基于多模态生物影像技术检测炮制对黄连升降浮沉作用趋向的影响

Ascending, descending, floating and sinking (ShengJiangFuChen in Chinese, SJFC) is the core component of the property theories of Chinese herbal Medicine (CHM), the objective description of CHM effect tropism, and the important guiding principles for clinically accurate medication. As recorded, processing could change the SJFC tropism, with the general rules of "ascended by wine-processing, divergent by ginger-processing and descended by salt-processing". Traditional SJFC theory is always trapped by "analogy by classification" method, lacking objective quantitative criteria, and there is still no independent scientific evidence reporting nowadays. Multi-modality biological imaging method, including fluorescence molecular tomography (FMT) and infrared thermography (IRT), could qualitatively, quantitatively and image-based detect body physical and functional condition from different scale (body, organs, tissues, cells and molecular), so as to reflect the different of drug effect tropism and action site, which is confirmed with the concept of wholism in Traditional Chinese Medicine. Microdialysis (MD), an online and nondestructive sampling technology, is very suitable for the complex constituent metabolism analysis of CHM, could be applied for representation the SJFC effect tropism of CHM. Huanglian (Coptis chinensis Franch.), a common used heat-clearing and detoxifying herbs, is reported that processing could change its SJFC property, and its processed products, such as Jiu-Huanglian, Jiang-Huanglian and Yan-Huanglian could tend to Upper-Jiao, Middle-Jiao and Lower-Jiao respectively. In this research, Huanglian and its processed products are chose as research object, FMT, IRT and MD are combined for detecting the difference of biological imaging and chemical composition distribution of Huanglian effected on different model animal,so as to objectively evaluate the processing effecting on Huanglian ascending and descending tropism, and to provide scientific basis for the scientific nature confirmation of ascending, descending, floating and sinking property of Chinese herbal Medicine and the processing influence on them.

升降沉浮是中药药性理论的核心组成部分，是对中药作用趋向的客观描述,炮制可转变中药升降浮沉性能，并有"酒炙升提、姜炙发散、盐炙下行"的一般规律。然而，传统升降浮沉理论多囿于药类法象之缚，炮制对其影响也缺乏科学实证依据。那么，药性升降浮沉及炮制对其影响规律是否客观存在？炮制引起药物升降浮沉作用趋向改变的因素何在，是否与物质基础及分布趋向改变有关？机理又当如何？这些都是亟待深究的问题。多模态生物影像技术（FMT、IRT）可影像化地监测药物作用于机体的结构与功能趋向；微透析技术(MD)具有在体无损取样的特征，可研究中药复杂成分在体分布与作用趋向。本项目拟采用多模态生物影像技术联合微透析技术，以黄连及其炮制品（酒黄连、姜黄连、盐黄连）为研究对象，检测其作用于不同模型动物的影像学及化学成分分布差异，以期客观评价炮制对黄连升降浮沉作用趋向的影响，为证明中药升降浮沉及炮制对其影响的客观真实性提供科学依据。

升降浮沉是中药药性理论的核心内容之一，是指导临床用药准确性的重要依据，炮制可以改变中药升降浮沉药性，从而提高用药的针对性。本项目以黄连为例，制备了黄连不同炮制品（生黄连、盐黄连、酒黄连、姜黄连），复制了不同热证动物模型，模拟中医三焦热证，建立了微透析方法和黄连体内分布成分的LCMS测定方法，建立了黄连不同炮制品体内分布规律的PET测定方法，建立了黄连对不同细胞通道调控机制的luciferase测定方法。.采用Grace C18色谱柱（250 mm×4.6 mm,5μm），流动相为A：20mM乙酸铵-0.2%乙酸（pH3.8）,B乙腈，梯度：0min,5%B;10min,20%B;20min,25%B,40min, 40%B,流速0.6mL/min，DAD检测。MS条件为EMS+模式，检测分子量200-800。结果表明，酒制能提高黄连各炮制品生物碱类成分溶出，而盐制、姜制则降低各主要成分含量。建立的LCMS方法能够准确测定黄连不同炮制品在体微量化学成分的含量，客观评价炮制对黄连升降浮沉药性差异。.本研究采用luciferase assay，检测黄连不同炮制品对各细胞因子VDR、AR、ERα、ERβ、MR、TGFβ、NRF2、IRSE、UB、STAT3、AP1、TLR2、TLR4、GAS等细胞通路的调控作用，结果表明：黄连炮制品中，对多数细胞通道盐黄连（YHL）作用和生黄连（SHL）表现一致，而姜黄连（JHL）和酒黄连（JIUHL）作用一致且和前两者相反或加重，提示，盐制作用趋向沉降与生黄连固有作用趋向一致，而酒黄连和姜黄连的作用趋向为升浮，故而能缓解生黄连固有沉降趋势，且以酒黄连升提能力强于姜黄连，研究客观证实了炮制对黄连升降浮沉作用趋向影响的客观真实性。.项目同时探讨了黄连主要生物碱小檗碱、黄连碱、巴马汀对细胞通道调控作用的差异，结果表明，对多数细胞通道，小檗碱和巴马汀作用趋势一致，而黄连碱与二者作用趋势相反或加重，提示黄连存在不同作用趋向存在固有的物质基础，而炮制正是通过改变这种物质基础而发挥作用。.研究结果分别从化学成分、功能影像、分子机理的角度揭示了炮制对黄连升降浮沉药性影响的客观真实性，为阐释中药升降浮沉药性理论研究提供了可行路线和科学依据。