中国组织工程研究与临床康复
中國組織工程研究與臨床康複
중국조직공정연구여림상강복
JOURNAL OF CLINICAL REHABILITATIVE TISSUE ENGINEERING RESEARCH
2009年
31期
6189-6192
,共4页
背景:角膜移植后40%～60%的患者发生了角膜新生血管,移植后的角膜新生血管加速了角膜免疫排斥反应的发生.抑制角膜移植后的角膜新生血管有利于延长植片的存活时间,提高角膜移植手术的成功率.目的:探讨强力霉素对角膜移植后角膜新生血管的抑制作用.设计、时间及单位:随机对照,动物实验观察,于2007 03/08在中山大学中山眼科中心(眼科学国家重点实验室,2006DA1050541)完成.材料:健康清洁级SD大鼠48只作为角膜移植受体,Wistar大鼠24只作为供体,受体眼为右眼,供体眼为双眼.CD31-PE荧光抗体由美国Sigma公司提供,VEGF ELISA试剂盒由美国RapidBio公司提供.方法:建立大鼠同种异体角膜移植模型,48只受体鼠随机分为2组:盐水对照组和强力霉素用约组,24只/组.术前20min以1%阿托品滴眼液散瞳,植片直径为2.75 mm,植床直径为2.5 mm.术后盐水对照组右眼结膜囊内滴用生理眼水,3次/d;强力霉素用药组右眼结膜囊内滴用1%强力霉素眼药水,3次/d:直至术后30d.主要观察指标:全角膜免疫荧光法检测角膜移植后角膜新生血管的发展,ELISA动态检测角膜组织中VEGF蛋白的表达.结果:①强力霉素用药组植片存活(20.67±3.01)d,比盐水对照组(9.67±9.73)d明显延长(P＜0.01).②角膜移植后3,7,14d盐水对照组新生血管面积相对百分比分别为(4.00±1.00)%,(14.33±4.04)%,(31.33±3.51)%;强力霉素用药组新生血管面积相对百分比分别为(1.67±1.15)%,(4.67±1.53)%,(18.33±1.53)%.其中7,14d时两组比较差异显著(P＜0.05).③角膜移植后强力霉素用药组血管内皮生长因子蛋白水平均低于盐水对照组(P＜0.01),.角膜移植后3,7,14d,盐水对照组血管内皮生长因子蛋白水平为(541.00±75.44),(960.00±90.14),(976.00±130.41)ng/g:强力霉索用药组为(115.33±9.29),(239.00±41.62),(361.00±65.20)ng/g,各时间段比较差异显著(P＜0.05).结论:强力霉素能抑制角膜移植后角膜新生血管的生长,延长植片的生存时间.
揹景:角膜移植後40%～60%的患者髮生瞭角膜新生血管,移植後的角膜新生血管加速瞭角膜免疫排斥反應的髮生.抑製角膜移植後的角膜新生血管有利于延長植片的存活時間,提高角膜移植手術的成功率.目的:探討彊力黴素對角膜移植後角膜新生血管的抑製作用.設計、時間及單位:隨機對照,動物實驗觀察,于2007 03/08在中山大學中山眼科中心(眼科學國傢重點實驗室,2006DA1050541)完成.材料:健康清潔級SD大鼠48隻作為角膜移植受體,Wistar大鼠24隻作為供體,受體眼為右眼,供體眼為雙眼.CD31-PE熒光抗體由美國Sigma公司提供,VEGF ELISA試劑盒由美國RapidBio公司提供.方法:建立大鼠同種異體角膜移植模型,48隻受體鼠隨機分為2組:鹽水對照組和彊力黴素用約組,24隻/組.術前20min以1%阿託品滴眼液散瞳,植片直徑為2.75 mm,植床直徑為2.5 mm.術後鹽水對照組右眼結膜囊內滴用生理眼水,3次/d;彊力黴素用藥組右眼結膜囊內滴用1%彊力黴素眼藥水,3次/d:直至術後30d.主要觀察指標:全角膜免疫熒光法檢測角膜移植後角膜新生血管的髮展,ELISA動態檢測角膜組織中VEGF蛋白的錶達.結果:①彊力黴素用藥組植片存活(20.67±3.01)d,比鹽水對照組(9.67±9.73)d明顯延長(P＜0.01).②角膜移植後3,7,14d鹽水對照組新生血管麵積相對百分比分彆為(4.00±1.00)%,(14.33±4.04)%,(31.33±3.51)%;彊力黴素用藥組新生血管麵積相對百分比分彆為(1.67±1.15)%,(4.67±1.53)%,(18.33±1.53)%.其中7,14d時兩組比較差異顯著(P＜0.05).③角膜移植後彊力黴素用藥組血管內皮生長因子蛋白水平均低于鹽水對照組(P＜0.01),.角膜移植後3,7,14d,鹽水對照組血管內皮生長因子蛋白水平為(541.00±75.44),(960.00±90.14),(976.00±130.41)ng/g:彊力黴索用藥組為(115.33±9.29),(239.00±41.62),(361.00±65.20)ng/g,各時間段比較差異顯著(P＜0.05).結論:彊力黴素能抑製角膜移植後角膜新生血管的生長,延長植片的生存時間.
