[1]郑艳华,白浪,梁伟怡,等. TLR2/MyD88信号系统在大鼠角膜移植术后排斥反应中的作用[J].眼科新进展,2015,35(6):506-510.[doi:10.13389/j.cnki.rao.2015.0138]
　ZHENG Yan-Hua,BAI Lang,LIANG Wei-Yi,et al. Role of TLR2/MyD88 signaling system in immune rejection after corneal transplantation in rat[J].Recent Advances in Ophthalmology,2015,35(6):506-510.[doi:10.13389/j.cnki.rao.2015.0138]
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 TLR2/MyD88信号系统在大鼠角膜移植术后排斥反应中的作用
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《眼科新进展》[ISSN:1003-5141/CN:41-1105/R]

卷:
35卷
期数:
2015年6期
页码:
506-510
栏目:
实验研究
出版日期:
2015-06-05

文章信息/Info

Title:
 Role of TLR2/MyD88 signaling system in immune rejection after corneal transplantation in rat
作者:
 郑艳华; 白浪; 梁伟怡; 于健; 杨娟; 谭青
 510515　广东省广州市，南方医科大学研究生院
Author(s):
 ZHENG Yan-Hua;  BAI Lang;  LIANG Wei-Yi;  YU Jian;  YANG Juan ;  TAN Qing
 Graduate School of Southern Medical University, Guangzhou 510515 , Guangdong Province , China
关键词:
 ; TLR2单克隆抗体; TLR2/MyD88信号系统; 角膜移植; 免疫排斥
Keywords:
 ; TLR2 monoclonal antibody;  TLR2/MyD88 signaling system;  corneal transplantation ;  immune rejection
DOI:
10.13389/j.cnki.rao.2015.0138
文献标志码:
A
摘要:
 目的　探讨Ｔｏｌｌ样受体２（Ｔｏｌｌｌｉｋｅｒｅｃｅｐｔｏｒ，ＴＬＲ２）／ＭｙＤ８８信号系统在大鼠角膜移植术后排斥反应中的作用。方法　取１４只ＳＤ大鼠为供体，２８只Ｗｉｓｔａｒ大鼠为受体，建立２８个同种异体穿透性角膜移植模型。建模后分为同种异体角膜移植组１４只（６只用于术后观察排斥情况），同种异体角膜移植ＴＬＲ２单克隆抗体组１４只（６只用于术后观察排斥情况）。另选１６只Ｗｉｓｔａｒ大鼠，分为同种自体角膜移植组８只，正常对照组８只。３个手术组大鼠分别行穿透性角膜移植术。术后ＴＬＲ２单克隆抗体组给予０．５ｇ?Ｌ－１ＴＬＲ２单克隆抗体，分别于术后０ｄ、２ｄ、４ｄ、６ｄ、８ｄ分５次经球结膜下注射给药。正常对照组、同种自体角膜移植组及同种异体角膜移植组给等量生理盐水。术后每天于裂隙灯下观察角膜透明度、新生血管，并用排斥反应指数评分。第９天取材，行ＨＥ、免疫组织化学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角膜植片均出现不同程度的水肿、混浊和新生血管生长，以同种异体角膜移植组最为明显。ＨＥ染色结果显示在同种异体角膜移植组，角膜基质层出现水肿、大量炎症细胞浸润和新生血管，而在同种自体角膜移植组和ＴＬＲ２单克隆抗体组中炎症反应较轻。免疫组织化学检测结果：ＴＬＲ２和ＭｙＤ８８分子在正常对照组、同种自体角膜移植组及ＴＬＲ２单克隆抗体组的角膜上皮中均有微量表达，同种异体角膜移植组角膜上皮、基质细胞中ＴＬＲ２和ＭｙＤ８８分子的表达明显增多，尤以基质层明显。实时荧光定量ＰＣＲ结果显示同种异体角膜移植组角膜组织ＴＬＲ２ｍＲＮＡ、ＭｙＤ８８ｍＲＮＡ的表达较同种自体角膜移植组（Ｐ＝０．０００、０．００４）和正常对照组（Ｐ＝０.０００、０．０００）显著增加，两者的表达亦较ＴＬＲ２单克隆抗体组显著增加（Ｐ＝０．０００、０．００３）。结论　ＴＬＲ２单克隆抗体抑制ＴＬＲ２及其下游信号分子ＭｙＤ８８的表达；ＴＬＲ２／ＭｙＤ８８信号系统参与了角膜移植术后排斥反应，影响了角膜植片的转归。
Abstract:
 Objective To explore the role of Toll like receptor (TLR2)/MyD88 signaling system in immune rejection after penetrating keratoplasty ( PKP) in rats. Methods Fourteen SD rats were chosen as the donators.and 28 Wistar rats as receptor ,28 allograft corneal transplantation models were established. The models were divided int0 14 allograft corneal transplantation group (6 cases were used to observe the postoperative rejection) and 14 allograft corneal transplantation treated with TLR2 monoclonal antibody group ( 6 cases were used to observe the postoperative rejection) . Another 16 Wistar rats were chosen and divided into isograft corneal transplantation group (8 cases) and normal control group (8 cases). PKP was performed in three corneal transplantation groups. The isograft and allograft groups were treated with saline .while TLR2 monoclonal antibody group with equal numbers of 0. 5 g . L-l TLR2 monoclonal antibody , they were injected at o day ,2 days ,4 days,6 days and 8 days from the bulbar conjunctiva, respectively. The corneal opacity and neovascularization were observed by slit-lamp microscope and scored according to the rejection index. with normal comea serving as the control. The corneal tissues were sampled at 9 days after the transplantation for HE and immunohistochemistry stairung. The expression of TLR2 and MyD88 mRNA were detected by qPCR. Results With the passage of time , edema, opacities and neovascularization of the corneal graft occurred after the operation in three corneal transplantation groups , especially in allograft group. HE stairung showed that severe corneal edema , a lots of inflammatory cells infiltration and new vessels in stroma were seen in allograft group, while mild inflammatory response was found in isograft group and TLR2 monoclonal antibody group. Immunohistochemistry staining result display that TLR2 and MyD88 were weakly detected in normal group ,isograft group and TLR2 monoclonal antibody group , they were increased in epithelium and stroma in allograft group , particularly in stroma. The expression of TLR2 mRNA and MyD88 mRNA in allograft group were significantly higher than those in isograft group (P = 0. 000 .0. 004) and normal control group(P = 0. 000 , 0. 000 ) , in addition. the expression in TLR2 monoclonal antibody group were significantly decreased in comparison with those in allograft group (P = 0. 000 ,0. 003 ) . Conclusion TLR2 monoclonal antibody inhibits the expression of TLR2 and its downstream signal MyD88 ;TLR2/MyD88 signaling system is involved in corneal transplantation immune rejection, and effect the outcome of corneal graft.

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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匹配基金资助（编号：Ｇ２０１２０２）
更新日期/Last Update: 2015-06-01