目的:分别建立大鼠血虚、阴虚和阳虚便秘模型并比较其结肠动力、结肠水代谢、结肠黏液分泌和结肠水通道蛋白2(AQP2)的表达水平。方法:40只SD大鼠,雌雄各半,随机分为4组(n=10):正常对照组(N)、血虚便秘组(xc)、阴虚便秘组(xc)、阳虚便秘组(YAC)。分别采用放血+洛哌丁胺、甲状腺素+洛哌丁胺、冰水刺激+洛哌丁胺建立血虚便秘、阴虚便秘、阳虚便秘大鼠模型,放血每周1次,药物每天灌胃1次,连续42d。检测大鼠活动状态、体质量、大便性状、口肛传输时间、小肠推进率、粪便和结肠含水量等体征指标,对结肠组织进行AB-PAS染色和免疫组化染色,观察黏液分泌情况和AQP2表达水平。结果:与正常对照组比较,3种模型大鼠体重增长减慢;造模第40天自主活动量变化率由大到小依次为YC、XC和YAC;造模第30天出现固体硬质粪便,粪便评分升高顺序为XC、YC和YAC;口肛传输时间均显著延长、小肠推进率均显著降低(P〈0.05,P〈0.01),肠动力减低顺序为YC、YAC和XC;粪便含水量均显著减少(P〈0.05,P〈0.01),XC和YAC组大鼠结肠含水量显著减少(P〈0.05,P〈0.01),YC大鼠结肠含水量有减少趋势,结肠水代谢顺序为YC、YAC和XC;3种模型大鼠结肠黏膜层腺管和杯状细胞出现不同程度缩小,黏液分泌减少,分泌功能障碍顺序为YAC、YC和XC,且近端和远端结肠AQP2的表达均显著增加(P〈0.05,P〈0.01),增加顺序近端结肠为YAc、YC、XC,远端结肠为YAC、XC和YC。结论:复合因素长期刺激均可建立符合中医体征的便秘大鼠模型,且各模型大鼠结肠运动、结肠水代谢、结肠黏液分泌和AQP2表达水平存在一定的差异。
Objective: To establish three types constipation models with the syndrome of Xue-Xu, Yin-Xu and Yang-Xu in rats, and to compare the difference of colonic motility, colonic water metabolism, colonic mucus secretion and the level of aquaporin-2 (AQP2). Methoils: Forty Sprague Dawley rats, male and female in half, were randomly divided into four groups: normal control group(N), Xue-Xu with constipation group(XC), Yin-Xu with constipation group(YC) and Yang-Xu with constipation group(YAC). Bloodletting and loperamide were used to induce the Xue-Xu with constipation in rats, thyroxin and loperamide were used to induce the Yin-Xu with constipation in rats, ice-water stimulation and loperamide were used to induce the Yang-Xu with constipation in rats. Bloodletting was eoducted once a week,drug was ad- ministrated orally to rats once a day for continuous 42 days. Rats' state, body weights, stool character, transfer time between mouth and anal, small intestinal propulsion rate were measured. The colon tissues of rats were stained with aleian blue-periodic acid schiff (AB-PAS) to analyze the changes of mucus secretion. The aquaporin-2 expressions in proximal and distal colon of rats were measured by immunohistochemical method. Results: Compared to normal control group, weight-increasing speed of the rats were reduced in three models. The order of independent movement change was YC, XC and YAC. Solid stool appeared at 30 d, fecal scores were increased. Transfer time between mouth and anal was significantly extended, small intestinal propulsion rate were significantly decreased( P 〈 0.05, P 〈 0.01 ). Water content in stool were significantly decreased in three type models( P 〈 0.05, P 〈 0.01), the content of water in colon were decreased in XC and YAC group (P 〈 0.05, P 〈 0.01). The colonic gland and goblet cells were narrowed in some extent, the mucus excretion were decreased. The levels of aquaperin-2 expressions in proximal and distal colon of rats were increased