【作者】 罗琦；

【导师】 周福生；

【作者基本信息】 广州中医药大学 ， 中西医结合临床， 2012， 博士

【摘要】 肠易激综合征(IBS)是一种常见的以腹痛／腹部不适伴排便习惯改变为特征的功能性肠病,缺乏形态学和生化学改变的生物学标志。IBS的发病机制迄今仍不明确,现在普遍认为是多种因素综合作用的结果,其中心理、精神、社会和环境等因素在发病过程中起重要作用,同时受神经系统、免疫系统和内分泌系统的制约,导致肠道平滑肌运动障碍和内脏感觉异常。在此基础上,当机体受到某些诱因刺激时就会引起肠道易激和过激的功能反应,导致一系列临床症状。其症状发生的直接机制是基于肠道高敏感性和高反应性基础上的肠道运动功能异常。本文通过文献回顾了IBS的发病、致病机理、诊断治疗及肾上腺皮质激素释放因子(Corticotropin-releasing hormone, CRH)通路和肥大细胞在IBS致病作用的研究。目的本课题旨在通过IBS大鼠模型实验研究,探讨肠易激综合征的发生是否与肾上腺皮质激素释放因子(Corticotropin-releasing hormone, CRH)通路和肥大细胞相关。从而加深我们对肠易激综合征的发病机制的认识,并有助于解释中药康泰复方治疗肠易激综合征的作用途径及机制,为心胃相关理论治疗肠易激综合征的现代化生物学基础提供科学根据及理论支持。方法将育龄7天的新生SD大鼠共110只随机分出正常对照组10只,其中100只利用幼鼠母子分离加直肠刺激法复制大鼠IBS内脏高敏感性模型,造模成功后按体重随机分出模型组、中药高、中、低剂量组,和西药马来酸曲美布汀对照组各20只。给药两周后运用放射免疫法检测大鼠模型下丘脑、脑干CRH水平,免疫组化法检测IBS内脏高敏感性大鼠肠粘膜肥大细胞数目、活化状态(c-fos基因表达情况)及中药康泰复方对其影响；并与模型组及马来酸曲美布汀组进行比较。结果1正常大鼠与造模大鼠不同时期体重变化用幼鼠母子分离加直肠刺激法复制内脏高敏感性IBS大鼠与正常对照组大鼠体重变化无显著性差异(P>0.05),说明本法造模不会影响大鼠的正常状态。2大鼠在清醒状态下,逐渐加压扩张肠道,观察大鼠腹部抬起与背部拱起的血压计的压力平均值,采用幼鼠母子分离加直肠刺激法复制内脏高敏感性IBS大鼠模型,在大鼠出生后第六周其内脏高敏感性显著升高(P<0.05),说明造模成功。3、IBS内脏高敏感性大鼠经康泰胶囊治疗2周后,用药组大鼠内脏高敏感性明显降低(P<0.05),且中、高剂量治疗组已恢复到接近正常水平(P>0.05)。提示康泰胶囊对IBS模型大鼠内脏高敏感性有改善作用。4、康泰胶囊治疗后,用幼鼠母子分离加直肠刺激法复制内脏高敏感性IBS大鼠与正常对照组大鼠体重、活动、毛发光泽无显著性差异(P>0.05),说明康泰胶囊是安全的,不存在明显的副作用。5、与正常组相比,模型组大鼠下丘脑CRH含量明显升高,两组间比较P<0.05,差异具有统计学意义。6、与模型组相比,西药组、中药高、中、低剂量组大鼠下丘脑CRH水平均有所降低。其中,西药组、中药康泰复方高、中剂量组与模型组相比P<0.05,差异均有统计学意义；中药低剂量组与模型组相比虽无统计学差异,但仍存在下降的趋势。7、与正常组相比,模型组大鼠脑干CRH含量明显升高,两组间比较P<0.05,差异具有统计学意义。8、与模型组相比,西药组、中药高、中、低剂量组大鼠脑干CRH水平均有所降低。其中,西药组、中药康泰复方高、中剂量组与模型组相比P<0.05,差异均有统计学意义；中药低剂量组与模型组相比虽无统计学差异,但仍存在下降的趋势。9、光镜下观察c-Fos表达染色呈棕黄色,与正常组相比,模型组大鼠结肠远端肥大细胞c-fos表达水平明显升高,两组间比较P<0.05,差异具有统计学意义。10、与模型组相比,西药组、中药高、中、低剂量组大鼠结肠远端肥大细胞c-fos基因表达水平均有所降低。其中,西药组、中药康泰复方高、中、低剂量组与模型组相比P<0.05,差异均有统计学意义；并且,中药中药康泰复方用药剂量与鼠结肠远端肥大细胞c-fos基因表达水平呈负相关。结论1.D-IBS大鼠脑干、下丘脑CRH含量表达的升高提示其表达改变与D-IBS的发病相关。2.中药康泰复方是治疗IBS-D安全有效的方剂。3.基于心胃相关理论组方的中药康泰复方能降低IBS-D高敏性大鼠脑干、下丘脑CRH含量,其作用机制可能是与影响脑肠轴CRH通路有关。4、用科学的研究方法的出实验结果证实了周福生教授提出的“心胃相关理论”的正确性。更多还原

