高危角膜移植排斥兔模型的建立及对比研究
고위각막이식배척토모형적건립급대비연구
Comparison among three modeling methods of high-risk rejection corneal transplantation
  • 万方数据
    中华实验眼科杂志 중화실험안과잡지
    CHINESE JOURNAL OF EXPERIMENTAL OPHTHALMOLOGY
    2012年 1期 7-11 ,共5页

    李乃洋%王晓然%万鹏霞%黄明海%武征%梁轩伟%刘颖%葛坚%王智崇

    리내양%왕효연%만붕하%황명해%무정%량헌위%류영%갈견%왕지숭

    器官移植/角膜、高危%动物模型%排斥反应%异种移植 器官移植/角膜、高危%動物模型%排斥反應%異種移植
    기관이식/각막、고위%동물모형%배척반응%이충이식
    Organ transplantation/cornea,high-risk%Animal model%Rejection%Xenotransplantation
    背景 理想的角膜移植排斥动物模型是研究高危角膜移植免疫排斥机制的基础,具有重要的意义. 目的 比较各种建立兔高危角膜移植排斥模型方法的临床特点,探索合适的角膜移植排斥动物模型的建立方法.方法 45只新西兰白兔作为角膜移植受体,并按照造模方法的不同按随机数字表法随机分为缝线组、碱烧伤组和异种移植组,每组15只.分别用在角膜4个象限各间断缝1根5-0丝线法和1 mol/LNaOH碱烧伤法诱导角膜新生血管(CNV),再建立兔同种异体角膜移植；另一组以猫角膜为供体,建立猫-兔异种角膜移植模型.于第2周和第4周观察植片的组织学情况,对3个组角膜植片裂隙灯下观察植片排斥反应、炎症和新生血管,对植片水肿程度及炎症指数(IF)进行评分,根据角膜混浊、水肿及新生血管合计值计算排斥指数(RI).用免疫组织化学法检测CD4+T细胞和CD8+T细胞在植片组织中的表达. 结果 缝线组、碱烧伤组和异种移植组分别有14、15、15只兔完成穿透角膜移植术.术后2周,3个组IF中位数分别为0.556、0.778、0.222,差异有统计学意义(H=25.736,P=0.000),异种移植组IF值低于缝线组和碱烧伤组,差异均有统计学意义(Z=3.841、3.993,P=0.000),缝线组IF值低于碱烧伤组,差异有统计学意义(Z=3.568,P=0.000).术后2周,3个组RI中位数分别为2、6、3,差异有统计学意义(H=22.432,P=0.000),异种移植组RI高于缝线组而低于碱烧伤组,差异均有统计学意义(Z=2.373,P=0.018；Z=3.936,P=0.000),缝线组RI低于碱烧伤组,差异有统计学意义(Z=3.729,P=0.000).3个组植片存活时间分别为(17.9±2.0)、(13.4±2.4)、(15.5±2.0)d,差异有统计学意义(F=9.474,P=0.001).异种移植组的新生血管面积均低于缝线组和碱烧伤组(P＜0.05).术后2周和4周,组织病理学检查可见异种移植组植片中的炎性细胞少于缝线组和碱烧伤组,3个组植片中均出现以CD4+T细胞为主的细胞浸润. 结论 猫-兔异种角膜移植模型较缝线和碱烧伤法制作的角膜移植模型炎症反应轻、新生血管少,角膜免疫排斥反应稳定、适度,是理想的高危角膜移植动物模型.
    배경 이상적각막이식배척동물모형시연구고위각막이식면역배척궤제적기출,구유중요적의의. 목적 비교각충건립토고위각막이식배척모형방법적림상특점,탐색합괄적각막이식배척동물모형적건립방법.방법 45지신서란백토작위각막이식수체,병안조조모방법적불동안수궤수자표법수궤분위봉선조、감소상조화이충이식조,매조15지.분별용재각막4개상한각간단봉1근5-0사선법화1 mol/LNaOH감소상법유도각막신생혈관(CNV),재건립토동충이체각막이식；령일조이묘각막위공체,건립묘-토이충각막이식모형.우제2주화제4주관찰식편적조직학정황,대3개조각막식편렬극등하관찰식편배척반응、염증화신생혈관,대식편수종정도급염증지수(IF)진행평분,근거각막혼탁、수종급신생혈관합계치계산배척지수(RI).용면역조직화학법검측CD4+T세포화CD8+T세포재식편조직중적표체. 결과 봉선조、감소상조화이충이식조분별유14、15、15지토완성천투각막이식술.술후2주,3개조IF중위수분별위0.556、0.778、0.222,차이유통계학의의(H=25.736,P=0.000),이충이식조IF치저우봉선조화감소상조,차이균유통계학의의(Z=3.841、3.993,P=0.000),봉선조IF치저우감소상조,차이유통계학의의(Z=3.568,P=0.000).술후2주,3개조RI중위수분별위2、6、3,차이유통계학의의(H=22.432,P=0.000),이충이식조RI고우봉선조이저우감소상조,차이균유통계학의의(Z=2.373,P=0.018；Z=3.936,P=0.000),봉선조RI저우감소상조,차이유통계학의의(Z=3.729,P=0.000).3개조식편존활시간분별위(17.9±2.0)、(13.4±2.4)、(15.5±2.0)d,차이유통계학의의(F=9.474,P=0.001).이충이식조적신생혈관면적균저우봉선조화감소상조(P＜0.05).술후2주화4주,조직병이학검사가견이충이식조식편중적염성세포소우봉선조화감소상조,3개조식편중균출현이CD4+T세포위주적세포침윤. 결론 묘-토이충각막이식모형교봉선화감소상법제작적각막이식모형염증반응경、신생혈관소,각막면역배척반응은정、괄도,시이상적고위각막이식동물모형.