배경:각막이식후40%～60%적환자발생료각막신생혈관,이식후적각막신생혈관가속료각막면역배척반응적발생.억제각막이식후적각막신생혈관유리우연장식편적존활시간,제고각막이식수술적성공솔.목적:탐토강력매소대각막이식후각막신생혈관적억제작용.설계、시간급단위:수궤대조,동물실험관찰,우2007 03/08재중산대학중산안과중심(안과학국가중점실험실,2006DA1050541)완성.재료:건강청길급SD대서48지작위각막이식수체,Wistar대서24지작위공체,수체안위우안,공체안위쌍안.CD31-PE형광항체유미국Sigma공사제공,VEGF ELISA시제합유미국RapidBio공사제공.방법:건립대서동충이체각막이식모형,48지수체서수궤분위2조:염수대조조화강력매소용약조,24지/조.술전20min이1%아탁품적안액산동,식편직경위2.75 mm,식상직경위2.5 mm.술후염수대조조우안결막낭내적용생리안수,3차/d;강력매소용약조우안결막낭내적용1%강력매소안약수,3차/d:직지술후30d.주요관찰지표:전각막면역형광법검측각막이식후각막신생혈관적발전,ELISA동태검측각막조직중VEGF단백적표체.결과:①강력매소용약조식편존활(20.67±3.01)d,비염수대조조(9.67±9.73)d명현연장(P＜0.01).②각막이식후3,7,14d염수대조조신생혈관면적상대백분비분별위(4.00±1.00)%,(14.33±4.04)%,(31.33±3.51)%;강력매소용약조신생혈관면적상대백분비분별위(1.67±1.15)%,(4.67±1.53)%,(18.33±1.53)%.기중7,14d시량조비교차이현저(P＜0.05).③각막이식후강력매소용약조혈관내피생장인자단백수평균저우염수대조조(P＜0.01),.각막이식후3,7,14d,염수대조조혈관내피생장인자단백수평위(541.00±75.44),(960.00±90.14),(976.00±130.41)ng/g:강력매색용약조위(115.33±9.29),(239.00±41.62),(361.00±65.20)ng/g,각시간단비교차이현저(P＜0.05).결론:강력매소능억제각막이식후각막신생혈관적생장,연장식편적생존시간.
BACKROUND:Corneal hemangiogenesis occurs in 40%-60%patients after keratoplasty.Blood vessel is one of the high risk factors for corneal immunological rejection.To inhibit corneal hemangiogenesis would prolong the survival time of the grafts and promote the successful rate of the keratoplasty.OBJECTIVE:To explore the inhibitive effects of doxycycline on corneal angiogenesis after keratoplasty.DESIGN,TIME AND SETTING:A randomized controlled animal experiment was performed at the State Key Laboratory of Ophthalmology(No.2006DA105054),Zhongshan Ophthalmic Center,Sun Yat-sen University from March to August 2007.MATERIALS:A total of 48 healthy dean Sprague Dawley rats served as recipients(right eye)and 24 Wistar rats as donors(both eyes).CD31-PEfluorescent antibody was obtained from Sigma,USA.Sandwich enzyme-linked immunosorbent assay(ELISA)kit for vascular endothelial growth factor(VEGF)was brought from RapidBio,USA.METHODS:Corneal allogenic transplantation models were established in rats.Recipients were equally and randomly divided into 2 groups:saline control group and doxycycline group.Twenty minutes prior to surgery,mydriasis was performed using 1%atropine,with a diameter of 2.75 mm of implant and 2.5 mm of implant bed.In the saline control group,conjunctiva of the right eye received saline,three times a day,following surgery.In the doxycycline group,conjunctiva of the right eye received 1%doxycycline,three times a day,till 30 days following surgery.MAIN OUTCOME MEASURES:The following parameters were measured:corneal angiogenesis using immunofluorescence,expression of VEGF protein by using ELISA.RESULTS:Compared with the survival time of saline control group[(9.67±2.73)days],the mean survival time of doxycycline group[(20.67±3.01)days]was significantly prolonged(P＜0.01).The mean percentages of neovascularized corneal area in the saline control group were(4.00±1.00)%,(14.33±4.04)%,(31.33±3.51)%at 3,7 and 14 days after keratoplasty,respectively.The mean percentages of neovascularized corneal area in the doxycycline group were(1.67±1.15)%,(4.67±1.53)%,(18.33±1.53)%at the same time point respectively.Compared with the saline control group,the mean percentages of neovascularized corneal area of the doxycycline group was significantly reduced at 7 and 14 days after keratoplasty(P ＜0.05).The expression of VEGF in the saline control group was(541.00±75.44)pg/mg,(960.00±90.14)pg/mg,(976.00±130.41)pg/mg at 3,7 and 14 days after keratoplasty,respectively,while expression of VEGF in the doxycycline group was(115.33±9.29)pg/mg,(239.00±41.62)pg/mg,(361.00±65.20)pg/mg,respectively.The difference of VEGF expression at all time points between the two groups was significant (P＜0.05).CONCLUSION:Doxycycline has a significant effect in inhibiting corneal angiogenesis and prolonging survival time of implants after keratoplasty.
