腹泻型肠易激综合征大鼠模型的建立及痛泻要方对肠易激综合征大鼠治疗作用的实验研究

发布时间：2018-09-13 09:23

【摘要】： 目的:探讨肠易激综合征(irritable bowel syndrome)的发病机制以及痛泻要方煎剂治疗肠易激综合征内脏高敏感性大鼠的机制。方法: 1.选用清洁级新生SD大鼠,采用直肠慢性刺激加夹尾刺激法造模。于大鼠出生后第8-21d,每天给予直肠内醋酸刺激,两周后予急性应激夹尾刺激一周,在出生后第7,9,11周对这些成年后的大鼠进行直肠扩张,评估其腹部收缩反射(AWR)阈值,12周测定大鼠腹壁肌电话动,验证敏感性有无异常改变。证实造模是否成功。 2.造模成功后,将实验动物分组为中药低剂量组(A组)、中药中剂量组(B组)、中药高剂量组(C组)、西药奥替溴铵组(D组)、空白对照组(E组)、模型对照组(F组)。其中E组为正常大鼠,A、B、C、D、F组为模型大鼠。E组和F组以生理盐水按4ml/100g(40g/kg/d)灌胃,A、B、C组分别以痛泻要方煎剂按中药低剂量组0.4ml/100g(4g/kg/d)、中药中剂量组1.2ml/100g(12g/kg/d)、中药高剂量组4ml/100g(40g/kg/d)灌胃,D组西药奥替溴铵组按4ml/100g(150g/kg/d)灌胃,每天两次,连续30天。观察球囊在各组大鼠肠道内扩张引起腹部抬起和背部拱起的容量阈值以及球囊在各组大鼠肠道内不同容量下扩张腹壁收缩次数。放射免疫法测血浆P物质(SP )、结肠组织匀浆P物质(SP )、血浆降钙素基因相关肽(CGRP)、结肠组织匀浆降钙素基因相关肽(CGRP)含量。结果作统计分析。结果: 1.与新生期生理盐水刺激组(C组)和新生期醋酸刺激组(B组)相比,直肠扩张时,新生期醋酸刺激后2周加夹尾刺激组大鼠(A组)腹部抬起和背部拱起的容量阈值显著降低(P值均0.01);0.3、0.6、0.9 ml扩张容量下A组腹壁肌电活动明显增强(与B组相比,P分别均0.05;与C组相比,P分别0.01、0.05和0.05)。2.用药后,痛泻要方与斯巴敏都能使肠道致敏后所致的行为学与电生理指标恢复正常,且中药高剂量组与斯巴敏组组间无差异。而中药低剂量组和中剂量组使肠道致敏后所致的行为学与电生理指标恢复不明显,没有达到统计学意义。模型大鼠血清SP含量明显增加,痛泻要方高剂量组与西药斯巴敏组都能使模型大鼠血清SP含量明显回落,且中药高剂量组与斯巴敏组组间无差异。模型大鼠结肠粘膜SP含量明显增加,痛泻要方中、高剂量组与西药斯巴敏组都能使模型大鼠结肠粘膜SP含量明显回落,且中药中、高剂量组与斯巴敏组间无差异。痛泻要方高剂量组能显著增加血浆和结肠组织CGRP含量,痛泻要方低剂量组、中剂量组与西药组均不能。结论: 1.新生期大鼠肠道内的慢性刺激加夹尾刺激可以在成年后引起慢性内脏敏感性增高。并表明应激和肠道感染两因素共同作用时具有致肠动力紊乱的协同效果,应激使感染后的肠道易于发生动力紊乱,已被感染过的肠道对应激的反应更为强烈。而肠粘膜未见异常病理改变,符合IBS的基本特征。 2.痛泻要方煎剂能使肠道致敏后所致的行为学与电生理指标恢复正常,SP含量上升在IBS发病机制中起重要作用,IBS能导致血浆和结肠组织中CGRP含量下降。痛泻要方能降低模型大鼠血浆和结肠组织SP含量,增加CGRP含量,而斯巴敏仅能降低SP含量。本方的作用机制可能是通过降低模型大鼠血浆和结肠组织SP含量,减弱背角神经元兴奋性,从而提高内脏痛阈,消除肠道过敏。综合起来看,痛泻要方高剂量组的疗效优于斯巴敏。
[Abstract]:Objective: To investigate the pathogenesis of irritable bowel syndrome and the mechanism of Tongxieyaofang Decoction in treating visceral hypersensitivity rats with irritable bowel syndrome.
Method:
1. Neonatal SD rats of clean grade were selected and modeled by rectal chronic stimulation plus tail clamping stimulation. The rats were given intrarectal acetic acid stimulation every day for 8-21 days after birth, followed by acute stress tail clamping stimulation for one week, rectal dilatation was performed at 7, 9 and 11 weeks after birth, and the abdominal systolic reflex (AWR) threshold was assessed for 12 weeks. The abnormality of the sensitivity of the abdominal wall muscles in rats was tested.
2. After successful modeling, the experimental animals were divided into low-dose group (group A), middle-dose group (group B), high-dose group (group C), Otibromide group (group D), blank control group (group E), model control group (group F). Group E was normal rats, group A, B, C, D, F were model rats. Group E and F were given normal saline 4 ml/100 g (40 g/kg/d), and group A, B, C, D, F were given orally. Group B and group C were given Tongxie Yaofang Decoction 0.4ml/100g (4g/kg/d) in low dose group, 1.2ml/100g (12g/kg/d) in middle dose group, 4ml/100g (40g/kg/d) in high dose group, and Otibromide group D was given Otibromide Decoction 4 ml/100g (150g/kg/d) twice a day for 30 days. Volume threshold of dorsal arch and contraction times of abdominal wall dilated by balloon in different intestinal volumes of rats were measured by radioimmunoassay. The contents of substance P (SP), substance P (SP) in plasma, calcitonin gene-related peptide (CGRP) in colon tissue homogenate, and calcitonin gene-related peptide (CGRP) in plasma were measured.
