TLRs在大鼠穿透角膜移植术后排斥反应中的作用  

Study on effect of TLRs in rejection reaction after penetrating keratoplasty

作　　者：白浪[1] 陆晓和[2] 于健[1] 钟彦彦[2] 武海军[2] 熊柯[1] 

机构地区：[1]南方医科大学附属南方医院眼科,广州510515 [2]南方医科大学附属珠江医院眼科,广州510282

出　　处：《眼科研究》2010年第6期491-495,共5页Chinese Ophthalmic Research

基　　金：南方医院院长基金项目(2008B017)

摘　　要：目的动态检测大鼠角膜组织TLR2、TLR3、TLR4mRNA的表达,研究TLR2、TLR3、TLR4在穿透角膜移植术后免疫排斥反应中的作用。方法采用SPF级雌性SD大鼠108只为受体,SPF级Wistar大鼠36只为供体,另取SPF级雌性SD大鼠6只12只眼为正常对照组。受体大鼠分为3组:自体角膜移植组(A)、同种异体角膜移植组(B)和同种异体角膜移植后糖皮质激素应用组(C),A、B、C组各36只大鼠接受穿透角膜移植术,术后裂隙灯下观察角膜透明度、新生血管情况并采用排斥反应指数(RI)进行评分。分别于术后第5、7、9天获取角膜标本行组织病理学检查和荧光实时定量PCR(real-timePCR)检测,动态观察角膜组织中TLR2、TLR3、TLR4mRNA的表达。结果术后随时间变化,A、B、C组大鼠角膜植片均出现不同程度的水肿、混浊、新生血管生长。B组大鼠角膜RI平均在术后7d符合排斥反应的诊断标准并经病理学检查证实。A组和C组大鼠角膜的RI计分未达到排斥反应诊断标准。正常大鼠角膜可见TLR2、TLR3、TLR4mRNA表达;术后9dB组大鼠角膜TLR2mRNA的表达较A组显著增加,差异有统计学意义(P<0.05);TLR3mRNA和TLR4mRNA在B组各时间点间表达的差异倍数比较均无统计学意义(P=0.30;P=0.32),但组间比较差异均有统计学意义(P<0.05)。结论 TLR2可能作为天然免疫识别受体识别移植抗原,参与角膜移植术后的免疫排斥反应。BackgroundToll-like receptors（TLRs）can initiate the nature immunity and further induce the acquired immunity.However,whether TLRs can recognize foreign to mediate the immunity rejection following the organ or tissue transplantation is seldom reported.ObjectiveThe present study was to explore the role of TLR2,TLR3 and TLR4 in graft rejection following penetrating keratoplasty.MethodsOne hundred and eight SPF female SD rats were as receipts,and 36 SPF Wistar rats served as donors in this study.The homograft of cornea was performed on 36 receipts rats.Allografts of cornea were curried out in 72 receipts,and hormone was topically administered in 36 receipts undergone allograft rats.The another 6 matched normal rats served as normal control group.The transparency and neovascularization of cornea were examined by slit-lamp microscope and the rejection index of graft was calculated based on the inflammatory score of graft.Corneas were exsected for histopathological examination,and real-time PCR（RT-PCR）was employed to determine the expressions of TLR2,TLR3 and TLR4 mRNA in grafts in the fifth day,the seventh day and the ninth day after operation.The usage of animals followed the Statement of ARVO.ResultsThe thickness of graft and growth of new blood vessel were seen in the rats of homograft group and hormone group but no rejection occurred till the ninth day after operation.However,in allograft group,the graft rejection appeared in the seventh day,showing the edema,opacity and neovascularization of cornea.The structural disorder of graft and a lots of inflammatory cells were seen in the rejected grafts in allograft group,and mild inflammatory reaction was found in the grafts of homograft group and hormone group.The expressions of TLR2 mRNA in the grafts of allograft group were significantly higher in 9 days compared with 5-7 days after operation（P0.05）.The intergroup comparison showed the evidently changes in different groups in various time points（P0.05）,but obviously difference was exhibited in the expr

关 键 词：TOLL样受体家族 穿透角膜移植 排斥反应 实时定量PCR 

分 类 号：R779.65[医药卫生—眼科]