基于大肠主津理论下助阳通便膏对便秘模型小鼠结肠组织中MUC2、AQP3的影响

Effect of Zhuyang Tongbian Cream on MUC2 and AQP3 in Colon Tissue of Constipation Mice Based on Theory of Large Intestinal Dominating Body Fluid

目的通过观察助阳通便膏对便秘模型小鼠结肠黏膜蛋白2(MUC2)、水通道蛋白3(AQP3)的影响,探讨助阳通便膏改善功能性便秘的作用机制。方法将120只雄性巴比西小鼠随机分成5组,每组24只。即正常组、模型组、助阳通便汤组、助阳通便膏组、麻仁软胶囊组。除正常组外,其余各组造模便秘小鼠模型,采用复方地芬诺酯灌胃的方法造模。造模成功后各组分别给予相应的药物,连续灌胃给药14 d后取材,应用免疫组化、Wester Blot的方法观察各组小鼠结肠黏膜蛋白2(MUC2)和水通道蛋白3(AQP3)的含量。结果通过免疫组化的方法发现,与模型组比较,各组MUC2、AQP3平均光密度值均有不同程度增高,且助阳通便膏组与汤剂组比较MUC2平均光密度值差异有统计学意义(P<0.05)。通过Wester Blot的方法发现,与模型组比较,各组MUC2、AQP3蛋白相对表达水平灰度值均有不同程度增高。且AQP3助阳通便膏剂组与汤剂组比较(P<0.05)。结论助阳通便膏可提高MUC2、AQP3含量,改善便秘。助阳通便膏剂增加便秘小鼠结肠AQP3蛋白表达水平的作用及增加便秘小鼠结肠MUC2阳性细胞数量的作用明显优于助阳通便汤。

Objective To investigate the effects of Zhuyang Tongbian Cream on colonic mucoprotein 2(MUC2)and Aquapor3(AQP3)in constipation mice and investigate the mechanism of relieving functional constipation.Method A total of 120 male Babesi mice were randomly divided into 5 groups:the normal group,the model group,the Zhuyang Tongbian Decoction group,the Zhuyang Tongbian Cream group and the Maren Soft Capsule group.In addition to the normal group,the model of constipation in other groups was set up by the method of compound difenoester lavage.After the successful modeling,each group was given the corresponding drugs for 14 days.The contents of MUC2 and AQP3 in mice were observed by immunohistochemical and Western blot methods.Results By immunohistochemical method,it was found that the average light density values of MUC2 and AQP3 in each group were increased to varying degrees compared with those of the model group,and the MUC2 average light density values of the Zhuyang Tongbian Cream group was better than that of the Zhuyang Tongbian Decoction group(P<0.05).Through the method of Western blot,it was found that compared with those of the model group,the relative expression levels of MUC2 and AQP3 proteins in each group increased to varying degrees.AQP3 of the Zhuyang Tongbian Cream group was better than that of the Zhuyang Tongbian Decoction group(P<0.05).Conclusion Zhuyang Tongbian Cream can increase the contents of MUC2 and AQP3 and relieve constipation.The effect of Zhuyang Tongbian cream on the expression level of AQP3 protein and the number of MUC2 positive cells in colon of constipation mice was better than that of Zhuyang Tongbian Decoction.