痛泻要方对内脏高敏感大鼠结肠黏膜蛋白质表达谱的影响

摘要

背景:肠易激综合征(IBS)是一种常见的功能性肠道疾病,近年研究发现其发生可能与分子异常改变有关.蛋白质组学的研究方法已用于消化系统肿瘤疾病,但较少用于功能性胃肠病的研究.目的:应用蛋白质组学技术观察痛泻要方对内脏高敏感大鼠结肠黏膜蛋白质表达谱的影响.方法:24只雌性Sprague-Dawley大鼠随机分为空白对照组、模型对照组和治疗组.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内脏高敏感模型.治疗组以痛泻要方煎剂灌胃4周.实验第45 d,处死所有大鼠,提取结肠黏膜总蛋白.应用荧光差异双向凝胶电泳技术筛选差异蛋白质,基质辅助激光解吸电离飞行时间质谱技术鉴定差异蛋白质.结果:与模型对照组相比,治疗组内脏敏感性明显降低(P＜0.01).共有61个蛋白点在模型对照组和治疗组中存在表达差异,其中23个蛋白点在治疗组中的表达上调,38个蛋白点表达下调.从5个明显差异表达的蛋白点中鉴定出3个特异蛋白质,表达上调的蛋白质为Transgelin蛋白和乙醛脱氢酶2,表达下调的蛋白质为角蛋白8.结论:痛泻要方可下调某些炎症相关蛋白的表达,这可能是其降低大鼠内脏敏感性的机制之一.%Background: Irritable bowel syndrome (IBS) is a common functional bowel disorder. Recent studies have shown that molecular changes may be involved in the pathogenesis of IBS. Proteomics has been applied to study digestive system tumors, however, it is seldomly used in study of functional gastrointestinal disorders. Aims: To investigate the effect of Tongxieyaofang on colonic mucosal protein expression profile in rats with visceral hypersensitivity by proteomics technology. Methods: Twenty-four female Sprague-Dawley rats were randomly divided into blank control group, model control group and treatment group. Visceral hypersensitivity model was established by special odor of mothball as conditional stimulation and colorectal distension combined with classic physical restraint as non-conditional stimulation. Rats in treatment group were intragastrically treated with Tongxieyaofang for 4 weeks. All the rats were sacrificed on the 45th day. Total proteins were extracted from colonic mucosa. Differential protein spots were screened by fluorescence two-dimensional differential gel electrophoresis, and were identified by matrix-assisted laser desorption/ionization time of flight mass spectrometry. Results: Visceral sensitivity in the treatment group was lower than that in the model control group (P<0. 01). Sixty-one protein spots were significantly differentially expressed between model control group and treatment group, 23 spots of these were up-regulated while 38 spots were down-regulated in treatment group. Three specific proteins were successfully identified from 5 protein spots with marked changes. The two up-expressed proteins were Transgelin and acetaldehyde dehydrogenase 2, the down-expressed protein was cytokeratin 8. Conclusions: Tongxieyaofang can down-regulate the expression of inflammation-related proteins, which may be one of the mechanisms of Tongxieyaofang in decreasing visceral hypersensitivity in rats.