摘要
目的:角膜移植免疫排斥反应是一个多因素参与的调节过程,近年来研究表明细胞因子在角膜移植免疫中起重要作用。实验拟验证γ-干扰素、白细胞介素4及CD25在大鼠角膜移植免疫排斥反应中的作用。方法:①实验于2006-10/2007-05在南方医科大学南方医院实验动物中心完成。选用Wistar大鼠24只,SD大鼠54只,实验方法符合动物伦理学要求。②将54只SD大鼠按随机数字表法分为3组,正常组6只,角膜移植组及地塞米松组均24只。后两组制备Wistar→SD角膜移植大鼠模型,受体选择右眼手术,供体提供双眼角膜。③用裂隙灯观察移植排斥情况;利用反转录-聚合酶链反应检测植片内γ-干扰素、CD25mRNA的表达;应用酶联免疫吸附实验检测房水和血清中γ-干扰素、白细胞介素4含量;应用流式细胞术检测移植前后不同时相外周血淋巴细胞CD25的表达;术后第11天,应用免疫组织化学方法检测角膜CD4+,CD8+和CD25+T细胞的表达。结果:54只SD大鼠全部进入结果分析。①地塞米松组大鼠发生角膜移植排斥反应的时间长于角膜移植组(P=0.000)。②正常角膜植片无CD25、γ-干扰素mRNA表达,角膜移植组术后第11天角膜植片内γ-干扰素、CD25 mRNA的表达较地塞米松组增强(P<0.05)。③角膜移植组角膜移植术后6,11d血清γ-干扰素含量高于正常组(P<0.05);术后11d含量高于术后6,24d(P<0.01);地塞米松组术后6,11d低于同期角膜移植组(P<0.05)。④角膜移植组角膜移植术后6,11,24d房水γ-干扰素、白细胞介素4含量均高于正常组(P<0.05);地塞米松组术后6,11d均低于同期角膜移植组(P<0.05～0.01)。⑤地塞米松组术后6,11d外周血CD3+CD25+T/CD3+T比值低于同期角膜移植组(P<0.05)。⑥术后第11天角膜移植组植片内CD4,CD8及CD25明显表达,地塞米松组CD4,CD8及CD25表达明显减弱。结论:CD25、γ-干扰素和白细胞介素4在角膜移植免疫排斥反应过程中发挥重要的�
AIM：Corneal allograft rejection is a complicated adjusting process which multiple factors participate in. Recent researches showed that the cytokine plays an important role in corneal transplantation immunity. This study was designed to investigate the role of interferon （IFN）-γ, interleukin-4 （IL-4） and CD25 in the allograft rejection of rat penetrating keratoplasty （PKP）.
METHODS： ①Experiments were performed at the Experiment Animal Center of Nanfang Hospital of Southern Medical University from October 2006 to May 2007. A total of 24 Wistar rats and 54 SD rats were selected. Experimental procedures met the animal ethical standards. ②The 54 SD rats were randomly divided into three groups, normal control group （n=6）, corneal graft group （n=24） and dexamethasone group （n=24）. Rats in the latter two groups received Wistar→SD corneal transplantation. Receptors received right eye surgery, and donors offered cornea of both eyes. ③Rejection reaction was observed with a slit lamp microscope. Levels of IFN-γand CD25 mRNA in grafts were detected by using reverse transcription polymerase chain reaction （RT-PCR）. Aqueous humor and serum levels of IFN-γand IL-4 were measured by enzyme linked immunosorbent assay （ELISA）. The expression of CD25 in peripheral lymphocytes was measured by flow cytometry （FCM） before and after surgery. Immunohistochemical method was performed to examine corneal CD4^＋,CD8^＋ and CD25^＋T expressions at day 11 after surgery.
RESULTS： Totally 54 SD rats were included in the final results. ①The rejection time was longer in the dexamethasone group than in the corneal graft group （P =0.000）. ②There was no expression of IFN-γ and CD25 mRNA in the normal cornea. Compared to the dexamethasone group, the expressions of IFN-γ and CD25 mRNA in grafts were markedly increased in corneal graft group at day 11 after PKP （P 〈 0.05）. ③The IFN-γlevels in serum were higher in the corneal graft group than in the normal control group at day
出处
《中国组织工程研究与临床康复》
CAS
CSCD
北大核心
2008年第5期861-867,共7页
Journal of Clinical Rehabilitative Tissue Engineering Research
基金
广东省自然科学基金项目(04020438)~~
