【作者】 陈丽珠；

【导师】 王承党；

【作者基本信息】 福建医科大学 ， 内科学， 2011， 硕士

【摘要】 目的:检测腹泻型肠易激综合征(IBS-D)患者和健康对照者升结肠黏膜类胰蛋白酶、α1-抗胰蛋白酶的表达水平,并探讨它们与IBS-D症状的关系。方法:符合RomeⅢ标准的IBS-D患者20例、健康对照组10例纳入研究,在接受电子肠镜检查时取升结肠黏膜标本3块,称质量、生理盐水漂洗后迅速在2ml HBSS平衡盐液中37℃下孵育1 h,ELISA法和Western blotting法检测孵育上清液中类胰蛋白酶、α1-抗胰蛋白酶水平, RT-PCR法检测升结肠黏膜类胰蛋白酶、α1-抗胰蛋白酶mRAN表达。类胰蛋白酶、α1-抗胰蛋白酶表达水平与IBS-D患者的胃肠道症状评分进行相关性分析。结果:1、ELISA法结果:IBS-D患者升结肠黏膜组织孵育上清液中类胰蛋白酶浓度为(0.070±0.011)ug/g组织,健康对照组为(0.043±0.006) ug/g组织,差异有统计学意义(P=0.000);而α1-抗胰蛋白酶浓度为(2.36±1.05mg/g组织)与健康对照组为(2.68±1.19 mg/g组织)的差异无统计学意义(P=0.455)。类胰蛋白酶浓度与胃肠道症状评分(GSRS)呈正相关( r =0.592, P =0.008)。2、RT-PCR法结果:IBS-D患者升结肠黏膜组织类胰蛋白酶相对表达度为0.83±0.06,健康对照组为0.58±0.13,差异有统计学意义( P =0.000);而α1-抗胰蛋白酶相对表达度为(1.02±0.19)与健康对照组为(0.97±0.17)的差异无统计学意义( P =0.413)。RT-PCR法检测的类胰蛋白酶相对表达度与GSRS也呈正相关(r=0.520, P =0.009)。3、Western blotting法结果:IBS-D患者升结肠黏膜组织类胰蛋白酶相对浓度为0.53±0.09,健康对照组为0.23±0.08,差异有统计学意义(P=0.000);而α1-抗胰蛋白酶相对浓度为(0.81±0.15)与健康对照组为(0.80±0.11)的差异无统计学意义(P=0.761)。Western blotting法测定的类胰蛋白酶相对浓度与GSRS呈正相关( r =0.421,P=0.032)。结论:腹泻型肠易激综合征患者升结肠黏膜中类胰蛋白酶的表达水平增加,而α1-抗胰蛋白酶表达无明显差异,这种变化与IBS-D的消化道症状相关,因此,升结肠肥大细胞中类胰蛋白酶表达增加可能与IBS-D症状的发生机制有关。更多还原

【Abstract】 Aim: To detect the contents of tryptase andα1-antitrypsin in the ascending colonic mucosa in patients with diarrhea-predominant irritable bowel syndrome (IBS-D), and to explore their role in the pathogenesis of this disorder.Methods: Twenty patients with IBS-D fulfilling the Rome III criteria and ten healthy volunteers were included in this study. Three biopsy samples were collected from the ascending colon during the colonoscopy, and then incubated in 2ml HBSS 37℃for 1h. The concentrations of tryptase andα1-antitrypsin in the supernatants were detected by enzyme-linked immunosorbent assay (ELISA). The proteins and mRNA of tryptase andα1-antitrypsin in the mucosa were detected by Western blotting and RT-PCR respectively.The relationship between gastrointestinal symptom rating scale (GSRS) and the contents of tryptase in the ascending colon were also analyzed.Results: The contents of tryptase in ascending colon mucosa supernatants of the IBS-D patients detected by ELISA were (0.070±0.011)ug/g tissue,while in the healthy volunteers were (0.043±0.006) ug/g tissue. There was a significant difference between them( P =0.000). Hower, the mean content ofα1-antitrypsin in the supernatants of the IBS-D patients (2.36±1.05)mg/g tissue detected by ELISA was not different from that in the healthy volunteers (2.68±1.19)mg/g tissue ( P=0.455). The contents of tryptase were positively correlated with GSRS score( r = 0.592, P = 0.008).The relative expression of tryptase mRNA in the ascending colon mucosa of the IBS-D patients detected by RT-PCR was 0.83±0.06,while in the healthy volunteers was 0.23±0.08. The difference has a statistical significance( P =0.000). The relative expression ofα1-antitrypsin mRNA in the ascending colon mucosa of the IBS-D patients detected by RT-PCR was 1.02±0.19,while in the healthy volunteers was 0.97±0.17. There was not a significant difference between them( P =0.413). The relative expression of mRNA of tryptase was positively correlated with GSRS score also( r = 0.520, P = 0.009).The relative expression of tryptase in the ascending colon mucosa of the IBS-D patients detected by Western blotting was 0.53±0.09,while in the healthy volunteers was 0.23±0.08. The difference of the results has statistical sense( P =0.000). The relative expression ofα1-antitrypsin in ascending colon mucosa of the IBS-D patients detected by Western blotting was 0.81±0.15,while in the healthy volunteers was 0.80±0.11. The difference of the results has no statistical sense( P =0.761). The relative expression of tryptase was positively correlated with GSRS score( r = 0.421, P = 0.032).Conclusion: The expression of tryptase in the ascending colon of IBS-D patients increased, but ofα1-antitrypsin not, and this change is related to gastrointestinal symptoms. Therefore, excessive expression of tryptase in the colon may be involved in the pathogenesis of IBS-D.更多还原