基于尿液和胆汁中化学信息逆向追踪研究锦灯笼抗炎药效物质基础及其作用机制

Physalis alkekengi L. var. franchetii has the effects of clearing away heat and toxic material and relieving sorethroat and dissipating phlegm. It is found in our previous research that the extract of Physalis alkekengi L. var. franchetii had significant anti-inflammatory effect, and the components of physalins,flavonoids and megastigmane glycosides from this plant showed notable anti-inflammatory activity in vitro. However,these constituents could not be detected in vivo after being given normal medicinal dose of the extract of this plant. Consequently, how to identify the effective components becomes the difficulty of research of Physalis alkekengi L. var. franchetii. We put forward the assumption that metabolites might be the effective components in vivo based on our previous work. We plan to clarify the chemical information from the urine and bile of rats after intragastric administration of the extract of Physalis alkekengi L. var. franchetii, and track conversely the in vivo existence form of the extract constituents, accompanied by examination of the anti-inflammatory activity of these in vivo components and their possible mechanism. And then, we will investigate the relationship between pharmacokinetics and pharmacodynamics, and elucidate the anti-inflammatory substances. The research plan sufficiently focuses on how the traditional Chinese medicine shows its effectiveness in vivo.This research will not only explain actual effective substances of Physalis alkekengi L. var. franchetii, but also reveal the its effect mechanism.This project will provide enlightenment for the research of effective substance of traditional Chinese medicine and establish a new way for the modern research of traditional Chinese medicine.

锦灯笼具有清热解毒、利咽化痰之功效。前期研究发现锦灯笼提取物具有确切的抗炎作用，其含有的酸浆苦素类、黄酮类及单萜苷类体外表现出显著的抗炎活性,但锦灯笼正常用药剂量下这些成分在体内检测不到,如何确定锦灯笼的体内实际药效成分成为锦灯笼深入研究的瓶颈。申请人在前期工作基础上提出了"代谢产物可能是锦灯笼体内发挥药效的主体"的假设，拟从大鼠灌胃锦灯笼提取物易于富集的尿液和胆汁出发，寻找与锦灯笼相关的化学物质信息，以此为导向逆向追踪体内及提取物中的存在形式，并通过抗炎生物活性评价明确体内药效形式及其可能机制，进而以体内活性形式为指标探讨药物动力学与药效动力学之间的相关性，最终明确锦灯笼的体内外抗炎药效物质。课题的研究思路充分考虑到口服中药的作用特点，研究工作不仅可阐明锦灯笼药材的真实药效物质，还可揭示其体内的作用机制，对传统中药药效物质研究具有重要的启示作用，同时为传统中药的现代研究开辟一条新的途径。

本课题以锦灯笼为研究对象，从大鼠灌胃锦灯笼提取物后易于富集的尿液和胆汁出发，逆向追踪研究锦灯笼的抗炎药效物质基础。在大鼠灌胃给予锦灯笼提取物后的尿液和胆汁样品中共鉴定出16个原形成分和51个代谢产物。进一步研究发现锦灯笼提取物灌胃后吸收入血成分有33种, 其中18种为代谢产物。主要成分Luteolin-7-O-glucopyranoside和physalin A的体内代谢研究结果表明，水解、还原、葡萄糖醛酸结合、硫酸结合、甲基化、磺化和羟基化代谢是锦灯笼化学物质在大鼠体内的主要代谢途径。大鼠灌胃给予锦灯笼提取物后的药代动力学初步结果证明，在所给药剂量下主要黄酮类化合物的原形形式在血浆中几乎检测不到；physalin A、physalin D和physalin L吸收较快且于1.3小时左右达到峰浓度。体内抗炎作用评价明确了黄酮、酸浆苦素能显著抑制二甲苯诱导的小鼠耳肿胀，是锦灯笼发挥药效的主要物质基础。锦灯笼的抗炎作用机制可能为清除DPPH自由基，抑制LPS诱导的巨噬细胞释放炎症介质NO，且黄酮类和酸浆苦素类化合物的抗炎功效存在协同作用。本课题建立了一套基于排泄物化学信息的中药药效物质研究新方法，并应用该方法初步阐明了锦灯笼的药效物质，揭示了其体内过程和作用机制，为传统中药的现代研究开辟一条新的途径。