脾肾阳虚证腹泻型肠易激综合征大鼠模型及其生物学基础的研究

Diarrhea predominate irritable bowel syndrome (IBS-D) is one of the most common disease among the digestive system diseases and Traditional Chinese Medicine (TCM) is efficacious to IBS-D. Spleen kidney Yang deficiency syndrome (SKYDS) is the main syndrome of IBS-D and warming the kidney and strengthening the spleen (WKSS) therapy showed a satisfied curative effect in clinic. Syndrome differentiation and treatment is the basic principles of understanding and treating diseases. Chronis stress is the classic method to build the IBS-D model. In order to further study the mechanisms of TCM in treating IBS-D with SKYDS, an IBS-D model with SKYDS is urgently required. Our previous study demonstrated that different dose and duration of Senna oral administrations could lead to different manifestations of SKYDS. In this study, we plan using chronic stress combining with Senna oral administration to build the IBS-D model with SKYDS, and a four-dimensional evaluation system, including behaviors, biomarkers, medicine counterevidence and building factors backstepping, is planning to evaluate the model. Thus, this study will provide a stable and repeatable model, filling the academic gap. In addition, metabonomics and intestinal microflora are used as the breakthrough point, not only screening the potential biomarkers, to further enrich the evaluation system, but also searching the potential targets of TCM in treating IBS-D, laying the foundation of mechanisms researches of TCM in treating IBS-D.

腹泻型肠易激综合征（IBS-D）是中医治疗优势病种，脾肾阳虚证为其主要证型之一，温肾健脾法临床疗效显著。辨证论治是中医认识及治疗疾病的基本原则，目前慢性应激法是IBS-D的经典造模方法，为深入研究中医药作用机制，构建公认的脾肾阳虚证IBS-D动物模型势在必行。团队前期研究发现不同剂量、周期番泻叶灌胃可导致大鼠不同程度脾肾阳虚表现。本研究拟在此基础上采用慢性应激联合番泻叶灌胃建立脾肾阳虚证IBS-D模型，按照“宏观体征-理化指标-药物反证-造模因素反推”四维评价体系进行模型评价，为业内提供稳定、可重复的脾肾阳虚证IBS-D模型，填补目前研究空白。并以代谢组学和肠道微生态为切入点，进一步丰富模型的评价体系，挖掘中药潜在作用靶点，为中医药治疗IBS-D机制研究奠定基础。

背景：腹泻型肠易激综合征（IBS-D）是中医治疗优势病种，脾肾阳虚证为其主要证型之一，构建公认的脾肾阳虚证IBS-D动物模型势在必行。目的：采用慢性应激联合番泻叶灌胃建立脾肾阳虚证IBS-D模型，按照“宏观体征-理化指标-药物反证-造模因素反推”四维评价体系进行模型评价。方法：①母子分离法建立IBS疾病模型。以高、中、低番泻叶浓度（1.0g/ml、0.5g/ml、0.25g/ml）灌胃，探索构建脾肾阳虚证所需的剂量和周期。②实验动物共计210只，分为母子分离组+番泻叶高、中、低剂量（灌胃2、4、6周）；直肠扩张+番泻叶高、中、低剂量（灌胃2、4、6周）；避水应激+番泻叶高、中、低剂量（灌胃2、4、6周）；正常组（灌胃的2、4、6周）。共30组。比较优化出最佳的病证结合动物模型。③以母子分离+番泻叶灌胃造模，35只大鼠分为正常组、模型组、温中健脾组、温补肾阳组、温肾健脾组各7只。予相应治疗后对各指标进行评价。④代谢组学分析实验一模型构建过程中大鼠尿液代谢轮廓和轨迹变化，筛出差异代谢物。⑤从肠道微生态角度探索实验一模型组和正常组的菌群差异。结果：①高剂量番泻叶灌胃出现大鼠体重增长减缓、进食减少、握力降低和负重游泳时间减少、肛温降低、尿-17羟和CORT降低（P<0.05），最佳灌胃时间为6周。②直肠扩张和母子分离联合高剂量番泻叶灌胃6周均能稳定的构建脾肾阳虚证IBS-D大鼠模型。③温肾健脾组疗效优于温中健脾组及温补肾阳组。④模型组大鼠上调的差异代谢物主要有肌酸、尿囊素等6个，下调的差异代谢物主要有乙醇、酮戊二酸等5个。⑤模型组大鼠呈现厚壁菌门增加，拟杆菌门减少趋势。结论：①构建脾肾阳虚证IBS-D大鼠模型最佳剂量为高剂量组，最佳灌胃周期为6周。②直肠扩张和母子分离联合高剂量番泻叶灌胃6周均能稳定的构建脾肾阳虚证IBS-D大鼠模型。③证明前期建立的动物模型符合中医病证结合特点。④模型组大鼠在能量代谢、应激相关代谢、肠道菌群、胆汁酸代谢发生了代谢信息改变，肠道菌群的菌群结构亦发生改变。