[1]苏亚茹,白浪,刘晶,等.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[J].眼科新进展,2016,36(7):601-604.[doi:10.13389/j.cnki.rao.2016.0159]
　SU Ya-Ru,BAI Lang,LIU Jing,et al.Effects of Th cell polarization regulated by TLR2 0n immune rejection after corneal transplantation in mouse[J].Recent Advances in Ophthalmology,2016,36(7):601-604.[doi:10.13389/j.cnki.rao.2016.0159]
点击复制

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
分享到：

《眼科新进展》[ISSN:1003-5141/CN:41-1105/R]

卷:
36卷
期数:
2016年7期
页码:
601-604
栏目:
实验研究
出版日期:
2016-07-05

文章信息/Info

Title:
Effects of Th cell polarization regulated by TLR2 0n immune rejection after corneal transplantation in mouse
作者:
苏亚茹; 白浪; 刘晶; 梁伟怡; 于健; 孟婷
510000　广东省广州市，南方医科大学南方医院眼科
Author(s):
SU Ya-Ru ;  BAI Lang ;  LIU Jing ;  LIANG Wei-Yi; YU Jian; MENG Ting
Department of Ophthalmology,Nanfang Hospital of Southem Medical University,Guangzhou 510000, Guangdong Province.China
关键词:
Toll样受体2; 角膜移植; 免疫排斥; Th1; Th2; Th17
Keywords:
Toll like receptor 2 ;  corneal transplantation ;  immune rejection ;  Thl ;  Th2 ; Th17
分类号:
Ｒ７７２
DOI:
10.13389/j.cnki.rao.2016.0159
文献标志码:
A
摘要:
目的　探究Ｔｏｌｌ样受体２（Ｔｏｌｌｌｉｋｅｒｅｃｅｐｔｏｒ２，ＴＬＲ２）是否通过调节辅助性Ｔ细胞（Ｔｈｅｌｐｅｒｃｅｌｌｓ，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（ｒｅｊｅｃｔｉｎｄｅｘ，ＲＩ）进行评分；术后１４ｄ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７ｄ、１４ｄ、２１ｄ，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-１７）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。
Abstract:
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 randonuzed into the wide type mouse group and knock-out mouse group. Another 19 C57BL/6 mouse were performed isograft corneal transplantation and set as isograft group. The edema, transparency were observed twice 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 C57BL/6 and TLR2 knock-out mouse were involved. The corneal tissues were sampled at 14 days for HE staining. The mRNA expression of Thl . Th2 ,Th17 cytokine ( IFN-7,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 sigruficantly lower 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 pronunent 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-7 and IL-17 mRNA in the knock-out mouse group ( 1. 94 +0. 34 .2. 62 +0. 30) were also lower 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 immune rejection after corneal transplantation.

相似文献/References:

[1]朱勤 胡竹林 杨忠昆.恒河猴血管内皮细胞移植替代角膜内皮细胞后房水血管内皮生长因子浓度变化的观察[J].眼科新进展,2012,32(1):000.
[2]张虹 朱勤 胡竹林.BrdU示踪的恒河猴血管内皮细胞移植替代角膜内皮细胞的免疫组化染色观察[J].眼科新进展,2012,32(12):000.
[3]柯红琴 刘磊 栗静 王虎杰 徐曼.以纤维蛋白凝胶为载体培养兔角膜缘上皮细胞[J].眼科新进展,2013,33(2):000.
[4]付燕 高晓唯 李文静 杜安杰 李霞 赵旭东.未成熟树突状细胞在大鼠高危角膜移植免疫排斥反应中的作用[J].眼科新进展,2013,33(2):000.
[5]贾喆 张晓敏 赵少贞.间充质干细胞对异体角膜移植大鼠CD4+ CD25+ Foxp3+调节性T细胞水平的影响[J].眼科新进展,2012,32(4):000.
[6]付燕 高晓唯 王梦斐 杜安杰 李霞 赵旭东.未成熟树突状细胞诱导Th1/Th2偏移抑制大鼠高危角膜移植排斥反应的实验研究[J].眼科新进展,2013,33(3):000.
[7]朱勤 肖中男 孙晓梅 胡敏 刘海 胡竹林.恒河猴血管内皮细胞移植替代角膜内皮细胞后房水中离子浓度的变化[J].眼科新进展,2013,33(4):000.
[8]白华 黄一飞.Boston型人工角膜的临床并发症及其处理[J].眼科新进展,2013,33(9):000.
[9]周琨 高晓唯 蔡岩 李文静 胡裕坤 田丽丽 付燕. CCR7基因修饰的未成熟树突状细胞在大鼠高危角膜移植免疫排斥反应中的作用[J].眼科新进展,2014,34(1):017.
[10]李雪 王海燕 郝永娜 赵少贞 刘文广 李亚楠 边领斋 粘红 魏瑞华. 负载可递送内皮抑制素基因的胶原基复合角膜替代物在兔角膜移植中的研究[J].眼科新进展,2014,34(2):108.
[11]苏亚茹,白浪,汤明芳,等.敲除TLR2基因对同种异体小鼠角膜移植排斥反应的影响[J].眼科新进展,2016,36(11):1006.[doi:10.13389/j.cnki.rao.2016.0269]
　SU Ya-Ru,BAI Lang,TANG Ming-Fang,et al.Variation of immune response after allogeneic corneal transplantation in TLR2 knock-out mice[J].Recent Advances in Ophthalmology,2016,36(7):1006.[doi:10.13389/j.cnki.rao.2016.0269]

备注/Memo

备注/Memo:
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）
更新日期/Last Update: 2016-07-12