整合序贯代谢与代谢组学的藏药小檗膏口服防治DR的组分协同研究

Angiogenesis mediated formation with vascular endothelial growth factor (VEGF) in diabetic retinopathy (DR) is a sign of worsening, concluding NSFC (81173360) found that Tibetan Medicine Berberidis Cortex Oral Extract obviously on the whole DR model, the mechanism related with regulation of the higher expression of VEGF on the whole, and that Tibetan medicine theory considered to balance ChiBa. This project intends to combine the drug absorption in vivo metabolism and "ChiBa in blood and perspiration" "eyes can see related with ChiBa in eyes" in Tibetan medicine theory, put forward the "The Berberidis Cortex Oral Extract prevention of DR related with the components collaborative mechanism into the blood to regulate VEGF on the whole, separation components of The Berberidis Cortex Oral Extract and carry out Component collaborative matrix to research, sequential metabolic for components in digestive enzyme metabolism, intestinal flora metabolism, intestinal mucosa metabolism, intestinal metabolism of liver, analysis methods such as UFLC-MS/FLD and etc. to find compositional variation, the overall identification of migrated into the blood of The Berberidis Cortex Oral Extract and its effective components and its metabolic site and its metabolic components; with the integration of sequential metabolism and Metabonomics, using spontaneous DR model to verify, molecular biology methods such as western blotting and etc. to detect the change and expression of P-gp and VEGF and other important facts. The principal component analysis method of mining the relationship with the metabolic and collaborative mechanism for components on prevention and treatment of DR, to guide the clinical medication of Tibetan medicine in further.

血管内皮生长因子VEGF介导的新生血管形成是糖尿病视网膜病变DR恶化的标志，结题NSFC81173360发现藏药小檗膏口服对整体DR模型作用明显，其机制与整体调控VEGF高表达有关，藏医认为与平衡赤巴增盛有关。本项目拟结合药物体内吸收代谢和“赤巴位于血汗内”“赤巴住眼能将诸物见”等藏医理论，提出“小檗膏口服防治DR与其代谢入血组分调控VEGF的协同效应有关”，分离小檗膏组分并构造组分协同矩阵，开展协同组分消化酶代谢、肠道菌群代谢、肠黏膜代谢、肠肝代谢等序贯代谢研究，UFLC-MS/FLD等手段分析组分变化特征，整体辨识小檗膏移行入血有效组分及其代谢成分；以自发性DR模型为验证，整合序贯代谢与代谢组学,免疫印迹等分子生物学手段，检测糖蛋白P-gp等肠吸收因子变化，检测血清及视网膜VEGF表达变化。主成分分析等方法挖掘其联系，揭示小檗膏口服防治DR的组分代谢与协同机制，进一步指导藏医临床用药。

血管内皮生长因子（VEGF）介导的新生血管形成是糖尿病视网膜病变（DR）恶化的标志，已结题NSFC81173360发现藏药小檗膏口服对整体DR模型作用明显，其机制与整体调控VEGF高表达有关，藏医认为与平衡赤巴增盛有关。本项目结合药物体内吸收代谢及序贯代谢等现代医学理论和“赤巴位于血汗内”“赤巴住眼能将诸物见”等藏医理论，在离体及在体实验中，发现盐酸小檗胺在肠吸收阶段、肠肝吸收阶段，盐酸小檗碱与盐酸小檗胺配比时，抗氧化活性几乎完全由盐酸小檗胺决定，且盐酸小檗胺可以促进盐酸小檗碱在肠肝中的吸收代谢，两者具有协同作用，确定了在小肠及肝微粒体中盐酸小檗碱与盐酸小檗胺的最佳配比组，为盐酸小檗碱的配比研究及制剂开发提供了科学依据；在db/db自发性糖尿病小鼠试验中，以小檗膏有效成分构建三因素六水平均匀设计矩阵，采用HPLC-MS技术，分析盐酸小檗碱、盐酸小檗胺、木兰花碱等入血成分，发现以序贯代谢的方式吸收入血的盐酸小檗碱部分代谢为盐酸小檗红碱，发现木兰花碱的给药剂量-血药浓度呈线性相关，各组别间存在差异的来源为盐酸小檗碱、盐酸小檗红碱，小檗膏有效成分在小檗碱-木兰花碱-小檗胺的配比能调节DR小鼠肠道菌群代谢、氨基酸代谢紊乱和脂质代谢紊乱，各组分相互协同具有改善DR病变特征，改善DR小鼠缺氧、炎症以及视网膜的新生血管形成，其中60/25/20（mg/kg）的剂量下具有最佳疗效，以小檗膏有效组分对糖尿病眼病的影响，这有利于揭示小檗膏口服防治DR的组分代谢与协同机制，进一步阐释了小檗膏口服防治DR的科学依据。本项目已培养毕业研究生4人，公开出版硕士研究生毕业论文3篇（均标注基金资助和项目编号），公开发表学术期刊论文13篇（均标注基金资助和项目编号，其中6篇已被SCI收录），项目负责人于2018年晋升教学科研型教授职称，项目组成员1人晋升副教授职称，荣获四川省高等教育优秀教学成果一等奖1次、四川省科技术技进步二等奖1次。