当归对血虚便秘模型小鼠结肠水通道蛋白8表达的影响

摘要

Objective To observe the effects of Angelica Sinensis Radix on the expression of aquaporin 8 (AQP8) and the levels of AC-cAMP-PKA in colon of blood-deficiency constipation in mice; To identify mechanism of Angelica Sinensis Radix for loosening the bowels to relieve constipation. Methods Sixty Kunming mice were randomly divided into control group, model group, positive medicine group, and Angelica Sinensis Radix high-, medium-, and low-dose groups, with ten mice in each group. Diphenoxylate, acetylphenlyhydrazine and cyclophosphamide were used to establish blood-deficiency constipation mice models. From the 14thday of the experiment, Angelica Sinensis Radix high-, medium-, and low-dose groups were given 16.7, 8.8, and 4.2 g/kg Angelica Sinensis Radix Decoction for gavage. Positive medicine group was given 5.0 g/kg Changtongshu Granules Liquid for gavage. Control group and model group were given equal volume of saline for gavage. The symptoms of blood deficiency constipation were observed and defecation time. Immunohistochemistry, Western blot and qRT-PCR were used to detect the expression of AQP8 protein and mRNA in colon, and the expression of AC-cAMP-PKA in colon was detected by ELISA. Results The mice in the model group developed blood deficiency constipation syndrome; the defecation time was significantly prolonged (P＜0.01); the expression level of colonic AQP8 protein and mRNA, AC, cAMP and PKA significantly increased (P＜0.05, P＜0.01). Compared with model group, the defecation time was significantly shortened in Angelica Sinensis Radix high-, medium-, and low-dose groups; the expression of AQP8 protein and mRNA and the levels of AC, cAMP and PKA were significantly decreased (P＜0.05, P＜0.01). Conclusion The treatment of Angelica Sinensis Radix for blood-deficiency constipation may be related to adjusting the AC-cAMP-PKA signaling pathways and reducing the expression of AQP8 protein and mRNA.%目的 观察当归对血虚便秘模型小鼠结肠水通道蛋白8(AQP8)的表达及AC-cAMP-PKA信号通路各因子含量的影响,探讨当归润肠通便作用机制.方法 60只KM小鼠随机分为正常组、模型组、阳性药组和当归高、中、低剂量组,每组10只.采用复方地芬诺酯、乙酰苯肼及环磷酰胺联合复制血虚便秘小鼠模型,实验第14日起,当归高、中、低剂量组分别给予16.7、8.8、4.2 g原药材/kg当归水煎液灌胃,阳性药组给予5.0 g/kg常通舒颗粒药液灌胃,正常组和模型组给予等体积生理盐水灌胃,观察小鼠血虚便秘证候、排便时间,免疫组化、Western blot和qRT-PCR检测小鼠结肠AQP8蛋白和mRNA的表达,ELISA检测小鼠结肠AC-cAMP-PKA信号通路中各因子的表达.结果 模型组小鼠出现血虚便秘证候,粪便排出时间较正常组明显延长(P＜0.01),结肠AQP8蛋白和mRNA表达及腺苷酸环化酶(AC)、环磷酸腺苷(cAMP)、蛋白激酶A (PKA)含量显著升高(P＜0.05,P＜0.01);与模型组比较,当归各剂量组小鼠粪便排出时间明显缩短,结肠AQP8蛋白和mRNA表达及AC、cAMP、PKA含量显著降低(P＜0.05,P＜0.01).结论 当归治疗血虚便秘可能与调节结肠AC-cAMP-PKA信号通路,下调结肠AQP8蛋白和mRNA的表达有关.