不同中医腹泻证型与肠道乳糖酶活性的相关性研究

The objective research of syndrome type in TCM (Traditional Chinese Medicine) is one of the important aspects of the TCM modernization where biosensor technology is playing a more and more important role by far. The intestinal lactase enzyme splits lactose into glucose and galactose, whose activity relates to diarrhea. We put forward a hypothesis about “there is a vital relationship between different diarrhea syndromes of TCM and the activity of the intestinal lactase enzyme, and the efficacy of TCM prescription in treating diarrhea relates to the activity of the lactase enzyme”. Using the principle and method of electrochemistry to develop a lactase biosensor, the project intends to establish a system to determine the activity of the intestinal lactase enzyme with a galactose biosensor. According to the classification of diarrhea in TCM field, using animal tests the project will study the changes of the lactase activity in intestinal between the two models that one is a different diarrhea syndromes of TCM , the other is a model after the intervention with the corresponding TCM prescription; and will investigate the relationship between different diarrhea syndromes of TCM and the activity of the intestinal lactase enzyme; reveal the characteristics of the lactase activity in different diarrhea syndromes and the mechanisms of TCM prescription in treating diarrhea. In the results we will establish a biosensor detection technology system for the objective diagnosing method of different diarrhea syndromes of TCM, and provide an objective index system for the diagnosis of different diarrhea syndromes of TCM. It can present a novel method and some new ideas for the objective diagnosis and treatment of TCM based on the relating function enzymes of disease. The contributions of the project will have great significance to the digital clinical diagnosis and modernization of TCM.

中医证型客观化研究是中医现代化的重要方面，生物传感测量技术在中医药研究领域的优势日渐凸显。肠道乳糖酶水解乳糖为葡萄糖和半乳糖，其活性与泄泻具有相关性。我们提出“不同中医泄泻证型与肠道乳糖酶活性具有相关性，中医方剂对泄泻的疗效与肠道乳糖酶活性有关”的假说。本项目拟运用电化学的原理，研制半乳糖传感器，建立肠道乳糖酶活性检测的生物传感检测技术体系；根据中医对泄泻的分型，通过动物试验，研究不同中医泄泻证型造模及相应中医方剂干预后肠道乳糖酶活性的变化，探讨肠道乳糖酶活性与中医泄泻证型的相关性，揭示不同泄泻证型乳糖酶活性的特征，探究中医方剂治疗不同中医泄泻证型的机理。研究结果可建立生物传感检测技术用于中医泄泻证型的客观化诊断，提供中医泄泻证型诊断的客观化指标体系，可为疾病相关功能酶用于建立中医证型客观化诊疗技术提供新的方法和思路，对中医临床诊断数字化及现代化意义重大。

中医证型客观化研究是中医现代化的重要方面。肠道乳糖酶活性与泄泻具有相关性。围绕本项目进行的主要研究及结果如下：.1）运用“高糖高脂+高温高湿+饮酒+冰水”法成功制备了肠道湿热证泄泻小鼠模型，模型动物肠黏膜中乳糖酶、蔗糖酶活性显著降低（P < 0.05）。葛根芩连汤对肠道湿热证泄泻小鼠疗效确切，提高了肠道内容物和肠黏膜中总乳糖酶、蔗糖酶活性（P < 0.05），且随治疗时间的延长，肠前段黏膜乳糖酶和蔗糖酶活性逐渐恢复而后段黏膜乳糖酶和蔗糖酶活性升高。以上表明，肠道湿热证泄泻的发生使肠黏膜乳糖酶和蔗糖酶活性下降，葛根芩连汤疗效的发挥与其调节肠道乳糖酶和蔗糖活性相关。. 2）采用“番泻叶+束缚+夹尾”法成功复制了肝气乘脾泄泻小鼠模型。模型动物甘油三酯（TG）、低密度脂蛋白胆固醇（LDL-C）、5-HT含量升高（P＞0.05），D-木糖含量下降（P＜0.05），肠胃功能亢进。痛泻要方对模型小鼠疗效显著，降低了小鼠总胆固醇（TC）、高密度脂蛋白胆固醇（HDL-C）（P＜0.05）、TG、LDL-C含量（P＞0.05），抑制胃肠功能亢进，使小鼠肠前段黏膜乳糖酶和蔗糖酶活有所回归，调节中段黏膜乳糖酶活性恢复，显著提高后段黏膜乳糖酶、蔗糖酶活（P＜0.001）。以上表明，痛泻要方对肝气乘脾泄泻小鼠不同肠段黏膜乳糖酶和蔗糖酶活性作用显著，为肝气乘脾泄泻的发生及治疗研究提供了理论基础。.3）采用硫酸庆大霉素和头孢拉定制备抗生素相关性腹泻小鼠模型（属寒湿泄泻范畴），小鼠肠道黏膜乳糖酶活性降低，七味白术散对腹泻小鼠不同肠段黏膜乳糖酶活性具有恢复或提高作用。研究结果对于临床抗生素相关性腹泻的诊治具有重要参考意义。.4）研究了ONPG法和双糖酶法对乳糖酶活性测定的线性范围、检出限、稳定性、灵敏度，结果表明ONPG法优于双糖酶法用于肠道乳糖酶活性测定。