湿阻中焦证模型胃肠水通道蛋白1的病理特征性表达分布谱以及平胃散干预的研究

A Study on Expressional Distribution of the Pathological Features of Aquaporin1(AQP1)in a Animal Model on the Syndrome of Dampness Incumbering Middle Energizer in Gastrointestinal Tissue and the Intervention by Pingwei Powder

目的:本实验以前期较成熟湿阻中焦证动物模型为研究平台,研究湿阻中焦证动物模型胃肠组织AQP1的分布和含量以及平胃散的干预,揭示湿阻中焦证动物模型水通道蛋白病理特征性表达分布谱。从蛋白水平揭示湿阻中焦证在水液代谢失调病机方面以及平胃散治疗湿阻中焦证的科学内涵;搭建中药复方水通道调节剂的研究平台。方法:模拟外湿过盛,困阻脾胃、饮食不节,湿从内生、情志不遂,气机受阻,水湿不运三大致湿病因,建立湿阻中焦证模型。用免疫组化技术测定胃肠组织AQP1的分布,用酶联免疫法(ELISA)测定胃肠组织AQP1的含量。结果:湿阻中焦证AQP1在胃贲门黏膜下组织和胃体中间黏膜表达减少,AQP1在胃贲门和小肠中段黏膜表达增加。平胃散增强湿阻中焦证胃贲门黏膜下组织、胃体中间黏膜和小肠中段黏膜AQP1的表达,抑制湿阻中焦证胃贲门黏膜AQP1的表达。结论:AQP1在湿阻中焦证胃肠特征性表达分布谱可能是湿阻中焦证湿邪困阻脾胃,阻遏气机,散输水津失调所表现证候的分子基础之一。平胃散对湿阻中焦证胃肠AQP1表达的干预可能是平胃散燥湿运脾、行气导滞、散中焦之湿阻治疗湿阻中焦证的分子机理之一。平胃散增强正常大鼠胃肠AQP1的表达,可能是平胃散苦燥虽可祛湿但也可伤津液的分子机理之一。平胃散治疗湿阻中焦证的疾病临床疗效确切,但也不能乱服多服。

Objective：Based on the animal model of the syndrome of Dampness Incumbering Middle Energizer,to study the distribution and content of the aquaporin 1（AQP1） in the model,to reveal its expressional distribution spectrum of the pathological features of aquaporin.To study syndrome mechanism caused by the pathogenesis in disturbance of water metabolism and scientific intension of treatment by Pingwei Powder in the model in protein level,to build a research platform of aquaporin conditioning agent with traditional Chinese medicine formular.Methods：Build a model of the Syndrome of dampness Incumbering Middle Energizer,simulating the three big etiological factors of retention of Water and dampness：i.e.the functioning declination of Spleen and Stomach by exterior Dampness luxus,interior Dampness caused by intemperant diet,or stagnation of Qi caused by emotional depression.Measure the distribution of the AQP1 by Immunohistochemistry and the content of the AQP1 by enzyme-linked immuno sorbent assay（ELISA） in the gastrointestional tissue.Results：In the model of the Syndrome of Dampness Incumbering Middle Energizer,expressional content of AQP1 reduced in submucous tissue of the cardiac stomach and in mucosa of mid-corpusventriculare,while AQP1 increased in mucosa of cardiac stomach and middle part of small intestine.Pingwei Powder enhanced the expressional content of AQP1 in submucous tissue of the cardiac stomach,mucosa in mid-corpusventriculare and mucosa in middle piece of small intestine;depressing the content of AQP1 in mucosa of cardiac stomach in the model.Conclusion：Expressional distribution spectrum of the pathological features of AQP1 in the gastrointestinal tissue of the animal model of the Syndrome of Dampness Incumbering Middle Energizer may be one of the molecular basis in this syndrome caused by disorder of water and fluid,which resulting in function declination of spleen and stomach,and stagnancy of Qi-movement.The expressed intervention on distribution of AQP1 by Pingwei Powder in the model may be one of the molecular basis that Pingwei Powder could eliminate dampness to activate spleen,promote Qi to remove stagnancy,and expel dampness in Middle Energizer,to cure the syndrome.The enhancement in the expressed distribution of AQP1 in normal rats＇ gastrointestinal tract enhanced by Pingwei Powder,may be one of the molecular basis,that the bitter and dry nature of Pingwei Powder can drive out dampness,but may also impairs body fluids.Pingwei Powder has positive clinic effects on the Syndrome of Dampness Incumbering Middle Energizer,but cannot be overtaken or abused.