【作者】 陈吉；

【导师】 邓展生；

【作者基本信息】 中南大学 ， 外科学， 2012， 博士

【摘要】 第一部分肠易激综合征患者肠粘膜脑肠肽及肥大细胞的变化目的：探讨IBS患者肠粘膜血管活性肠肽(VIP)、降钙素基因相关肽(CGRP)和肥大细胞(MC)的变化,以及它们在IBS中的作用和临床意义。方法：参照罗马Ⅲ诊断标准,选择腹泻型IBS (D-IBS)、便秘型IBS (C-IBS)和健康对照组各20例,每例各取回盲部、乙状结肠粘膜4块,应用免疫组织化学染色法分别检测VIP和CGRP; MC检测采用甲苯胺蓝染色法,去观察它们的阳性神经纤维表达。结果：D-IBS、C-IBS和患者VIP、CGRP免疫阳性神经纤维及MC阳性神经纤维表达量明显高于正常对照组(P<0.01),且回盲部与乙状结肠比较无显著性差异(P>0.05),而C-IBS患者回盲部CGRP、 MC与乙状结肠比较无显著性差异。结论：VIP、CGRP和MC可能参与了IBS的发病的病理生理过程。第二部分肠易激综合征患者肠粘膜细胞因子Thl/Th2的漂移目的：探讨IBS患者肠粘膜中Th1细胞因子干扰素(IFN)-γ、白细胞介素(IL)-12与Th2细胞因子IL-4、IL-10的表达情况,以阐述IBS患者肠粘膜中是否存在Th1/Th2免疫调节功能紊乱及其可能机制。方法：按照罗马Ⅲ标准,选择60例IBS患者,其中腹泻型IBS (D-IBS)38例,其中感染后IBS (PI-IBS)20例,非感染后IBS (non-PI-IBS)18例；便秘型IBS (C-IBS)22例；健康对照20例。经结肠镜检查钳60例患者和健康对照组的升结肠、降结肠及直肠粘膜标本。采用RT-PCR和Western blot方法检测肠道各部位肠粘膜中Th1、Th2细胞因子mRNA与蛋白的表达水平。结果：(1)D-IBS患者肠粘膜中Th1细胞因子IFN-γ、IL-12及Th2细胞因子IL-4mRNA与蛋白的表达水平较对照组差异无显著性(P>0.05),而Th2细胞因子IL-10mRNA与蛋白的表达水平显著低于对照组(P<0.05), C-IBS与健康对照比较差异无显著性(P>0.05),C-IBS与健康对照比较差异均无显著性。(2)PI-IBS患者肠粘膜中Thl细胞因子IFN-γ mRNA与蛋白的表达水平较对照组比较显著增高,差异有显著性(P<0.05)；Th2细胞因子IL-10mRNA与蛋白的表达量显著降低,差异有显著性(P<0.05)；而Thl细胞因子IL-12与Th2细胞因子IL-4mRNA与蛋白的表达水平与对照组比较,差异无显著性(P>0.05), non-PI-IBS与健康对照比较差异均无显著性。(3)各组不同部位肠粘膜内Th1、Th2细胞因子表达水平比较,差异均无显著性。结论：(1)D-IBS患者肠粘膜中Th2抗炎细胞因子IL-10表达水平降低,存在Th1/Th2漂移,Th2反应呈弱势,相对而言Thl免疫反应占优势。(2)C-IBS患者Thl/Th2处于免疫平衡状态。(3)PI-IBS患者肠粘膜内Thl细胞因子IFN-γ表达增强,Th2细胞因子IL-10表达减弱,存在Thl/Th2漂移,明显表现为Thl免疫反应呈优势,Th2反应呈弱势。更多还原

【Abstract】 Part I Changes of brain-gut peptide and mast cell in irritable bowel-syndromeObjective:To investigate the changes of vasoactive intestinal polypeptide (VIP)、calcitonin gene-related peptide (CGRP) and mast cell (MC) in irritable bowel syndrome (IBS), and to study their possible roles in pathogensis.Methods:In40patients with IBS who fulfilled the Rome Ⅲ criteria and in20non-IBS control subject, biopsy specimens taken from the mucosa of terminal ileum and recto-sigmoid junction at colouscopy were further examined using immunohistochemical staining of VIP, CGRP and MC. The results of immunohistochemisty were investigated qualitatively by color image analyzer.Results:Significant increase of VIP, CGRP and MC were found in IBS of colon compared to the controls (P<0.01), and compared with sigmoid, there were no significantly difference in ileum. But the iluem and sigmoid compared, CGRP and MC were no significantly difference in C-IBS patients (P>0.05).Conclusion:VIP、CGRP and MC are involved in the pathophysiological mechanism of IBS in certain ways, and their possible roles in the colon suggests the further studies may be useful in the understanding of IBS pathophysiology. Part Ⅱ Study on the shifting of Thl/Th2balance of large intestinal mucosa in patients with irritable bowel syndromeObjective:To investigate the expressions of Thl cytokine IFN-γ、 interleukin(IL)-12and Th2cytokine IL-4、IL-10intestinal mucosa in patients with irritable bowel syndrome(IBS). The primary of this study was to demonstrate immune mechanism underlying the activation of Thl/Th2cytokine production in colonic mucosa, so as to provide new idea for IBS in humans.Methods:The patients with IBS were diagnosed according to Rome Ⅲ criteria. Mucosal speciments were harvested from ascending, descending colon and rectum by colonscopy in60patients with IBS and20health controls. The expressions of Thl and Th2cytokine gene expressions and protein secretion were examined by RT-PCR and Western blot respectively.Results:(1) Compared with controls, the gene expressions and protein secretion of Thl cytokine IFN-γ、IL-12and IL-4have no significant difference. However, the expressions of Th2cytokine IL-10in diarrhea-predominant IBS (D-IBS) were decreased significantly compared with controls. In constipation predominant IBS (C-IBS) patients, the expressions of all Th1and Th2cytokine had no statistical difference compared with control.(2) The gene expression and protein secretion of IFN-γ in post infective IBS (PI-IBS) patients were increased significantly increased. The expressions of IL-4and IL-12in PI-IBS no significant difference compared with controls. However, the expressions of IL-10in PI-IBS patients were decreased significantly compared with control. The expressions of Thl and Th2cytokine in non-PI-IBS patients had no statistical difference compared with controls.(3) The expressions of Thl cytokine IFN-γ、IL-12and Th2cytokine IL-4、IL-10had no statistical difference compared with each other in all parts of gut.Conclusion:(1) Because the expressions of Th2IL-10mRNA and protein were decreased significantly, the shift of Thl/Th2balance of colon and rectal mucosa in D-IBS patients was demonstrated along with decreased Th2activity.(2) There are Th1/Th2immune balance in C-IBS patients.(3) Because increased expression of proinflammatory cytokine Thl IFN-γ and decreased expression of anti-inflammatory cytokine Th2IL-10mRNA and protein, the shift of Thl/Th2balance of colon and rectal mucosa in PI-IBS patients was demonstrated along with increased Th1and decreased Th2immune activity.更多还原