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
2008年
5期
996-1000
,共5页
凌士奇%张慧%林浩添%徐建刚%邝文辉
凌士奇%張慧%林浩添%徐建剛%鄺文輝
릉사기%장혜%림호첨%서건강%광문휘
高危角膜移植%免疫排斥%颈淋巴结切除
高危角膜移植%免疫排斥%頸淋巴結切除
고위각막이식%면역배척%경림파결절제
背景:颈淋巴结是角膜的引流区淋巴结.角膜移植后,抗原提呈细胞随着房水经由脉络膜巩膜外途径引流至颈淋巴结而诱发角膜免疫排斥反应.目的:观察在碱烧伤的角膜植床上行角膜移植后的免疫排斥反应现象,认识颈淋巴结切除抑制角膜移植后免疫排斥的效应.设计:随机对照动物实验.单位:中山大学中山眼科中心.材料:实验于2005-05/2007-02在中山大学中山眼科中心完成(眼科学国家重点试验室.实验室编号2006DA105054).选用SD大鼠104只, Wistar大鼠40只,均为雄性,鼠龄1～2个月,均由中山大学实验动物中心提供.实验所用的白细胞介素2 和干扰素γ ELISA试剂盒由美国Biource International公司提供.实验过程对动物的处置符合在眼科研究使用动物的ARVO声明.方法:以SD鼠为受体,Wistar鼠为供体,所有受体大鼠均行同种异体角膜移植.受体鼠被随机分为4组:正常对照组:行正常角膜移植;B组为颈淋巴结切除组:正常大鼠切除双侧颈淋巴结;碱烧伤后角膜移植组:角膜碱烧伤后21 d时行角膜移植;碱烧伤后角膜移植合并颈淋巴结切除组:角膜碱烧伤后立即行双侧颈淋巴结切除,21 d后再行角膜移植;每组20只.应用ELISA法检测角膜移植后各组植片中干扰素γ、白细胞介素-2蛋白的表达,记录角膜排斥反应发生的时间并比较各组植片平均存活时间.其余24只受体鼠用于在裂隙灯下及组织切片后显微镜下观察碱烧伤后角膜炎症及新生血管的动态变化.结果: ①病理组织学:正常角膜无炎症及新生血管.碱烧伤后3天时,角膜基质中存在着大量的炎症细胞浸润.3周末时无明显的炎症征象,但角膜新生血管达到高峰.碱烧伤后8周时,角膜新生血管已完全消退.②植片平均存活时间: 正常对照组、颈淋巴结切除组、碱烧伤后角膜移植组、碱烧伤后角膜移植合并颈淋巴结切除组分别为(10.40±1.14),(46.30±9.46),(7.00±1.58)和(15.00±3.39)d;颈淋巴结切除组较正常对照组明显延长(P < 0.05),而碱烧伤后角膜移植合并颈淋巴结切除组较碱烧伤后角膜移植组明显延长,差异具有显著性意义 (P < 0.05).③移植后角膜中干扰素γ及白细胞介素2蛋白的表达:角膜移植后颈淋巴结切除组植片中无干扰素γ及白细胞介素2蛋白的表达.在角膜移植后3,7,10,14 d,碱烧伤后角膜移植合并颈淋巴结切除组干扰素γ和白细胞介素-2的表达均明显下降,与碱烧伤后角膜移植组比较,差异有显著性意义(P < 0.05).结论:颈淋巴结切除能有效抑制正常及碱烧伤后角膜移植术后的免疫排斥反应.
