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目的:探讨养阴润肠方对便秘小鼠结肠水通道蛋白3(AQP3)和水通道蛋白9(AQP9)表达的影响。方法:50只雄性ICR小鼠随机分为正常组,模型组,养阴润肠方低、高剂量组,普卢卡必利组,每组10只,采用复方地芬诺酯20 mg·kg~(-1)连续灌胃15 d建立便秘模型,每周计算粪便含水量,用养阴润肠方低、高剂量组(29.6,59.2 g·kg~(-1))干预15 d,取材结肠组织,进行免疫组化AQP3和AQP9定位和半定量表达,实时荧光定量聚合酶链式反应(Real-time PCR),蛋白免疫印迹法(Western blot)检测AQP3和AQP9基因和蛋白表达情况。结果:与正常组比较,模型组粪便含水量明显下降(P<0.01),AQP3表达增加(P<0.01),AQP3的mRNA和蛋白表达均增加(P<0.05),AQP9表达减少,AQP9的mRWA和蛋白表达亦均下降,但无统计学差异;与模型组比较,养阴润肠方低、高剂量组粪便含水量均增加(P<0.05),AQP3表达下降(P<0.05),AQP3的mRNA和蛋白表达均下降(P<0.05);养阴润肠方低剂量组的AQP9表达增加(P<0.01),AQP9的mRNA和蛋白表达均升高(P<0.05),高剂量组表达有升高趋势,但无统计学差异。结论:养阴润肠方可以增加便秘小鼠粪便含水量,其机制可能与降低结肠组织中AQP3和增加AQP9的表达有关,并且大剂量作用反而不明显。
Objective: To investigate the effect of Yangyin Runchang Formula on colon aquaporin 3 (AQP3) and aquaporin 9 (AQP9) expression in constipation mice. Methods: 50 male ICR mice were randomly divided into normal group, model group, yin yang runchang low, high dose group and prucalopride group, 10 rats in each group. The compound diphenoxylate 20 mg · kg ~ (-1) for 15 days, the constipation model was established and the water content of feces was calculated weekly. After intervention for 15 days with low-dose and high-dose Yangyin Runchang group (29.6, 59.2 g · kg -1) Colon tissues were harvested for immunohistochemical localization and semi-quantitative expression of AQP3 and AQP9. Real-time PCR and Western blot were used to detect the expression of AQP3 and AQP9 mRNA and protein. Results: Compared with the normal group, the water content of feces in model group decreased significantly (P <0.01), the expression of AQP3 increased (P <0.01), the mRNA and protein expression of AQP3 increased, the expression of AQP9 decreased (P <0.05), the expression of AQP3 was decreased (P <0.05), the expression of AQP3 was lower in model group than in model group (P <0.05). The expression of AQP9 in low-dose of Yin-Yin Runchang prescription group was increased (P <0.01), and the expression of AQP9 mRNA and protein were higher in high-dose group Increasing trend, but no statistical difference. Conclusion: Yangyin Runchang can increase water content in feces of mice with constipation. The mechanism may be related to the decrease of AQP3 and the expression of AQP9 in colonic tissues, but the effect of high dose is not obvious.