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背景:核因子κB在转录水平调控着许多细胞因子、黏附因子的表达,在角膜移植排斥反应中可能起着中心调控作用。目的:观察核因子κB和细胞间黏附分子1、血管内皮生长因子在角膜植片中的动态表达规律以及环孢霉素A的干预作用。设计、时间及地点:随机对照动物实验,于2005-01/07在解放军第一军医大学珠江医院眼科完成。材料:选用健康清洁级SD大鼠40只及Wistar大鼠50只。随机分为3组:同基因移植组Wistar大鼠10只为供者,Wistar大鼠20只为受者;同种异体移植组Wistar大鼠10只为供者,SD大鼠20只为受者;同种异体移植+环孢霉素A治疗组Wistar大鼠10只为供者,SD大鼠20只为受者。方法:建立大鼠角膜移植模型,移植后所有受者术眼结膜下注射庆大霉素,隔日1次,每次2000U,共用3次;2.5g/L氯霉素眼液滴眼,2次/d,每次2滴,连续用18d。5g/L托品酰胺眼液滴眼,1次/d,每次2滴,连用1周。环孢霉素A治疗组移植后第1天开始用10g/L环孢霉素A眼液滴眼,3次/d,每次2滴,连续用18d。主要观察指标:各组移植后3,7,12,18d测定角膜移植排斥反应指数评分,并于各时间点观察角膜植片病理学变化,检测核因子κB与细胞间黏附分子1、血管内皮生长因子的表达。结果:在观察期18d内,同基因组未出现排斥反应,同种异体移植组在各时间点的排斥反应指数高于同基因移植组(P<0.05),而同种异体移植+环孢霉素A治疗组排斥反应指数低于同种异体移植组(P<0.05)。免疫组织化学结果显示:核因子κB与细胞间黏附分子1、血管内皮生长因子分布角膜上皮层、基质层及新生血管内皮细胞。各时间点,同种异体移植组角膜中核因子κB与细胞间黏附分子1、血管内皮生长因子的表达高于同基因移植组(P<0.05),而低于同种异体移植+环孢霉素A治疗组(P<0.05)。结论:环孢霉素A通过减弱核因子κB核转位和发挥活性,抑制受其调控的多种细胞因子、黏附分子等移植排斥相关因子的表达,从而抑制角膜移植排斥反应的发生发展。
BACKGROUND: Nuclear factor κB regulates the expression of many cytokines and adhesion molecules at the transcriptional level. It may play a central role in the rejection of corneal allografts. Objective: To observe the dynamic expression of nuclear factor kappa B and intercellular adhesion molecule-1 and vascular endothelial growth factor in corneal graft and the intervention of cyclosporin A. DESIGN, TIME AND SETTING: The randomized controlled animal experiment was performed at the Department of Ophthalmology, Zhujiang Hospital, First Military Medical University, People’s Liberation Army from January to July 2005. MATERIALS: Forty healthy SD rats and 50 Wistar rats were selected. The rats were randomly divided into 3 groups: 10 Wistar rats in the same gene transplantation group and 20 Wistar rats as recipients; 10 Wistar rats in allograft group as donors and 20 SD rats as recipients ; Allograft + cyclosporin A treatment group Wistar rats 10 as donors, 20 SD rats as recipients. Methods: Rat model of corneal transplantation was established. Gentamycin was injected subconjunctivally into the eyes of all recipients after transplanted, once every other day for 2U and 3 times. 2.5g / L chloramphenicol eye drops and 2 times / d, 2 drops each time, continuous use 18d. 5g / L tropicamide eye drops, 1 / d, each 2 drops, once every 1 week. Cyclosporine A treatment group began to fall on the first day after transplantation with 10g / L cyclosporin A eye drops, 3 times / d, each 2 drops for 18 consecutive days. MAIN OUTCOME MEASURES: Corneal graft rejection index was measured at 3, 7, 12 and 18 days after transplantation. Pathological changes of corneal graft were observed at different time points. Nuclear factor κB, intercellular adhesion molecule 1, Factor expression. Results: In the 18d observation period, there was no rejection in the same genome. The allograft rejection index at all time points was higher than that in the allograft group (P <0.05), while the allograft + cyclosporine The rejection index of A group was lower than that of allograft group (P <0.05). Immunohistochemistry results showed that NF - kappaB was associated with ICAM - 1, VEGF distribution in the corneal epithelium, stroma and neovascular endothelial cells. At each time point, the expression of nuclear factor kappa B, intercellular adhesion molecule 1 and vascular endothelial growth factor in the cornea of allograft group was higher than that of the same gene transplantation group (P <0.05), but lower than that of allograft + cyclosporine A treatment group (P <0.05). Conclusion: Cyclosporine A can inhibit the occurrence and development of corneal graft rejection by attenuating the nuclear translocation of nuclear factor κB and its activity, inhibiting the expression of various cytokines and adhesion molecules regulated by it.