揹景:頸淋巴結是角膜的引流區淋巴結.角膜移植後,抗原提呈細胞隨著房水經由脈絡膜鞏膜外途徑引流至頸淋巴結而誘髮角膜免疫排斥反應.目的:觀察在堿燒傷的角膜植床上行角膜移植後的免疫排斥反應現象,認識頸淋巴結切除抑製角膜移植後免疫排斥的效應.設計:隨機對照動物實驗.單位:中山大學中山眼科中心.材料:實驗于2005-05/2007-02在中山大學中山眼科中心完成(眼科學國傢重點試驗室.實驗室編號2006DA105054).選用SD大鼠104隻, Wistar大鼠40隻,均為雄性,鼠齡1～2箇月,均由中山大學實驗動物中心提供.實驗所用的白細胞介素2 和榦擾素γ ELISA試劑盒由美國Biource International公司提供.實驗過程對動物的處置符閤在眼科研究使用動物的ARVO聲明.方法:以SD鼠為受體,Wistar鼠為供體,所有受體大鼠均行同種異體角膜移植.受體鼠被隨機分為4組:正常對照組:行正常角膜移植;B組為頸淋巴結切除組:正常大鼠切除雙側頸淋巴結;堿燒傷後角膜移植組:角膜堿燒傷後21 d時行角膜移植;堿燒傷後角膜移植閤併頸淋巴結切除組:角膜堿燒傷後立即行雙側頸淋巴結切除,21 d後再行角膜移植;每組20隻.應用ELISA法檢測角膜移植後各組植片中榦擾素γ、白細胞介素-2蛋白的錶達,記錄角膜排斥反應髮生的時間併比較各組植片平均存活時間.其餘24隻受體鼠用于在裂隙燈下及組織切片後顯微鏡下觀察堿燒傷後角膜炎癥及新生血管的動態變化.結果: ①病理組織學:正常角膜無炎癥及新生血管.堿燒傷後3天時,角膜基質中存在著大量的炎癥細胞浸潤.3週末時無明顯的炎癥徵象,但角膜新生血管達到高峰.堿燒傷後8週時,角膜新生血管已完全消退.②植片平均存活時間: 正常對照組、頸淋巴結切除組、堿燒傷後角膜移植組、堿燒傷後角膜移植閤併頸淋巴結切除組分彆為(10.40±1.14),(46.30±9.46),(7.00±1.58)和(15.00±3.39)d;頸淋巴結切除組較正常對照組明顯延長(P < 0.05),而堿燒傷後角膜移植閤併頸淋巴結切除組較堿燒傷後角膜移植組明顯延長,差異具有顯著性意義 (P < 0.05).③移植後角膜中榦擾素γ及白細胞介素2蛋白的錶達:角膜移植後頸淋巴結切除組植片中無榦擾素γ及白細胞介素2蛋白的錶達.在角膜移植後3,7,10,14 d,堿燒傷後角膜移植閤併頸淋巴結切除組榦擾素γ和白細胞介素-2的錶達均明顯下降,與堿燒傷後角膜移植組比較,差異有顯著性意義(P < 0.05).結論:頸淋巴結切除能有效抑製正常及堿燒傷後角膜移植術後的免疫排斥反應.
배경:경림파결시각막적인류구림파결.각막이식후,항원제정세포수착방수경유맥락막공막외도경인류지경림파결이유발각막면역배척반응.목적:관찰재감소상적각막식상상행각막이식후적면역배척반응현상,인식경림파결절제억제각막이식후면역배척적효응.설계:수궤대조동물실험.단위:중산대학중산안과중심.재료:실험우2005-05/2007-02재중산대학중산안과중심완성(안과학국가중점시험실.실험실편호2006DA105054).선용SD대서104지, Wistar대서40지,균위웅성,서령1～2개월,균유중산대학실험동물중심제공.실험소용적백세포개소2 화간우소γ ELISA시제합유미국Biource International공사제공.실험과정대동물적처치부합재안과연구사용동물적ARVO성명.방법:이SD서위수체,Wistar서위공체,소유수체대서균행동충이체각막이식.수체서피수궤분위4조:정상대조조:행정상각막이식;B조위경림파결절제조:정상대서절제쌍측경림파결;감소상후각막이식조:각막감소상후21 d시행각막이식;감소상후각막이식합병경림파결절제조:각막감소상후립즉행쌍측경림파결절제,21 d후재행각막이식;매조20지.응용ELISA법검측각막이식후각조식편중간우소γ、백세포개소-2단백적표체,기록각막배척반응발생적시간병비교각조식편평균존활시간.기여24지수체서용우재렬극등하급조직절편후현미경하관찰감소상후각막염증급신생혈관적동태변화.결과: ①병리조직학:정상각막무염증급신생혈관.감소상후3천시,각막기질중존재착대량적염증세포침윤.3주말시무명현적염증정상,단각막신생혈관체도고봉.감소상후8주시,각막신생혈관이완전소퇴.②식편평균존활시간: 정상대조조、경림파결절제조、감소상후각막이식조、감소상후각막이식합병경림파결절제조분별위(10.40±1.14),(46.30±9.46),(7.00±1.58)화(15.00±3.39)d;경림파결절제조교정상대조조명현연장(P < 0.05),이감소상후각막이식합병경림파결절제조교감소상후각막이식조명현연장,차이구유현저성의의 (P < 0.05).③이식후각막중간우소γ급백세포개소2단백적표체:각막이식후경림파결절제조식편중무간우소γ급백세포개소2단백적표체.재각막이식후3,7,10,14 d,감소상후각막이식합병경림파결절제조간우소γ화백세포개소-2적표체균명현하강,여감소상후각막이식조비교,차이유현저성의의(P < 0.05).결론:경림파결절제능유효억제정상급감소상후각막이식술후적면역배척반응.