Result:
1. Compared with the neonatal saline stimulation group (group C) and neonatal acetic acid stimulation group (group B), the volume threshold of abdominal elevation and dorsal arch in neonatal acetic acid stimulation group (group A) decreased significantly 2 weeks after rectal dilatation (P 0.01); the abdominal myoelectric activity in group A increased significantly (compared with group B) at 0.3, 0.6, 0.9 ml dilatation volume. Compared with C group, P was 0.05, P was 0.01, 0.05 and 0.05). 2. After treatment, both Tongxie Yaofang and Spasmone could restore the normal behavior and electrophysiological indexes caused by intestinal sensitization, and there was no difference between high dose group and Spasmone group. There was no significant difference in the recovery of physiological indexes between the two groups. The content of SP in the colon mucosa of the model rats increased significantly, and the content of SP in the high dose group of Tongxie Yaofang and the western medicine Spamin group decreased significantly. The high dose group of Tongxie Yaofang could significantly increase the content of CGRP in plasma and colon tissue, while the low dose group of Tongxie Yaofang could not.
Conclusion:
1. Chronic intestinal stimulation plus tail clip stimulation in neonatal rats can increase the sensitivity of chronic viscera in adulthood. It shows that stress and intestinal infection have synergistic effects on intestinal motility disorder. Stress makes the infected intestine prone to dynamic disorder, and the infected intestine is more responsive to stress. There was no abnormal pathological changes in intestinal mucosa, which accords with the basic characteristics of IBS.
2. Tongxie Yaofang Decoction can restore the behavioral and electrophysiological indexes induced by intestinal sensitization. The increase of SP content plays an important role in the pathogenesis of IBS. IBS can lead to the decrease of CGRP content in plasma and colon tissues. Tongxie Yaofang Decoction can decrease the SP content in plasma and colon tissues of model rats, increase the CGRP content, while Spasmone can only decrease the S content. The mechanism of action of Tongxie Yaofang may be that it can increase visceral pain threshold and eliminate intestinal allergy by lowering SP content in plasma and colon tissue of model rats and decreasing the excitability of dorsal horn neurons.
【学位授予单位】：辽宁中医药大学
【学位级别】：硕士
【学位授予年份】：2008
【分类号】：R-332;R285.5

【相似文献】