基于细胞微环境对《内经》“有故无殒”的现代生物学机制研究

You Gu Wu Yun documented in Nei Jing is an important cognition to the traditional toxicity theory of TCM, which mearns that a drug will reveal its therapeutic effect when it is prescribed to patients with the correct indications; however, it may produce deleterious effects in both sick and healthy people as a result of incorrect indications. The You Gu Wu Yun theory devotes important theoretical and practical values in the toxicity assessment and rational use of TCM drugs. However, the biological mechanism of You Gu Wu Yun is in lack of scientific clarification to date. We propose in this item that the macroscopic diversity existed in respond to toxicity of drugs may be caused by the difference between the microenvironments of both pathological and physiological statuses, especially for immune microenvironment in microscopic view, which has been preliminarily proved to be rational in our former research and the literatures. To do this, the liver toxicities of TCM drugs containing anthraquinones such as rhubarb and Polygonum multiflorum were studied in focus in this project. The immunology methods will be employed to screen the key regulators of liver microenvironment related to You Gu Wu Yun phenomenon of rhubarb. The research will provide new vision and illustration of the modern biological mechanism of the ancient TCM toxicity theory You Gu Wu Yun, as well the scientific evidences for the hepatotoxicity assessment and rational use of those drugs containing anthraquinones.

《内经》"有故无殒"是最能反映中医辨证用药思想的理论认识之一，通俗地说即是"有病则病当之，无病则体受之"，对指导中药毒性评价与合理用药具有重要的理论和实际价值。然而，"有故无殒"的生物学机制和科学原理一直没有得到科学阐明。本项目认为，"有故无殒"所强调机体状态对药物毒性应答的宏观差异性，在微观上看可能是由于机体在病理状态与正常生理状态的细胞微环境尤其是免疫微环境差异造成的。前期实验研究和文献调研结果初步证实了该假说的合理性。为此，本项目以含蒽醌中药（大黄、何首乌）肝毒性为研究载体和切入点，采用免疫学手段考查与大黄肝毒性"有故无殒"现象相关的细胞微环境关键因子，探讨"有故无殒"这一古老中医药毒性理论的现代生物学机制，也为含蒽醌中药肝毒性科学评价与合理用药减（避）毒提供基础科学依据。

《黄帝内经》“有故无殒”是最能反映中医辨证用药思想的理论认识之一，通俗地说即是“有病则病当之，无病则体受之”，对指导中药毒性评价与合理用药具有重要的理论和实际价值。然而，“有故无殒”的生物学机制和科学原理一直没有得到科学阐明。本项目首次从细胞微环境尤其是免疫微环境的角度，探讨研究了大黄对肝脏“有故无殒”现象的科学机制。.（1）实验证实了大黄、何首乌对肝脏的作用都存在“有故无殒”现象，其中何首乌的“有故无殒”现象是首次发现。即在特定剂量下大黄和何首乌对正常动物可引起肝损伤，但同样剂量对慢性肝病（肝纤维化）动物则体现出治疗作用。.（2）通过肝脏免疫微环境研究，结合特异性免疫细胞和因子的阻断实验反证，首次发现HMGB1  Kupffer Cell (M1/M2)  TNF-α、IL-1β、IL-6 / IL-10是大黄对肝脏“有故无殒”现象的生物学关键通路；其中在正常动物的损伤作用的关键通路为：HMGB1  M1 KC  TNF-α、IL-1β、IL-6；在肝纤维化动物的耐受作用关键通路为：HMGB1  M2 KC  IL-10；特别是分别阻断KC和HMGB1后，都可以显著降低正常动物对大黄肝损伤作用的应答程度，这与肝纤维化动物本身KC为抑炎状态的M2 KC并分泌抑炎因子IL-10所呈现的对大黄肝损伤的耐受作用是一致的。.（3）综合来看，肝脏免疫微环境的差异显著影响了肝脏对药物毒性损伤作用的应答程度，正常肝脏处在免疫可激活微环境，因此对药物的损伤作用较为敏感，易出现中毒损伤现象；而纤维化病理状态的肝脏处在免疫耐受微环境，因此对药物的损伤作用不敏感，不易出现中毒损伤现象，而同时药物的保护和治疗作用则可以显现出来，从而从宏观上表现为：“有病则病当之，无病则体受之”的“有故无殒”现象。.本项目首次从肝脏免疫微环境的角度，证实了“有故无殒”这一古老中医药毒性理论的科学合理性，并阐明了大黄对肝脏“有故无殒”现象的关键生物学通路机制，为含蒽醌中药大黄、何首乌肝损伤科学评价与合理用药减（避）毒提供基础科学依据。