从平胃散干预前后水通道蛋白0(AQP0)的定性定量表达研究湿阻中焦证的水液代谢机制

目的观察并分析水通道蛋白0(AQP0)在大鼠消化段(胃贲门、胃体中部、大肠中段、大肠末段)的差异性表达,推测其与消化道生理功能的相关性及平胃散对消化道水液代谢的干预机制。方法通过对三大致湿因素的联合应用(外湿过盛,碍胃困脾;饮食不节,内生湿邪;情志不遂,水湿失布),建立湿困中焦的动物病理模型,采用免疫组织化学技术检测AQP0在消化段的分布及表达情况。结果 AQP0主要在消化道的黏膜层表达,黏膜下层、肌层和外膜处可有疑似表达。空白组与模型组相较而言:AQP0在胃贲门黏膜层表达有增加的趋势。自然恢复3 d后,相较于模型组:AQP0在胃贲门黏膜层,大、小肠中段表达有增加的趋势,胃体中间有降低的趋势。经平胃散干预3 d后,相较于模型组:AQP0在胃贲门和大肠中段黏膜层有增加的趋势,胃体中部和小肠中段黏膜的表达略有降低。且平胃散干预组与自然恢复组比较,AQP0在胃贲门、胃体中部、小肠中段黏膜层的表达有所降低。同时,空白给药组相较于空白组而言:AQP0在消化道各段黏膜层的分布有增加的趋势。结论①AQP0在消化道黏膜层表达,可能与水分吸收,腺体分泌的调控机制相关;②AQP0在胃和肠的表达有差异,内环境pH可能是影响因素之一;③平胃散能促进AQPO的表达,可以通过AQP0维持水液代谢的平衡,这可能是平胃散燥湿健脾、散满和胃在分子水平的机制之一。

A Study on Expressional Distribution Spectrum of the Pathological Features of Aquaporin0(AQP0)in An Animal Model on the Syndrome of Dampness Incumbering Middle Energizer in Gastrointestinal Tissue and the Intervention by Pingwei Dosage

Objective To observe and analyze the different expression of water channel protein 0(AQP0) in rat’s digestive tract(Cardia of stomach、the middle of stomach、the middle of small intestine、the middle of large intestine 、the end of large intestine),speculate the relevance between AQP0 and the physiologic function about alimentary canal and the mechanism about the pathogenesis in disturbance of water metabolism caused by Pingwei Dosage. Methods United three major pathogenic factors(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).Build a model of the syndrome of dampness incumbering middle energizer.Measure the distribution of the AQP0 in the gastrointestional tissue by immunohistochemistry. Results Expressional content of AQP0 concentrated in mucosa of the alimentary canal,the expression in other parts were unsure.Compared with the model team,expressional content of AQP0 in blank team enhanced in mucosa tissue of the cardiac stomach.After three day’s renew,expressional content of AQP0 enhanced in mucosa tissue of the cardiac stomach,the middle of small/large intestine,but reduced in the middle of the stomach.Pingwei dosage enhanced the expressional content in mucosa tissue of the cardiac stomach and the middle of large intestine,reduced little in middle part of the stomach and small intestine.Compared with the renew team,using Pingwei dosage could reduce the expressional content in mucosa tissue of the cardiac stomach 、the middle of the stomach and small intestine.compared with blank team,using Pingwei dosage could enhance the expressional content in mucosa tissue in the whole alimentary canal. Conclusion (1)AQP0 that expressed in gastrointestional tissue may be one of the molecular basis in moisture absorption、glandular secretion.(2)The expression contents of AQP0 are different between the stomach and the intestine,It maybe caused by the distinct Ph.(3) Pingwei dosage can enhance the expression of AQP0 and maintain the balance of the metabolic of water by AQP0,maybe it’s one of the reason which can explain Pingwei dosage’s efficacy on eliminating dampness to activate spleen,scattering stagnant to comfort stomach.