论文部分内容阅读
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 observe the effect of Shenling Baizhu Granule on the intestinal mucosal tight junction protein in rats with functional diarrhea and its mechanism. Methods: A rat model with functional diarrhea and spleen deficiency syndrome was established by legislation of high lactose + small platform. 90 rats were randomly divided into normal group, model group (high lactose + microtiter plate), piroxicam group, The high, medium and low dose groups of 15, each group, Shenling Baizhu granules high, medium and low doses were 3.212,1.606,0.803 g · kg ~ (-1)), 0.012 5 g · kg ~ (-1)). Normal group and model group were fed with normal saline 3 times a day for 14 days. Serum levels of diamine oxidase (DAO), tumor necrosis factor-α (TNF-α), real-time fluorescence quantitative polymerase chain reaction (qon), zonula occludens ~ (-1), ZO ~ (-1)), and the blocking protein ~ (-1) were detected by q PCR. Claudin ~ (-1)) and Occludin mRNA were detected by immunohistochemical method. The expression of ZO ~ (-1) and Occludin was detected by immunohistochemistry. Results: Compared with normal group, serum levels of DAO and TNF-α in colonic mucosa of model group were significantly increased (P <0.01), ZO -1, Claudin ~ (-1), Occludin (P <0.01), and the expression of ZO ~ (-1) and Occludin was significantly decreased (P <0.05). Compared with the model group, Shenling Baizhu Granule could reduce the level of DAO in serum, (-1), Claudin ~ (~ (-1)) and Occludin mRNA (P <0.01) in upregulated ZO ~ (-1) (~ (-1)), Occludin protein expression (P <0.05, P <0.01). Conclusion: Shenling Baizhu Granule can regulate the colonic mucosal tight junction protein expression and regulate the mechanical barrier of intestinal mucosa in functional diarrhea rats, which may be one of the mechanisms of treating functional diarrhea from spleen.