组（0．859±0．126），P〈0．05；24、48h缺氧组MMP-2／TIMP-2 mRNA的比值（分别为1．091±0．207、1．264±0．377）均显著高于常氧组（0．695±0．255），P〈0．05。结论缺氧可以导致MMP-1、MMP-2、TIMP-1、TIMP-2 mRNA的表达，并使MMP-2 mRNA与TIMP-2 mRNA的比值失衡，这可能是缺氧导致牙周炎患者牙周组织破坏加重的机制之一。Objective To investigate the effect of hypoxia on the expression of matrix metalloproteinase（MMP） and tissue inhibitors of matrix metalloproteinase （TIMP） in human periodontal ligament fibroblasts（HPDLF）. Methods HPDLF were cultured in a-minima essential medium （ot-MEM） and subcultured at confluence. In the hypoxic groups, cells were incubated in a humidified atmosphere of 1%02,5%CO2,94% N2 at 37 ℃ for 12,24 and 48 h, respectively. In the normoxic control group, ceils were incubated under normoxic conditions of 20% O2, 5% CO2, 75% N2. The mRNA expression of MMP and TIMP was measured using reverse transeription-polymerase chain reaction （RT-PCR）. The data was analyzed by Student＇s t test, one-way ANOVA and LSD test with SPSS 13.0 software package. Results The expression of MMP-2, TIMP-1 and TIMP-2 mRNA in the hypoxia groups was higher than that in control. The expression of MMP-2 mRNA in hypoxie groups showed a significantly increasing trend. There was significant difference between the hypoxie group and the normoxie control group in the expression of MMP-2 mRNA in HPDLF（ P 〈 0.01 ）. The expression of TIMP-1, TIMP-2 mRNA in hypoxic groups of 12 h was momentarily increased. There was significant difference between the hypoxic 12 h group and the normoxic control group in the expression of TIMP-1 ,TIMP-2 mRNA in HPDLF（P 〈 0.05 ）. However, with prolonged hypoxia time, the expression of TIMP-1, TIMP-2 mRNA in hypoxic groups showed a significantly declining trend, there were significant differences between the hypoxic 12, 24 and 48 h group and the normoxic control group in the expression of TIMP-2 mRNA in HPDLF（ P 〈 0.05 ）. The expression of MMP-1 mRNA in hypoxic groups of 12 h was momentarily decreased and then increased after 24 h of hypoxia. There were significant differences between the hypoxic 48 h group and the normoxic control group in the expression of MMP-1 mRNA in HPDLF（P 〈 0.05）. There were significant differences between the hypoxic 12 h group and the normoxic

关 键 词：牙周膜 成纤维细胞 缺氧 基质金属蛋白酶 组织金属蛋白酶抑制剂 

分 类 号：R780.2[医药卫生—口腔医学]