肠易激综合征大鼠结肠组织的蛋白质组学研究  

Proteomic expression analysis of colonic mucosa in a rat model of irritable bowel syndrome

作　　者：丁莺[1] 吕宾[1] 孟立娜[1] 范一宏[1] 郭赟[1] 沈雁[1] 陈硕[2] 

机构地区：[1]浙江中医药大学附属第一医院消化内科,杭州310006 [2]北京蛋白质组研究中心

出　　处：《中华医学杂志》2010年第8期564-569,共6页National Medical Journal of China

摘　　要：目的探讨肠易激综合征（IBS）的发病机制及差异蛋白质之间的网络调控关系。方法16只雌性清洁级SD大鼠按随机数字表法分人IBS组和空白对照组，每组8只。以樟脑丸特殊气味作为条件刺激、结直肠扩张结合经典的肢体束缚作为非条件刺激建立大鼠IBS模型。分别提取IBS组和空白对照组大鼠结肠黏膜总蛋白，应用差异荧光双向凝胶电泳技术进行差异蛋白质筛选，对筛选出的差异蛋白质应用基质辅助激光解吸电离飞行时间质谱技术进行鉴定，应用Gene Ontology软件对差异蛋白质进行功能分类。以蛋白质印迹法对个别有价值的差异蛋白质进行验证。结果在1396个蛋白点中筛选出19个差异蛋白点，鉴定出13个特异蛋白质，其中8个蛋白质在IBS大鼠结肠黏膜组织中表达上调，5个下调。8个表达上调的蛋白质分别是角蛋白8、蛋白二硫化物异构酶A3（PDIA3）、过氧化物氧化还原酶6、半胱氨酸组织蛋白酶S、异质性胞核核糖核蛋白F、真核细胞翻译启始因子5A、羰基还原酶1和乙二醛还原酶Ⅰ，5个表达下调的蛋白分别是α-烯醇化酶、Transgelin蛋白、丝氨酸蛋白酶抑制因子B5、心脏α-肌动蛋白1和40S核糖体蛋白SA。蛋白质印迹检测结果证实，PDIA3在IBS大鼠结肠黏膜组织中高表达，与蛋白质组学研究结果一致。结论IBS大鼠结肠黏膜组织中与肠道免疫、炎症、神经相关的某些蛋白分子的表达发生改变，提示IBS的发生与肠道免疫、炎症、神经内分泌网络调控变化有关。Objective To explore the pathogenesis of irritable bowel syndrome （IBS） and the regulation of the network relationship between differential proteins. Methods Sixteen female SD rats of clean grade were randomly divided into IBS group （ n = 8） and control group （ n = 8）. A rat model of IBS was established by a special odor of mothball as a conditional stimulation and colorectal distension plus the classic conditions of physical restraint as a non-conditional stimulation in turn. The total proteins were extracted from colon mueosa of two different rat groups. After preparation of total proteins, solubilized proteins were separated by two-dimensional differential gel electrophoresis. Those differential protein spots were identified by matrix-assisted laser desorption/ionization time of flight mass spectrometry （ MALDI-TOF MS）. The identified proteins were classified through an online web gene ontology tool. Some of the valuable differently expressed proteins were chosen for further verification by Western blotting. Results Intensity changes of 19 spots were detected among 1396 protein spots. A total of 13 different proteins were identified by MALDI-TOF MS successfully; eight of these were up-regulated and five down-regulated in colon mucosa of IBS rat. The eight over-expressed proteins were cytokeratin 8, protein disulfide isomerase A3 （ PDIA3）, peroxiredoxin-6, cathepsin S, heterogeneous nuclear ribonucleoprotein F, eukaryotic translation initiation factor, carbonyl reductase 1, glyoxalase I; five under-expressed proteins were alpha-enolase, transgelin, serpinBS, cardiac α-actin 1 and 40S ribosomal protein SA. The results of Western blotting confirmed that PDIA3 was indeed over-expressed in colonic mucosa of IBS rat. Conclusions Some proteins related to immunity of intestinal tract, inflammation and nerve are differently expressed in colonic mucosa of IBS rat. It is suggested that immunity, inflammation and neuro-endocrine network in gut are associated with the pathogenesis of IBS.

关 键 词：肠易激综合征 蛋白质组学 结肠 大鼠 

分 类 号：R574[医药卫生—消化系统]