BACKGROUND: Cervical lymph nodes are draining region of cornea. It is believed that aqueous fluid goes through a minor pathway named uveoscleral drainage, which will allow passage of antigen-presenting cells (APC) directly to the draining lymph nodes and induce allograft rejection after keratoplasty.OBJECTIVE: To explore the inhibitory effects of cervical lymphadenectomy in alkali induced high-risk corneal transplantation.DESIGN: A randomized controlled animal experiment.SETTING: State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University.MATERIALS: The experiment was performed in the State Key Laboratory of Ophthalmology (No. 2006DA105054), Zhongshan Ophthalmic Center, Sun Yat-sen University from May 2005 to February 2007. 144 male animals (1-2 months old) including 104 SD rats and 40 Wistar rats were provided by the animal experimental center of Sun Yat-sen University. Sandwich enzyme-linked immunosorbent assay (ELISA) kits for interleukin-2 (IL-2) and interferon-γ (IFN-γ) were brought from BioSource International company (USA). The animal treatment in the experiment was accorded with the statement in Association for Research in Vision and Ophthalmology (ARVO) for animals.METHODS: With the SD rats as recipients, and Wistar rats as donors, all rats were subjected to corneal allografting. The recipient rats were randomly divided into 4 groups (n=20): group A (control group) which underwent corneal transplantation; group B which was subjected to bilateral cervical lymphadenectomy; group C, corneal transplantation 21 days after the alkali burn injury; group D, cervical lymphadenectomy following group C. The immune rejection of grafts was evaluated by detecting the expression of IFN-γ and IL-2 using ELISA. The time when allograft rejection occurred was recorded and mean survival time (MST) was compared among the groups. The development of corneal inflammation and new vessels was examined by slit lamp microscope and histopathological examination.MAIN OUTCOME MEASURES: ①The development of corneal inflammation after corneal alkaline burns. ②MST of rats in each group following transplant. ③The expression of IL-2 and IFN-γ in grafts of each group. RESULTS: ①Normal rat cornea was transparent without inflammation or neovascularization. There were many inflammatory cells invading to stroma on day 3 after burn. Then, the inflammation of cornea resolved gradually 3 weeks after the burn, but corneal neovascularization reached the peak at that time. Corneal blood vessels regressed completely at the end of 8 weeks after the burn. ②The MST of group A, B, C, and D was (10.40±1.14), (46.30±9.46), (7.00±1.58), and (15.00±3.39) days, respectively. Compared with the group A, the MST of group B was significantly longer (P < 0.05), and the MST of grafts in group D was also significantly longer than group C (P < 0.05). ③The expression of IFN-γ and IL-2 proteins was absent in group B. Compared with group C, the expression of IL-2 and IFN-γ proteins in group D significantly decreased on days 3, 7, 10, and 14 after keratoplasty (P < 0.05). CONCLUSION: Cervical lymphadenectomy therapy can effectively inhibit corneal allograft rejection in normal and high-risk corneal beds after alkali burn injury.
