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目的 探讨一氧化氮 (NO)在肠易激综合征 (IBS)发病机制中的作用 ,并从基因水平揭示NO含量改变的原因。方法 (1)应用电子气压泵及灌注导管测压仪研究 2 5例腹泻型IBS患者及 15例正常志愿者的肛门、直肠压力、直肠顺应性、乙状结肠和直肠运动指数以及直肠对容量刺激的感觉阈值 ;(2 )应用硝酸还原酶法测定两组肠黏膜NO的含量 ;(3)NADPH黄递酶组化法和计算机图像分析系统对两组肠黏膜肌层一氧化氮合酶 (NOS)阳性神经纤维作定量分析 ;(4)采用荧光定量PCR(FQ PCR)方法对神经型一氧化氮合酶 (nNOS)的基因表达进行定量分析。结果 (1)肠道测压 :IBS患者的直肠静息压、肛管上部静息压、收缩压、松弛压、肛管下部静息压、收缩压、松弛压和直肠顺应性与正常人比较 ,差异无显著性 (P >0 .0 5 ) ;患者乙状结肠和直肠运动指数明显高于正常人 (P <0 .0 5 ) ;(2 )直肠内脏感觉阈值 :最低感觉阈值、排便阈值和疼痛阈值明显低于正常人 (P <0 .0 5 ) ;(3)肠黏膜NO含量 :患者结肠黏膜NO含量显著低于正常人 ,并且患者的NO含量与运动指数成负相关 ,与感觉阈值、排便阈值、疼痛阈值呈正相关 (P <0 .0 5 ) ;(4)NADPH组化染色 :IBS患者黏膜肌层NOS阳性神经纤维的面积和平均吸光度较正常人显著减少 (P <0 .0 5 ) ;(5 )NOS mRNA
Objective To investigate the role of nitric oxide (NO) in the pathogenesis of irritable bowel syndrome (IBS) and to reveal the cause of the change of NO content at the gene level. Methods (1) The effects of anus, rectal pressure, rectal compliance, sigmoid and rectal movement index, and sensation of rectum volume stimulation on 25 patients with diarrhea-type IBS and 15 normal volunteers were studied by electronic pressure pump and perfusion catheter manometer. (2) Nitric acid reductase method was used to determine the content of NO in the two groups of intestinal mucosa; (3) NADPH diaphorase histochemistry and computer image analysis system were used to detect the positive expression of nitric oxide synthase (NOS) (4) Quantitative analysis of the gene expression of neuronal nitric oxide synthase (nNOS) by fluorescence quantitative PCR (FQ PCR). Results (1) Intestinal pressure: IBS resting pressure, anal upper resting pressure, systolic pressure, relaxation pressure, anal lower resting pressure, systolic pressure, relaxation pressure and rectal compliance compared with normal people , The difference was not significant (P> 0.05); The sigmoid colon and rectal movement index were significantly higher in patients than in normal people (P <0.05); (2) The sensory threshold of rectum viscera: the lowest sensory threshold, defecation threshold and pain (3) The content of NO in mucosa: the content of NO in colonic mucosa of patients was significantly lower than that of normal subjects, and the content of NO in patients was negatively correlated with the index of exercise, and the threshold of sensory threshold, Defecation threshold and pain threshold were positively correlated (P <0.05). (4) NADPH staining: The area and average absorbance of NOS positive nerve fibers in mucosa of IBS patients were significantly lower than those in normal controls (P <0.05) ); (5) NOS mRNA