肝郁脾虚型肠易激综合征的肠菌群代谢及痛泻要方的干预

Irritable bowel syndrome ( IBS ) is a common functional gastrointestinal disease in adults. The intestinal microflora composition and metabolic of IBS patients is different from healthy control people. Researchs suggest that gut dysbiosis is the fundament of IBS to part patients. TCM spleen deficiency patients have intestinal dysbacteriosis usually, and we observed a similar situation of dysbiosis in liver stagnation and spleen deficiency syndrome animal model of IBS. Treatment of IBS with traditional Chinese medicine is super to western medicine, the mechanism of TCM base on intestinal flora metabolics has no in-depth study. " Tongxie Yaofang" is a classic prescription of IBS,among the prescription, Largehead atractylodes rhizome(Bai Zhu) is used to treat spleen deficiency syndrome. Did " Tongxie Yaofang" have a recovery effect to the imbalance of intestinal flora? The project uses the metabonomics to intestinal flora of IBS rat model with liver stagnation and spleen deficiency syndrome, the intestinal contents of rat are extracted and derivatization, detected by gas chromatography mass spectrometry. The metabolite profiling will be established and the features clarified; The 16SrDNA fingerprint of intestinal microflora is constructed by DGGE technology, the construction of intestinal flora in different state is compared. " Tongxie Yaofang " is inducted to the research mode compared with other drugs` effect, and the intestinal bacterial metabolism characteristics and composition changes are observed before and after administration. The project is going to establish a metabolic modle to IBS based on of TCM description and a judge standard of the curative effect.

肠易激综合征(IBS)是常见于青壮年的功能性肠道疾病。IBS患者肠道菌群构成和代谢有异于常人，研究认为肠道微生态失调是部分IBS患者的基础病因。中医的脾虚患者多有肠道菌群失调，我们在前期制作肝郁脾虚型IBS动物模型时也观察到类似情况。IBS是中医药治疗的优势病种，其基于中医辨证分型的肠道菌群代谢尚无深入研究。含有健脾药白术的"痛泻要方"是治疗肝郁脾虚型IBS经典方，该方是否通过恢复失调的肠道菌群产生部分疗效？本项目利用代谢组学方法研究肝郁脾虚型IBS模型大鼠的肠道菌群，将大鼠肠内容物提取并衍生化，以气相－质谱检测，建立代谢物谱，明确其特征；用DGGE技术构建肠菌群16SrDNA指纹图谱，考察不同状态下肠内菌群构成；并将疏肝扶脾的"痛泻要方"切入到这一模式中，对比其他药物，观察给药前后肠道细菌代谢特征和构成变化，以期建立一个基于中医辨证分型的IBS代谢组学描述模式，并作为疗效判断的一个标准。

肠易激综合征(IBS)是常见于青壮年的功能性肠道疾病。IBS 患者肠道菌群构成和代谢有异于常人，研究认为肠道微生态失调是部分IBS 患者的基础病因。中医的脾虚患者多有肠道菌群失调，我们在前期制作肝郁脾虚型IBS 动物模型时也观察到类似情况，进而认为中医某些分型的IBS患者或类似症候的动物模型可能具有一定的菌群结构特征和代谢倾向。. 项目利用中药指纹图谱对痛泻要方及造模用的中药进行质量监控，确保每批次提取药物的有效成分基本一致。用DGGE 技术考察肝郁脾虚型IBS 动物模型的肠内容物菌群结构，构建肠菌群16SrDNA 指纹图谱，考察不同状态下肠内菌群结构变化，同时观察痛泻要方对菌群结构的作用。发现对肝郁脾虚型IBS模型大鼠进行痛泻要方和阳性止泻药（匹维溴铵）的治疗后，提取各组的新鲜粪便总DNA，检测扩增后DGGE指纹图谱。对各组条带反映的生物多样性进行分析，模型组的生物多样性（H,仙农指数）明显降低，而痛泻要方两个剂量对其均有恢复作用，同时物种的丰度(S)也有明显改善。进一步对菌群构成进行聚类分析，发现痛泻要方干预后的个体在分类树中均很靠近。提示痛泻要方可以使肠道菌群结构更为稳健，且其促进肠菌群增殖有一定的倾向性。 肠道内容物样本经氯甲酸丙酯衍生化后进行气相-质谱（GC/MS）检测代谢谱的变化。结果表明肝郁脾虚型IBS大鼠的粪便SCFA中的乙酸含量较高，给予痛泻要方治疗后乙酸含量降低，在治疗前后有显著差别。. 项目对IBS的基础病因进行探讨，验证了符合中医特定分型的动物模型与正常动物的肠内容物在菌群结构方面存在差别。针对这种中医症候分型的药物（痛泻要方）能较好的恢复其菌群结构，并降低其代谢物中乙酸的含量。项目从菌群结构和代谢组学角度验证了痛泻要方对肝郁脾虚型IBS的方证对应，并对其特征进行考察，为进一步的药物和IBS病理机制研究打下基础。