Toll样受体2调节Th细胞极化对小鼠角膜移植排斥反应发生的影响  

Effects of Th cell polarization regulated by TLR2 on immune rejection after corneal transplantation in mouse

作　　者：苏亚茹 白浪[1] 刘晶[1] 梁伟怡[1] 于健[1] 孟婷[1] 

机构地区：[1]南方医科大学南方医院眼科,广东省广州市510000

出　　处：《眼科新进展》2016年第7期601-604,共4页Recent Advances in Ophthalmology

基　　金：国家自然科学基金资助(编号:81170887);南方医科大学南方医院横向课题基金资助(编号:G201202)~~

摘　　要：目的探究Toll样受体2(Toll like receptor 2,TLR2)是否通过调节辅助性T细胞(T helper cells,Th)极化影响角膜移植排斥反应的发生。方法取30只C57BL/6小鼠和30只TLR2基因敲除小鼠为受体,BALB/c小鼠为供体,在受体小鼠右眼行同种异体角膜移植术,分别为野生组和基因敲除组;取19只C57BL/6小鼠右眼行自体角膜移植术,为自体组。术后每周两次裂隙灯下观察植片水肿程度和透明度,参照Sonoda评分标准对排斥指数(reject index,RI)进行评分;术后14 d收集角膜。分别取9只C57BL/6小鼠和TLR2基因敲除小鼠作为野生对照组和基因敲除对照组,行常规HE染色和实时荧光定量PCR检测。结果术后随时间变化各手术组植片水肿和混浊程度不一,以野生组最为严重。角膜RI评分显示术后7 d、14 d、21 d,基因敲除组(0.80±0.83、0.90±0.55、1.35±0.99)低于野生组(1.55±0.94、2.25±0.97、2.55±1.19),差异均有统计学意义(P=0.008、0.000、0.000)。HE染色显示,野生组角膜基质层出现水肿和大量炎性细胞;而基因敲除组和自体组炎症反应较轻。PCR检测显示,基因敲除组角膜Th1和Th17细胞因子(IFN-γ和IL^(-1)7)mRNA表达(1.94±0.34、2.62±0.30)比野生组(4.27±0.37、3.99±0.40)低,差异均有统计学意义(P=0.000、0.001);Th2细胞因子(IL-4)三组差异无统计学意义(P>0.05)。结论敲除TLR2可能通过抑制Th向Th1和Th17极化,降低角膜移植排斥率。Objective To explore whether the immune rejection is adjusted by the polarization of Th cell regulated by Toll like receptor 2（ TLR2） after penetrating keratoplasty（ PK） in mouse. Methods Allograft corneal transplantation was performed on the right eyes of 30 C57BL/6 mouse and 30 TLR2 knock-out mouse to establish PK models,with BALB/c mouse as donors,which were randomized into the wide type mouse group and knock-out mouse group.Another 19 C57 BL/6 mouse were performed isograft corneal transplantation and set as isograft group. The edema,transparency were observed tw ice per week under slit lamp microscope after surgery,and rejection index（ RI） was scored according to the criteria of Sonoda. As a negative control,normal grafts from wild-type C57 BL/6 and TLR2 knock-out mouse were involved. The corneal tissues were sampled at 14 days for HE staining. The mRNA expression of Th1,Th2,Th17 cytokine（ IFN-γ,IL-4,IL-17） mRNA were detected by qPCR. Results With the passage of time,edema and opacification of corneal graft occurred after the operation in three corneal transplantation groups,especially in the wide type mouse group. The RI scores were significantly low er in the knock-out mouse group（ 0. 80 ± 0. 83,0. 90 ± 0. 55,1. 35 ± 0. 99） than those in the wide type mouse group（ 1. 55 ± 0. 94,2. 25 ± 0. 97,2. 55 ± 1. 19） at 7 days,14 days,21 days after operation（ P = 0. 008,0. 000,0. 000）. The HE staining revealed that corneal stromal edema,infiltration of inflammatory cells were more prominent at 14 days after operation in the wide type mouse group than those in the knock-out mouse group or isograft group. In addition,the expressions of IFN-γ and IL-17 mRNA in the knock-out mouse group（ 1. 94 ± 0. 34,2. 62 ± 0.30） were also low er than those in the knock-out mouse group（ 4. 27 ± 0. 37,3. 99 ± 0. 40）（ P= 0. 000,0. 001）,but not the expression of Th2 cytokine mRNA（ P 〉 0. 05）. Conclusion Inhibiting TLR2 gene to modulate the polarization of Th cell may decrease the im

关 键 词：TOLL样受体2 角膜移植 免疫排斥 TH1 TH2 Th17 

分 类 号：R772[医药卫生—眼科]