利用免疫分析法解析黄连解毒汤中黄芩苷和栀子苷脑内时空分布特征

The compound pharmacokinetic characteristic in target site is one of the key links to reveal the material basis of traditional Chinese medicine. Huanglian Jiedu decoction was used as a model drug in this subject. The indirect competitive enzyme-linked immunosorbent assay (icELISA) methods based on baicalin and geniposide monoclonal antibodys are developed to detect the baicalin and geniposide concentration in the rat hippocampus, corpus striatum and striatum in normal physiological state and MCAO/reperfusion respectively. The aims of this subject are to explore the rules of baicalin and geniposide concentration in target sites changes according to the time, then compare the amount and the concentration distributed to the target site differences between MCAO/ reperfusion injury model and the normal physiological state rats, that is to analysis the correlation of baicalin and geniposide between dynamic process in brain and anti-cerebral ischemia effect, to reveal the role of baicalin and Geniposide in anti-cerebral ischemia effect. In addition, the dose-effect relationship can be demonstrated or explained by changes in the pharmacokinetic parameters especially in target site, so different dose of baicalin and Geniposide in Huanglian Jiedu decoction are researched to reveal the interaction relationship. Through this research we will partially reveal the material basis of Huanglian Jiedu Decoction in treating cerebral ischemia and provide direct evidence to explain its mechanism.

研究中药复方多成分在体内靶部位的药动学特征是阐明中药复方物质基础的关键环节之一。本课题以正常生理状态和MCAO/再灌注大鼠为模型，运用中药小分子单克隆抗体技术，建立黄芩苷和栀子苷的icELISA免疫分析方法，监测海马、纹状体等脑组织中黄连解毒汤两种主要成分浓度变化，探讨二者在靶部位浓度随时间变化的规律，比较分析MCAO/再灌注损伤模型大鼠与正常生理状态下分布到靶部位的量及药动特征的差异，即分析黄芩苷和栀子苷的协同动态过程与脑缺血保护药效的相关性。通过阐明黄连解毒汤中黄芩苷和栀子苷剂量变化造成的靶部位药物浓度变化，揭示多成分间的相互作用关系，为我们部分揭示黄连解毒汤产生药效的物质基础，为阐明其药效机理提供直接依据。

研究中药复方多成分在体内靶部位的药动学特征是阐明中药复方物质基础的关键环节之一。本课题比较了大鼠在正常生理状态和病理状态（MCAO/再灌注模型）下，注射给予栀子苷，经过不同时间后，主要指标成分栀子苷在动物体内的分布特征，栀子苷的血药浓度时间曲线均出现了双峰，MCAO模型组对栀子苷的吸收、消除速度和吸收量均大于生理组，两组之间没有显著性差异；栀子苷的脑组织分布结果显示，MCAO模型组栓塞侧大于未栓塞侧，统计具有极显著意义，MCAO模型组未栓塞侧与生理组没有显著差异。.按黄连解毒汤的原方中各主要成分（栀子苷、黄芩苷、盐酸巴马汀和盐酸小檗碱）的比例组成新的组分处方即组分组，与黄连解毒汤原方组进行对比实验，灌胃给予以上药物后，MCAO模型组结果显示原方组在海马、皮质和纹状体中的药物浓度与组分组的浓度相当；生理组与MCAO模型组相比，无论是血清还是海马、皮质和纹状体中，MACO造模后均使TNF-a的水平显著提高，给药后TNF-a的水平降低到与生理组水平相当，且组分组与原方组之间没有显著性差异，说明黄连解毒汤在抗中风后的炎症级联反应方面有确切的疗效，而且组分组与原方组的效果相当，说明组分组是其发挥药效作用的主要成分组，具有开发成现代组分中药的可能性。.另外，进行了黄连解毒汤中黄芩苷在小鼠体内的药代动力学研究及组织分布研究，建立了黄芩苷荧光免疫分析方法（FLISA），该方法更为简便、操作时间更短、灵敏度更高；进行了黄连解毒汤中黄芩苷在大鼠脑脊液中的药代动力学研究，为黄芩苷透过血脑屏障提供了直接依据。