大鼠Crohn病模型NF—κB的动态变化及其对ICAM-1表达的调控  

Change of Nuclear Factor-kappa B and Regulation to the Expression of Intercellular Adhesion Molecule-1 in Rat Model of Crohn＇s Disease

作　　者：龙友明[1] 崔淑兰[1] 谢文瑞[2] 

机构地区：[1]广东药学院临床医学院,广东广州510080 [2]广东药学院附属第一医院内科

出　　处：《中国肛肠病杂志》2008年第7期9-13,共5页Chinese Journal of Coloproctology

基　　金：广东省卫生厅课题（项目编号：A2005325）

摘　　要：为探讨核转录因子-Kappa B（NF-κB）DNA结合活性在Crohn病模型的结肠组织中不同时间点的变化，以及其对细胞间黏附分子-1（ICAM-1）表达的调控作用，将36只SD大鼠平均分为6组，生理盐水对照组和5个模型组。大鼠结肠内灌注5％三硝基苯磺酸（TNBS）与无水乙醇混合液建立动物模型，于灌注后1、3、7、14、21d检测结肠病理变化；采用凝胶迁移滞留试验（EMSA）的方法测定病变结肠组织中NF-κB DNA结合活性在模型组不同时间点的变化；原位杂交及免疫组化法测定ICAM-1在上述时间点的表达。结果显示，模型组致炎后动物肠炎表现3～5d达高峰；病理组织学出现肠炎表现逐日加重，21d溃疡开始愈合，炎症慢性化，腺体增生。生理盐水组基本上无炎症性反应。肠内灌注TNBS无水乙醇后1d，结肠NF-κB明显活化，14d达到最高峰；ICAM-1 mRNA与蛋白表达在1d开始有所增加，21d达高峰。NF-κBDNA结合活性ICAM-1表达呈高度正相关。结果表明，5％TNBS加无水乙醇灌肠制作的模型与人类Crohn病相似，模型的病变结肠组织NF-κB DNA结合活性异常并持续升高，NF-κB参与TNBS无水乙醇诱导的结肠炎症的发生和发展；ICAM-1在模型的不同时间点表达增加，并受到NF-CB调控。The objective of the experiment was to detect the change of DNA-binding activity of nuclear factor-kappa B（NF-κB） at various time and the regulation to the expression of intercellular adhesion molecule- 1（ICAM-1）. Thirty six SD rats were evenly divided into 6 groups, group 1 received colonic perfusion of normal saline（the controlled group） ,and the rest 5 groups received colonic perfusion of the mixture of 5% trinitrobenzene sulfonic acid（TNBS） and absolute ethanol（the model groups）. The pathological changes of the rat colon in the 5 groups were measured on the 1st, 3rd, 7th, 14th and 21st day after perfusion. Electrophoretic mobility shift assay（EMSA） was applied in the mensurement of the change of DNA-binding activity of NF-κB at the different days;in situ hybridization（ISH） and immunohistochemistry（IH） were applied in the detection of the expression of ICAM-1 at the different days. As results, in all the 5 groups of rat models, colitis and the most severe symptoms emerged on day 3 - 5, the pathohistological changes of the colitis became aggravation then,and on day 21 the colonic ulcer began to heal,the inflammation became chro- nicity and the body of gland hyperplasia. While there was no inflammatory reaction in the controlled group. Results also showed that in the 5 model groups,colonic NF-κB began to be activated at day 1 and reached the highest level on day 14 after TNBS and absolute ethanol perfusion;that levels of ICAM-1 mRNA and protein began to increase on day 1 ,and to reach the peak on day 21 ;and that the DNA-binding activity of NF-κB and the ICAM-1 expression were positively correlated. It is concluded that the colonic lesion in rat model is similar to crohffs disease in human,the abnormally increased DNA-binding activity of NF-κB suggests the involvement in the occurrence and development of colonic inflammation. The ICAM-1 expression is also regulated by NF-κB during colonic inflammation.

关 键 词：大鼠 三硝基苯磺酸 克罗恩病 动物模型 核因子 细胞问黏附分子 

分 类 号：R644[医药卫生—外科学] R574.62[医药卫生—临床医学]