CAJ | 학술논문

目的探讨改良钛支架人工角膜植入兔眼术后,角膜组织基质金属蛋白酶( MMP-2)及其抑制物(TIMP-2)的表达与角膜溶解的关系.方法实验研究.人工角膜支架改良:钛支架表面喷砂后用羟基磷灰石涂层(HA/SB-Ti).将20只正常新西兰白兔分3组每组6只,其中A组、B组右眼角膜板层分别植入改良支架和对照支架(SB-Ti),C组仅做角膜板层切口,余2只为D组做空白对照.同样方法分组20只兔碱烧伤角膜模型,分别为E、F、G、H组.1个月、3个月后取材行免疫组织化学检测MMP-2、TIMP-2在不同组别的表达；为去除碱烧伤因素影响,另取24只正常动物模型改良支架植入后按时间分组,分别为2周组、1月组、3月组、5月组,每组6只,2只正常动物为空白对照组,用实时定量聚合酶链反应(RT-PCR)、免疫印迹法(Western blot)检测MMP-2、TIMP-2mRNA及蛋白的表达.5月组动物及2只碱烧伤模型动物,支架植入3个月后植入镜柱.实验数据采用t检验进行统计学分析.结果 F组有一例出现角膜溶解的并发症.免疫组织化学检查结果:A与B及E与F比较MMP-2表达有统计学意义(t=12.05,2.93,12.00,3.78；P＜0.05).于2周、1个月、3个月、5个月取材后,做RT-PCR、Western blot检测,MMP-2 mRNA及MMP-2蛋白表达从支架植入开始升高,1个月达到高峰,然后下降,仅2周与3个月MMP-2蛋白表达比较无统计学意义(t=2.104,P＞0.05).而TIMP-2 mRNA及TIMP-2蛋白表达在支架植入后有波动,然后逐渐升高,TIMP-2 mRNA除1个月与正常角膜组比较无统计学意义(t=1.878,P=0.0972).结论改良后的人工角膜由于生物活性提高抑制了MMP-2的过度表达,MMP-2、TIMP-2表达随时间的变化规律可能为临床治疗并发症提供新的方法.
목적 탐토개량태지가인공각막식입토안술후,각막조직기질금속단백매( MMP-2)급기억제물(TIMP-2)적표체여각막용해적관계.방법 실험연구.인공각막지가개량:태지가표면분사후용간기린회석도층(HA/SB-Ti).장20지정상신서란백토분3조매조6지,기중A조、B조우안각막판층분별식입개량지가화대조지가(SB-Ti),C조부주각막판층절구,여2지위D조주공백대조.동양방법분조20지토감소상각막모형,분별위E、F、G、H조.1개월、3개월후취재행면역조직화학검측MMP-2、TIMP-2재불동조별적표체；위거제감소상인소영향,령취24지정상동물모형개량지가식입후안시간분조,분별위2주조、1월조、3월조、5월조,매조6지,2지정상동물위공백대조조,용실시정량취합매련반응(RT-PCR)、면역인적법(Western blot)검측MMP-2、TIMP-2mRNA급단백적표체.5월조동물급2지감소상모형동물,지가식입3개월후식입경주.실험수거채용t검험진행통계학분석.결과 F조유일례출현각막용해적병발증.면역조직화학검사결과:A여B급E여F비교MMP-2표체유통계학의의(t=12.05,2.93,12.00,3.78；P＜0.05).우2주、1개월、3개월、5개월취재후,주RT-PCR、Western blot검측,MMP-2 mRNA급MMP-2단백표체종지가식입개시승고,1개월체도고봉,연후하강,부2주여3개월MMP-2단백표체비교무통계학의의(t=2.104,P＞0.05).이TIMP-2 mRNA급TIMP-2단백표체재지가식입후유파동,연후축점승고,TIMP-2 mRNA제1개월여정상각막조비교무통계학의의(t=1.878,P=0.0972).결론 개량후적인공각막유우생물활성제고억제료MMP-2적과도표체,MMP-2、TIMP-2표체수시간적변화규률가능위림상치료병발증제공신적방법.
Objectives To investigate the expression of matrix metalloproteinase-2 (MMP-2) and Tissue inhibior of matrix metalloproteinase-2 (TIMP-2) in rabbit corneas implanted with modified titanium skirt of keratoprosthesis in order to explore the potential roles.Methods A total of 20 New Zealand white rabbits with corneal alkali burn in right eye rabbit corneas were divided into three groups.There were 6 animals in each group.Skirt of hydroxyapatite /Sandblast-Titanium and Sandblast-Titanium were inserted into the corneal stroma of rabbits in group A and group B.The group C did not insert skirt as surgical control.2 rabbits were as normal control D group.A total of 20 New Zealand white rabbits were divided into four groups with the same way.The expression of MMP-2 and TIMP-2 was determined by immunohistochemistry at 1 month,3 months.The expression of MMP-2 and TIMP-2 mRNA level was determined by real time-polymerase chain reaction,and its protein level was determined by western blot.The optical cylinder was implanted to rabbit corneas,which were implanted with modified titanium skirt after 3 months.Results There was one case of corneal dissolution being found in group F.MMP-2 and TIMP-2 immunoreactivities were expressed in the normal comeas,predominantly in the corneal epithelium.After injury,immunoreactivities of both MMP-2 and TIMP-2 were increased notably in the healing corneal epithelium,infiltrating inflammatory cells,stromal fibroblasts and in growing vascular endothelial cells.The expression of MMP-2 was lower in group A and E than that in group B and F after 1 month and 3 months (t =12.05,2.93,12.006,3.781,P ＜ 0.05 ).The Western bolt revealed no significant differences of MMP-2 mRNA between group 3 months and 2 weeks ( t =2.104,P ＞ 0.05 ) ; MMP-2 immunoreactivities were absent or lowly expressed predominantly in the corneal epithelium of normal corneas.The expression of MMP-2,TIMP-2 mRNA level was paralled that of protein level.Conclusions The expression of MMP-2 was lower in the corneal tissue sections of HA/SB-Ti skirt inserted eyes than that in the tissue sections of SB-Ti skirt inserted eyes.The studies of MMP-2,TIMP-2 can provide a new way to prevent the incidence of corneal dissolving after surgery for keratoprosthesis.