枳术丸对脾虚证慢传输型便秘小鼠结肠PI3K、AKT表达的影响

Effects of Zhizhu Pills on expression of PI3K and AKT in colon of mice with slow transmission and constipation of spleen deficiency syndrome

目的:探讨枳术丸对脾虚证慢传输型便秘小鼠磷脂酰肌醇3-激酶(PI3K)、蛋白激酶B(AKT)表达的影响。方法:将47只SPF级小鼠随机分为正常组(15只)和造模组(32只)。造模组采用番泻叶灌胃、低纤维饮食、限食和限水的方法制备脾虚证慢传输型便秘模型,造模完成后,两组随机各取5只小鼠,进行模型验证。模型成功后,造模组随机分为模型组、枳术丸组和莫沙必利组,每组9只;正常组10只。枳术丸组按9.0g/kg灌胃,莫沙必利组按2.5mg/kg灌胃,正常组和模型组给予等容积的蒸馏水灌胃,4组灌胃容积均为0.1mL/10g,连续治疗7d。治疗完成后,采用免疫组化、Western Blot及Real-time PCR法检测各组PI3K、AKT蛋白及mRNA表达。结果:与正常组比较,模型组PI3K、AKT蛋白及mRNA表达均显著减少(P<0.01);与模型组比较,枳术丸组和莫沙必利组PI3K、AKT蛋白及mRNA表达均显著增加(P<0.01,P<0.05)。结论:脾虚证慢传输型便秘小鼠结肠组织中PI3K、AKT蛋白及基因表达均明显下降;枳术丸治疗脾虚证慢传输型便秘,可能与提高脾虚证慢传输型便秘小鼠结肠组织中PI3K、AKT表达水平有关。

Objective:To study the effect of Zhizhu Pills on the signal expression of phosphatidylinositol 3-kinase(PI3K)and protein kinase B(Akt)in mice with slow transit constipation of spleen deficiency.Methods:A total of 45 SPF mice were randomly divided into normal group(n=15)and model group(n=32).The slow transit constipation model of spleen deficiency syndrome was established by senna leaf,low fiber diet,food restriction and water restriction.After the model was established,5 mice in each group were randomly selected to verify the model.After the success of the model,the rats was randomly divided into model group(n=9),Zhizhu Pills group(n=9)and Mosapride group(n=9).10 rats in normal group Zhizhu Pills group was given 9.0 g/kg,Mosapride group was given 2.5 mg/kg,normal group and model group were given the same volume of distilled water,four groups were given 0.1 mL/10 g,continuous treatment for 7 days.After the treatment,the expression of PI3K,AKT protein and mRNA were detected by immunohistochemistry,Western Blot and Real-time PCR.Results:Compared with the normal group,the expression of PI3K,AKT protein and mRNA in the model group decreased significantly(P<0.01);Compared with the model group,the expression of PI3K,AKT protein and mRNA in the Zhizhu Pills group and Mosapride group increased significantly(P<0.01,P<0.05).Conclusion:The expression of PI3K,AKT protein and gene in the colon of mice with slow transit constipation of spleen deficiency syndrome were significantly decreased;Zhizhu Pills could improve the expression of PI3K and AKT in the colon of mice with slow transit constipation of spleen deficiency syndrome.