基于肠道乳糖酶对七味白术散治疗小鼠菌群失调腹泻的机理研究

腹泻是许多疾病的常见症状。七味白术散对腹泻疗效显著。肠道酶来自动物及微生物，乳糖酶缺乏与多种腹泻相关。前期研究发现，抗生素造模降低小鼠对乳糖的代谢，七味白术散提高菌群失调腹泻小鼠对乳糖的代谢，其对乳糖酶的影响与机理未见报道。本项目拟运用抗生素制备菌群失调腹泻小鼠，灌胃七味白术散，分别采集肠道内容物及粘膜，将酶学、分子生物学和宏基因组技术结合，分析乳糖酶活性、（小鼠、细菌及真菌）乳糖酶基因多样性、（小鼠、细菌及真菌）乳糖酶基因表达量，从酶活、基因及基因表达等探析七味白术散对乳糖酶的影响机制。结合七味白术散对菌群失调腹泻的疗效，揭示七味白术散治疗菌群失调腹泻的机理。项目结果为利用乳糖酶阐明七味白术散治疗菌群失调腹泻的机理提供实验依据。项目研究成果与技术体系，对利用疾病相关重要肠道酶建立中医药辨证论治标准化诊疗技术及阐明中医药疗效机理意义重大。

Diarrhea is a common clinical symptom of many diseases. Qiweibaizhusan has a great effect on curing diarrhea. Intestinal enzymes, which are mainly secreted from animal and microbes, have significant impacts on the transformation functions in the gastrointestinal tract. Insufficiency of the intestinal lactase, which hydrolyzes lactose, is associated with many kinds of diarrhea. Our previous studies showed that lactose metabolism in mice was reduced by mixed antibiotics modeling. And lactose metabolism in mice with dysbacteriosis diarrhea was improved by Qiweibaizhusan. But the adjustment function and mechanisms of Qiweibaizhusan on intestinal lactase are not investigated. In order to investigate the curative mechanisms of Qiweibaizhusan upon dysbacteriosis diarrhea, this project will use enzymological techniques and molecular biological techniques combining with metagenome technology. The mouse model with dysbacteriosis diarrhea will be established by administrating mixed antibiotics, and then treated with Qiweibaizhusan. Intestinal content and mucosa will be collected respectively. The changes of intestinal lactase activity in the tested mice will be detected. Gene polymorphisms of the lactase from mice, bacteria and fungi will be analysed, respectively. The gene expression level of lactase from mice, bacteria and fungi will be measured, respectively. The relevance of intestinal lactase activity, gene polymorphism of lactase, and gene expression of lactase will be determined. The relationship between the curative effects of Qiweibaizhusan upon dysbacteriosis diarrhea and the intestinal lactase including activity, gene diversity and gene expession in the tested mice will be analysed. Outcomes of the project will provide experimental basis by using lactase to elucidate curative mechanisms of Qiweibaizhusan upon dysbacteriosis diarrhea. The research findings and methodologies may have a great significance in using disease related important gastrointestinal enzyme for establishing standardization technology of Chinese medicine diagnosis and treatment based on syndrome differentiation and elucidating curative mechanism of traditional Chinese medicine.