京大戟醋制减毒的效应物质基础和作用机制研究

As a common drastic diuresis medicine of TCM clinical, Euphorbiae Pekinensis Radix, the toxic Chinese materia medica must be stir-baked with vinegar for oral administration (OA). However, the existing research results not yet been able to fully answer the scientific question, namely, the material basis and action mechanism of vinegar processing for reducing toxicity of Euphorbiae Pekinensis Radix. According to the theory of Euphorbiae Pekinensis Radix particular medicine property and Chinese medicine clinical practice of detoxifying, reducing toxicity and abating medicine property in the clinical practice, also the preliminary interpretation of the mechanism study of vinegar processing for reducing gastrointestinal toxicity of Euphorbiae Pekinensis Radix, we proposed a hypothesis based on the previous studies about that: the toxicity reduction and effect retention after processing Euphorbiae Pekinensis Radix with vinegar might be achieved by the chemical conversion and in vivo metabolism, which can reduce Casbane diterpene toxicity. This item plans to select several biological indicators which can reflect the characteristics of Traditional Chinese Medicine(TCM) syndrome medication, combined with the application of bio-guided separation technology to screen the active substances and explore the correlation between material basis and effection on the crude and processed Euphorbiae Pekinensis Radix. To reveal the intrinsic nature of vinegar processing for reducing toxicity by studying the differences in the toxicity action mechanism of Euphorbia Pekinensis Radix processing with vinegar under cells, normal animals and pathological model from the different levels of whole, organs, cells and molecules and the characteristics of model animal in vivo metabolic. This study will provide scientific basis for the clinical use of Euphorbiae Pekinensis medicines, and explore the methods for the study of the clinical use of toxic medicines.

有毒中药京大戟为中医临床常用峻下逐水药，内服需醋制。但现有研究结果尚未能全面解答京大戟醋制减毒的物质基础和作用机制等科学问题。基于对京大戟独特药性理论的深刻认识和中医临床采用醋制以解毒降毒、缓和药性的实践及课题组前期对醋制降低京大戟胃肠毒性作用机制的初步阐释，提出：京大戟醋制减毒作用可能是通过化学转化和体内代谢改变了毒性物质Casbane型二萜类化合物的结构生成低/无毒的效应物质而实现的。因此本研究选择符合京大戟中医辨证用药特点的生物效应评价指标，应用生物导向分离技术追踪京大戟醋制前后的效应物质，寻找京大戟醋制减毒物质基础-效应相关性。采用细胞、正常动物和病理模型，从整体、组织、细胞及分子不同水平分析京大戟醋制前后毒性作用机制的差异及在模型动物上体内代谢组学的特点，以期揭示京大戟醋制减毒的作用机制，为京大戟临床安全有效应用提供科学依据，为有毒中药科学应用基础研究提供思路与方法等方面的探索。

有毒中药京大戟为中医临床常用峻下逐水药，内服需醋制。但现有研究结果尚不能全面解答京大戟醋制减毒的物质基础和作用机制等科学问题。基于对京大戟独特药性理论的深刻认识和中医临床采用醋制减毒、缓和药性的实践及课题组前期对醋制降低京大戟胃肠毒性作用机制的初步阐释，从以下方面进行京大戟醋制减毒的效应物质基础和作用机制研究。选择符合京大戟中医辨证用药特点的生物效应评价指标，以正常小鼠、斑马鱼（胚胎）、胃肠上皮细胞为研究对象，确定了京大戟醋制减毒的有效部位为石油醚-乙酸乙酯部位。应用生物导向分离得到京大戟中17个化合物，并追踪其中6个效应物质成分的含量在醋制后都有所降低。以正常小鼠为研究对象，京大戟对肝胃肠均有氧化损伤作用，醋制可降低对小鼠肝胃肠的氧化损伤；京大戟对斑马鱼胚胎的肝胃肠器官均有毒性，醋制可降低京大戟不同极性部位对斑马鱼胚胎的肝、胃肠毒性。京大戟中casbase型二萜类化合物pekinenal和pekinenin C均有显著的肠上皮细胞毒性，能够影响IEC-6细胞周期进程，诱导细胞凋亡，并且均能从线粒体路径和死亡受体路径诱导肠上皮细胞凋亡。考察了癌性腹水大鼠模型组及各给药组对大鼠尿液、血清、肝、胃、肠组织生化指标及氧化损伤指标的影响，发现生、醋京大戟均有通利二便而泻水逐饮的作用和改善氧化损伤的作用。利用已建立的癌性腹水大鼠模型进行尿液和血液代谢组学研究，鉴别出血清中15个潜在生物标志物，参与了11条体内代谢路径；尿液中10个潜在生物标志物，参与了7条体内代谢路径，两者存在较大的差异，但生、醋京大戟均可通过调节相应代谢通路而对模型产生干预作用。通过上述研究可知京大戟对肝胃肠均有毒效作用，其中对肠道更为明显，这些为京大戟临床安全有效应用提供科学依据，为有毒中药科学应用基础研究提供思路与方法等方面的探索。