参苓白术颗粒对功能性腹泻大鼠结肠黏膜紧密连接蛋白表达的作用

Effect of Shenling Baizhu Particles on Intestinal Epithelial Light Junctions in Rats with Functional Diarrhea

目的:观察参苓白术颗粒对功能性腹泻模型大鼠肠黏膜紧密连接蛋白的影响,并探讨其作用机制。方法:采用高乳糖+小平台站立法建立功能性腹泻脾虚证大鼠模型,将90只大鼠随机分为正常组,模型组(高乳糖+小平板),匹维溴铵组,参苓白术颗粒高、中、低剂量组,每组15只,参苓白术颗粒高、中、低剂量分别为3.212,1.606,0.803 g·kg^(^(-1)),匹维溴铵给药剂量为0.012 5 g·kg^(^(-1));正常组与模型组灌服生理盐水,每天灌胃3次,共14 d。观察大鼠一般情况,采用酶联免疫吸附测定(ELISA)法检测血清中二胺氧化酶(DAO)水平、结肠黏膜肿瘤坏死因子-α(TNF-α)含量,实时荧光定量聚合酶链式反应(q PCR)法检测紧密连接蛋白^(^(-1))(zonula occludens^(^(-1)),ZO^(^(-1))),封闭蛋白^(^(-1))(Claudin^(^(-1))),咬合蛋白(Occludin)mRNA表达,免疫组化法检测ZO^(^(-1)),Occludin蛋白表达水平。结果:与正常组比较,模型组血清中DAO水平、结肠黏膜TNF-α含量明显升高(P<0.01),ZO^(^(-1)),Claudin^(^(-1)),Occludin mRNA表达明显降低(P<0.01),ZO^(^(-1)),Occludin蛋白表达明显降低(P<0.05);与模型组比较,参苓白术颗粒能够降低血清中DAO水平、结肠黏膜TNF-α含量(P<0.01),上调功能性腹泻模型大鼠结肠黏膜ZO^(^(-1)),Claudin^(^(-1)),Occludin mRNA表达(P<0.01),上调ZO^(^(-1)),Occludin蛋白表达(P<0.05,P<0.01)。结论:参苓白术颗粒能够调节功能性腹泻大鼠结肠黏膜紧密连接蛋白表达,调节肠黏膜机械屏障,这可能是从脾论治功能性腹泻的机制之一。

Objective： To discuss the effect of Shenling Baizhu particles on intestinal epithelial tight junctions in rats with functional diarrhea. Method： The functional diarrhea spleen deficiency syndrome model in rats was established by using high lactose ＋ small platform standing method. the rats were divided into normal group,model group,pinaverium bromide group,Shenling Baizhu particle high,medium and low dose groups.Shenling Baizhu particle groups were administrated with Shenling Baizhu particle at doses of 3. 212,1. 606,0. 803g·kg-1,respectively,and pinaverium bromide group was given pinaverium bromide of 0. 0125 g ·kg-1;meanwhile,normal group and model group were given physiological saline intragastrically for 14 successive days.The diamine oxidase（ DAO） and tumor necrosis factor-α（ TNF-α） were detected by enzyme-linked immunosorbent assay（ ELISA） method; the zonula occludens-1（ ZO-1）,Claudin-1,Occludin mRNA expressions were detected by real-time fluorescent quantitative polymerase chain raction（ q PCR） and ZO-1,Claudin-1 protein expressions were detected by immunohistochemistry method. Result： Compared with the normal group,the model group showed significant increases in serum DAO and TNF-α content in colonic mucosa（ P〈0. 01）,and decreases in ZO-1,Occludin,Claudin-1 mRNA expressions（ P〈0. 01） and ZO-1,Occludin protein expression（ P〈0. 01）; compared with the normal group,Shenling Baizhu particle groups showed decreases in serum DAO and TNF-α content in colonic mucosa（ P〈0. 01）, and up-regulations in ZO-1, Occludin, Claudin-1 mRNA expressions（ P〈0. 01） and ZO-1,Occludin protein expression P 0. 05,P〈0. 01） in functional diarrhea model rats. Conclusion： Shenling Baizhu particles can regulate tight junction protein expression in colonic mucosa,and intestinal mucosal mechanical barrier,which may be one of mechanisms of treating functional diarrhea.