【Abstract】 Irritable bowel syndrome is a common functional gastroenteropathy which is the characteristics of abdominal pain/abdominal discomfort and bowel evacuation habit, is short of change of morphology and biochemics. Its pathogenesis isnot clear so far. Now generally agreed that a lot of factors play an important role in clausing smooth muscle dyskinesia and visceral paresthesia, specifically mentality, psyche, society and circumstances, ect. This article reviews the onset, pathopoiesis, diagnosis and treatment of IBS, and relatted to gut hormones and Corticotropin-releasing hormone (CRH), MC to IBS.OBJECTIVEThrough our animal experiments, we aim to prove that the mechanism of irritable bowel syndrome (IBS) is related with the expression of mRNA on CRH and CRH-R1in colonic mucosas. As well as deepening our understanding of the pathogenesis mechanisms of irritable bowel syndrome, it can help us to explain the pathway and mechanism of the treatment of irritable bowel syndrome with Chinese medicine Kangtai compound, and provide the scientific basis and theoretical support for the theory of the stomach treatment of irritable bowel syndrome in the modernbiology.METHODSA total of110Sprague-Dawley rats7D were randomly divided into normal control group and experimental group, whereas100in the experimental group. A rat model of IBS was established by Separation of mother and child plus rectal stimulation in young rats. Then the SD rats in the experimental group were randomly divided into model group、KangTai low dose group of TCM、KangTai dose group of TCM、KangTai high dose group of TCM and Trimebutine Maleate group by weight. After a treatment for two weeks, remove the thalamus, brain stem, testing CRH level and MC in Appendix.RESULTS:1in normal rats and model rats at different times body weightSeparation of mother and child with young rats increases visceral sensitivity to rectal stimulation in IBS rat reproduction and normal control rats no significant changes in body weight differences (P>0.05), shows the modeling of this Act does not affect the normal rats.2rats in the awake state, the gradual expansion of intestinal pressure, the abdomen of rats raised the blood pressure with the back arched pressure, on average, increase the use of separation of mother and pups copy rectal stimulation in IBS visceral hypersensitivity in rats model, in its sixth week after birth in rat visceral hypersensitivity was significantly higher (P <0.05), shows that successful model.3, IBS visceral hypersensitivity rats Kangtai capsule after2weeks of treatment, medication visceral hypersensitivity in rats significantly reduced (P<0.05), and middle and high dose treatment group had returned to near normal levels (P>0.05.) Tip Kangtai capsule on rat model of IBS improve the role of visceral hypersensitivity.4, Kangtai capsule treatment, separation of mother and child with young rats increases visceral sensitivity to rectal stimulation in IBS rat reproduction and normal control group, body weight, activity, shiny hair no significant difference (P>0.05), shows Kangtai capsule is safety, there is no obvious side effects.5, compared with the normal group, model group were significantly increased hypothalamic CRH between the two groups P<0.05, statistically significant difference.6, compared with model group, medication group, Chinese high, medium and low-dose group hypothalamic CRH levels were decreased. Among them, the western medicine group, traditional Chinese medicine compound Conde high dose group compared with model group, P<0.05, differences were statistically significant; medicine low dose group compared with model group although no significant difference, but there are still a downward trend 7, compared with the normal group, model group were significantly higher CRH brain stem between the two groups P<0.05, statistically significant difference.8, compared with model group, medication group, Chinese high, medium and low-dose group brainstem CRH levels were decreased. Among them, the western medicine group, traditional Chinese medicine compound Conde high dose group compared with model group, P<0.05, differences were statistically significant; medicine low dose group compared with model group although no significant difference, but there are still a downward trend9, light microscopy c-Fos expression showed brown staining, compared with normal rats, the distal colon mast cell c-fos expression levels were significantly increased between the two groups P<0.05, statistical difference significance.10, compared with model group, medication group, Chinese high, medium and low-dose group distal colon mast cells c-fos gene expression levels were decreased. Among them, the western medicine group, traditional Chinese medicine compound Conde high, medium and low-dose group compared with model group P<0.05, differences were statistically significant; and traditional Chinese medicine traditional Chinese medicine compound dosage Conde distal colon mast cells with mouse c-fos gene expression levels were negatively correlated.CONCLUSION:1. D-IBS rat brain stem, hypothalamus CRH expression levels lower threshold and prompted changes in the expression of the pathogenesis of D-IBS.(2) traditional Chinese medicine treatment of Conde IBS-D compound is safe and effective prescription.3groups based on the theory of the heart side of the stomach medicine Conde compound can reduce IBS-D high-sensitivity in rat brain stem, hypothalamus CRH content, and its mechanism may affect the brain-gut axis with CRH pathway.4, using scientific research methods of the experimental results confirm Professor Zhou Fusheng’s "heart-related stomach theory" is correct.更多还原