    Background An ideal animal model is very important for the investigation of the immune mechanism of high-risk rejection corneal transplantation. Objective This study was to compare three methods of creating a high-risk corneal transplantation model in rabbits to study high-risk rejection corneal transplantation.Methods Forty-five New Zealand white rabbits were utilized and assigned randomly to three groups of different modeling methods,with 15 rabbits for each group.The high-risk corneal transplantation models were created by suturing with 5-0 silk thread in 4 quadrants,inducing alkali burn with 1 mol/L NaOH or corneal xenotransplantation.In the suturing group and alkali burning group,the rabbits received a unilateral 7.25 mm diameter corneal allograft after corneal neovascularization was induced,and in the xenotransplantation group,corneas from cats were used as donors.Rabbits were followed-up for 4 weeks in all groups.Corneal neovascular area was calculated and compared among the three groups.The amount of rejection,inflammatory index ( IF),neovascularization and histology of grafts were clinically scored to calculate the reject index (RI). Results There were 14,15 and 15 rabbits that survived the high-risk penetrating corneal transplantation,respectively,in the suturing group,alkali burning group and xenotransplantation group.Two weeks after operation,the IF scores were 0.543 ± 0.103,0.811 ± 0.054 and 0.191 ±0.087,and the RI were 2.111±0.928,7.0±0.816 and 3.182±0.751 in the suturing group,alkali burning group and xenotransplantation group,respectively,showing statistically significant differences among the three groups (x2 =25.736,22.432,P =0.000).The IF value was lower in the xenotransplantation group compared with the suturing group and alkali burning group (Z =3.841,3.993,P =0.000),and that of the suturing group was lower than the alkali burning group (Z =3.568,P =0.000).The RI value of the xenotransplantation group was significantly raised in comparison with the suturing group and declined in comparison with the alkali burning group (Z =2.373,P =0.018;Z =3.936,P =0.000),and that of the suturing group was lower than the alkali burning group (Z =3.729,P =0.000 ).The survival times of the grafts were ( 17.9±2.0 ) days,( 13.4 ±2.4) days and ( 15.5 ±2.0 ) days in these three groups with a significant difference among them ( F =9.474,P =0.001 ).The neovascularization area in the xenotransplantation group was smaller than the suturing group and alkali burning group (P＜ 0.05 ).Histological examination revealed a large number of inflammatory cells infiltration in the grafts 2 and 4 weeks after transplantation in the suturing group and alkali burning group,but less inflammatory cells were seen in the xenotransplantaion group.Immunofluorescence staining showed abundant CD4+ T positive cells in the grafts in the three groups. Conclusions The cat-rabbit corneal xenotransplantation can induce stable and moderate immune rejection.This animal model has milder inflammatory response and less corneal neovascularization than the suture and alkali burn models.This method therefore is an ideal model for high-risk corneal transplantation.
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