荷瘤大鼠肠道菌群对白花蛇舌草黄酮苷的代谢转化及其多糖对该代谢的影响研究

The study on glucosides' Intestinal flora metabolic transformation has been always carried out in normal condition bodies, without considering the effect of disease states. The flavonoid glycoside and polysaccharide, which could inhibit or kill the tumor directly and enhance the immune function respectively, were the main anti tumor active ingredients in Oldenlandia diffusa. With the Intestinal flora's abnormal metabolism in disease states as the starting point of the research on Oldenlandia diffusa flavonoid glycoside's Intestinal flora metabolic transformation , we came up with this hypothesis: the disease states destroied the balance in Intestinal flora and impacted the metabolic transformation; the Oldenlandia diffusa polysaccharide, which could regulate the unbalanced intestinal flora to normal and improve and recover the metabolic transformation fuction for flavonoid glycoside by stimulating the intestinal mucosa immune response, supplying the needed sugar, recovering the quantity and vitality of intestinal probiotics. We planned to use tumor-bearing animal models, combining HPLC-MS, to compare the differences of Oldenlandia diffusa flavonoid glycoside's Intestinal flora metabolic transformation between normal conditions and diseases, and to investigate the regulation fuction of Oldenlandia diffusa polysaccharide in this transformation. Discuss the effect on intestinal flora' s metabolic transformation fuction for aminoglycoside drugs under disease states, and the regulation of polysaccharide in this metabolic transformation, prove the traditional Chinese medicine had such function characteristics as multicomponent, multi-targets and multiple pathways, thus provide new ideas for study on the aminoglycoside drugs' intestinal flora metabolic transformation.

糖苷类成分的肠道菌群代谢转化研究均在机体正常状态下开展，未考虑疾病状态的影响。白花蛇舌草中的黄酮苷和多糖是其主要的抗肿瘤活性成分，分别具有直接抑杀和免疫调节功能。以疾病状态（荷瘤状态）下机体肠道菌群的异常作为研究白花蛇舌草黄酮苷肠道菌群代谢转化的切入点，并提出假说：疾病状态（荷瘤状态）破坏了肠道菌群平衡，影响黄酮苷的代谢转化；应用白花蛇舌草多糖，可刺激肠道粘膜免疫应答，提供肠道菌群所需糖源，恢复肠道益生菌数量和活力，从而调整失衡的肠道菌群趋于正常，提高和恢复对黄酮苷的代谢转化能力。拟采用荷瘤动物模型结合HPLC-MS技术，比较白花蛇舌草黄酮苷在正常及荷瘤状态下肠道菌群代谢转化的差异，考察白花蛇舌草多糖对该代谢转化的调整作用。探讨疾病状态下肠道菌群对糖苷类药物代谢转化功能的影响及多糖对该代谢的调整，揭示中药多成分、多靶点、多途径的作用特点，为糖苷类药物的肠道菌群代谢转化研究提供新思路。

糖苷类成分代谢受机体诸因素影响，疾病状态下机体生理功能紊乱从而影响其代谢过程。黄酮苷类成分是抗肿瘤中药白花蛇舌草活性物质之一，目前其研究多集中于正常机体的肠道菌群转化，未考虑疾病状态影响。中药多糖类成分作为肠道菌群所需糖源，可调节及修复肠道菌群谱。白花蛇舌草多糖具有免疫调节作用，同时亦为抗肿瘤活性物质。本项目以疾病状态下机体肠道菌群异常作为切入点，并提出假说：疾病状态（荷瘤状态）下肠道菌群紊乱而影响黄酮苷的代谢转化；通过白花蛇舌草多糖对肠道菌群的调节修复，可提高对黄酮苷的代谢转化能力。.项目组首先对白花蛇舌草黄酮苷及总多糖进行纯化制备及定量分析；通过考察黄酮苷人工胃、肠液的稳定性和大鼠离体肠吸收试验，确立其代谢转化部位；同时采用Caco-2细胞模型，揭示黄酮苷及苷元在体内吸收特点；通过黄酮苷体外肠道菌群转化试验，确证苷元为其肠道菌群代谢产物。在此基础上，构建SD荷瘤大鼠模型，比较白花蛇舌草黄酮苷在正常及荷瘤状态下肠道菌群代谢转化的差异，并通过illumina MiSeq 2 x 300 bp 高通量测序平台对白花蛇舌草多糖调节肠道菌群测序，研究多糖对肠道菌群的调节修复作用及对黄酮苷体内代谢转化的影响。.研究表明，荷瘤大鼠黄酮苷药代动力学特征异于正常机体状态，其经多糖调节后的药代动力学特征相比于荷瘤状态有所改善；肠道菌群测序结果显示，荷瘤状态下肠道菌群在各水平均有不同程度变化，经多糖调节后对肠道菌群具有一定修复作用，有效证实了本假说的科学性。.本项目对黄酮苷在疾病状态下的代谢转化进行研究，并通过多糖对肠道菌群的调节作用，研究其对代谢转化的影响，揭示中药多成分、多靶点、多途径的作用特点，为糖苷类药物的肠道菌群代谢研究提供新思路。