中华口腔医学杂志
中華口腔醫學雜誌
중화구강의학잡지
Chinese Journal of Stomatology
2012年
10期
599-604
,共6页
宋爱梅%侯超%陈家芳%孙静%田恬%李纾
宋愛梅%侯超%陳傢芳%孫靜%田恬%李紓
송애매%후초%진가방%손정%전념%리서
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
牙週膜%成纖維細胞%缺氧%基質金屬蛋白酶%組織金屬蛋白酶抑製劑
아주막%성섬유세포%결양%기질금속단백매%조직금속단백매억제제
Periodontal ligament% Fibroblasts% Anoxia% Matrix metalloproteinases% Tissue inhibitors of matrix metalloproteinases
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48 h(分别为12、24、48 h缺氧组)；常氧组细胞在20％O2、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、48 h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-1,2 mRNA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-1,2 mRNA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、48 h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.
目的 探討缺氧對人牙週膜成纖維細胞基質金屬蛋白酶(matrix metalloproteinase,MMP)和組織金屬蛋白酶抑製物(tissue inhibitors of matrix metalloproteinase,TIMP) mRNA錶達的影響,以期為牙週組織脩複再生機製研究提供依據.方法 組織塊法培養人牙週膜成纖維細胞,分為缺氧組和常氧組(對照組).缺氧組細胞分彆于1％O2、5％ CO2、94％N2的37℃培養箱中培養12、24和48 h(分彆為12、24、48 h缺氧組)；常氧組細胞在20％O2、5％ CO2、75％N2的37℃培養箱中培養.採用反轉錄聚閤酶鏈反應技術檢測MMP和TIMP相關基因的錶達.缺氧組與常氧組間均數比較採用t檢驗,多組間比較採用單因素方差分析,兩兩比較採用LSD檢驗.結果 12、24、48 h缺氧組MMP-2 mRNA的錶達呈明顯遞增趨勢(分彆為0.769±0.038、0.793±0.043、0.865±0.047),均顯著高于常氧組(0.294±0.117),P＜0.01；12h缺氧組TIMP-1,2 mRNA的錶達(分彆為1.870±0.645、0.862±0.043)均有一過性升高,均分彆顯著高于常氧組TIMP-1,2 mRNA的錶達(分彆為0.816±0.043、0.426±0.097),P＜0.05.12h缺氧組MMP-1/TIMP-1 mRNA的比值(0.392±0.206)顯著低于常氧組(0.859 ±0.126),P＜0.05；24、48 h缺氧組MMP-2/TIMP-2 mRNA的比值(分彆為1.091±0.207、1.264±0.377)均顯著高于常氧組(0.695±0.255),P＜0.05.結論 缺氧可以導緻MMP-1、MMP-2、TIMP-1、TIMP-2 mRNA的錶達,併使MMP-2 mRNA與TIMP-2 mRNA的比值失衡,這可能是缺氧導緻牙週炎患者牙週組織破壞加重的機製之一.
목적 탐토결양대인아주막성섬유세포기질금속단백매(matrix metalloproteinase,MMP)화조직금속단백매억제물(tissue inhibitors of matrix metalloproteinase,TIMP) mRNA표체적영향,이기위아주조직수복재생궤제연구제공의거.방법 조직괴법배양인아주막성섬유세포,분위결양조화상양조(대조조).결양조세포분별우1％O2、5％ CO2、94％N2적37℃배양상중배양12、24화48 h(분별위12、24、48 h결양조)；상양조세포재20％O2、5％ CO2、75％N2적37℃배양상중배양.채용반전록취합매련반응기술검측MMP화TIMP상관기인적표체.결양조여상양조간균수비교채용t검험,다조간비교채용단인소방차분석,량량비교채용LSD검험.결과 12、24、48 h결양조MMP-2 mRNA적표체정명현체증추세(분별위0.769±0.038、0.793±0.043、0.865±0.047),균현저고우상양조(0.294±0.117),P＜0.01；12h결양조TIMP-1,2 mRNA적표체(분별위1.870±0.645、0.862±0.043)균유일과성승고,균분별현저고우상양조TIMP-1,2 mRNA적표체(분별위0.816±0.043、0.426±0.097),P＜0.05.12h결양조MMP-1/TIMP-1 mRNA적비치(0.392±0.206)현저저우상양조(0.859 ±0.126),P＜0.05；24、48 h결양조MMP-2/TIMP-2 mRNA적비치(분별위1.091±0.207、1.264±0.377)균현저고우상양조(0.695±0.255),P＜0.05.결론 결양가이도치MMP-1、MMP-2、TIMP-1、TIMP-2 mRNA적표체,병사MMP-2 mRNA여TIMP-2 mRNA적비치실형,저가능시결양도치아주염환자아주조직파배가중적궤제지일.
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 α-minima essential medium(α-MEM) and subcultured at confluence. In the hypoxic groups,cells were incubated in a humidified atmosphere of 1％O2,5％CO2,94％ N2 at 37 ℃ for 12,24 and 48 h,respectively.In the normoxic control group,cells were incubated under normoxic conditions of 20％ O2,5％ CO2,75％ N2.The mRNA expression of MMP and TIMP was measured using reverse transcription-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 hypoxic groups showed a significantly increasing trend.There was significant difference between the hypoxic group and the normoxic 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 control group in the ratio of MMP-1/TIMP-1 mRNA ( P ＜ 0.05 ).The ratio of MMP-2/ TIMP-2 mRNA in the hypoxia group significantly increased compared with normoxic group. There were significant differences between the hypoxic group and the normoxic control group in the ratio of MMP-2/TIMP-2 mRNA(P ＜ 0.05).Conclusions Hypoxia could change the expression of MMP and TIMP mRNA and other relevant growth factors and also lead to the imbalance of MMP-2/TIMP-2 mRNA expression.It is suggested that the imbalance of MMP-2/TIMP-2 expression may be closely correlated with the occurrence and development of periodontal disease and play an important role in the process of periodontal tissue destruction in periodontitis.
