基于津液理论探讨改良枳术方对便秘大鼠AQP3/AQP9、MUC2的影响＊

目的 在津液理论指导下探讨改良枳术方对于便秘大鼠AQP3/AQP9、MUC2的影响。方法 48只Wistar大鼠随机分为空白组、模型组、莫沙必利组、改良枳术方低、中、高剂量组,每组8只。除空白组外,其他各组均采用洛哌丁胺灌胃制备慢传输便秘大鼠模型。改良枳术方低、中、高剂量组大鼠分别给予4.725、7.875、11.024 g/（kg·d）灌胃,莫沙必利组以1.56 mg/（kg·d）莫沙必利混悬液灌胃,空白组和模型组灌胃等剂量的蒸馏水。每天一次,共持续7天。检测各小组大鼠24 h粪便颗粒数,粪便含水量、肠道转运时间、肠道推进率、免疫组化和蛋白免疫印迹法（Western blot, WB）测定结肠水通道蛋白3（Aquaporin3, AQP3）、水通道蛋白9（Aquaporin9, AQP9）、黏蛋白2（Mucin 2, MUC2）蛋白表达。结果 与空白组比,模型组大鼠24 h粪便颗粒数及粪便含水量明显减少、肠道转运时间延长、肠道推进率降低,结肠AQP3蛋白表达升高、MUC2蛋白表达降低（P<0.05）;与模型组比,改良枳术方各组大鼠24 h粪便颗粒数及粪便含水量明显增加、肠道推进率增加,改良枳术方各组结肠AQP3表达下降、MUC2蛋白表达增加（P<0.05）;与模型组比,改良枳术方高剂量组结肠AQP9蛋白表达明显下降（P<0.05）。结论 AQP3/AQP9、MUC2介导的结肠“津”“液”代谢在便秘发生发展中发挥重要作用,改良枳术方用于便秘疗效确切,其中以中、高剂量效果更佳,可能的作用机制为通过AQP3/AQP9、MUC2从而调控结肠的“津”“液”代谢。

Effect of Modified Zhizhu Decoction on AQP3/AQP9 and MUC2 in Rats with Constipation Based on Body Fluid Theory

Objective To explore the effect of modified Zhizhu Decoction on AQP3/AQP9 and MUC2 in constipation rats under the guidance of body fluid theory.Methods 48 Wistar rats were randomly divided into blank group, model group, mosapride group, modified Zhizhu Decoction low, medium and high dose group, with 8 rats in each group. Except for the blank group, the other groups were given loperamide to prepare slow transit constipation rat model. The rats in low, middle and high dose groups of modified Zhizhu Decoction were given 4.725, 7.875 and 11.024 g/(kg·D) by gavage respectively. The rats in mosapride group were given 1.56 mg/(kg·D) mosapride suspension by gavage. The rats in blank group and model group were given the same dose of distilled water by gavage. Once a day for 7 days. The number of fecal particles, fecal water content, gastrointestinal transit time, intestinal propulsion rate and the expression of AQP3, AQP9 and MUC2 protein in colon were detected by immunohistochemistry and western blot.Results Compared with the blank group, the number of fecal particles and fecal water content were significantly reduced, the intestinal propulsion rate was decreased, the expression of AQP3 protein was increased, and the expression of MUC2 protein was decreased in the model group (P<0.05). Compared with the model group, the expression of AQP3 decreased and the expression of MUC2 increased in each dose group of modified Zhizhu Decoction (P<0.05),the expression of AQP9 protein in colon of high-dose modified Zhizhu Decoction group was significantly decreased (P<0.05).Conclusion AQP3/AQP9, MUC2 mediated colonic "jin and ye" metabolism plays an important role in the occurrence and development of constipation. The modified Zhizhu Decoction has a definite effect on constipation, and the effect of middle and high dose is better. The possible mechanism is to affect colonic "jin and ye" metabolism by regulating AQP3 / AQP9, MUC2.