基于滋肾通关方PK-PD相关性研究阐释中医“反佐”配伍的作用机理

“corrigent” was a unique compatibility method in TCM prescription，however, its scientific connotation wasn’t clear. ZiShen TongGuan was the present prescription of “corrigent” compatibility theory. In view of the previous study found that the cinnamon which takes only 1/21 total amount of the prescription was necessary to maintain the curative effect of the whole prescription, We also found that cinnamon can increase the bioavailability of phellodendri’s effective component berberine, increase the absorption of berberine in kidney. It maybe a hint that cinnamon play the role of changing the pharmacokinetic of effective ingredients: phellodendron and anemarrhena and to increase the efficacy for the purpose of achieving "efficacy synergy". Accordingly, we hypothesised that " the compatibility of the mechanism cinnamon corrigent phellodendron and anemarrhena is its effective component change others’ effective ingredients in absorption, distribution, metabolism, distribution and its dynamics behavior when it transduced into cells ". To testify the aboved scientific hypothesis, we firstly adopt the pharmacological, pharmacokinetics and cell pharmacokinetics, and established the correlationship between the Pharmacokinetic and the Pharmacological effects. As to preliminary clarify the material basis of cinnamon " corrigent " phellodendron and anemarrhena, and its action mechanism we study the influence of cinnamon on pharmacokinetic behavior and efficacy of phellodendron and anemarrhena in normal and pathological condition by in vivo studies。

“反佐”是中医组方遣药的一种独特配伍方法,然而其科学内涵尚不明确。滋肾通关方是“反佐配伍”理论的代表，前期研究发现方中占总处方量仅1/21的肉桂是维持全方疗效所必需，且肉桂可以增加黄柏有效成分小檗碱的生物利用度及肾脏分布，提示肉桂可能具有改变黄柏、知母有效成分的药代动力学行为的作用，从而起到 “协同增效”的作用。据此，我们提出“肉桂反佐黄柏、知母的配伍机制是肉桂有效成分改变了黄柏和知母有效成分在体内的吸收、分布、代谢、排泄以及进入细胞的动力学行为”的科学假说。根据以上科学假说，项目拟在急性肾盂肾炎大鼠模型上，从药效学、药代动力学及细胞药代动力学等方面，基于PK-PD相关性研究，从体内、体外两方面研究肉桂对黄柏、知母有效成分药代动力学行为及药效的影响，以期初步阐明肉桂反佐黄柏、知母的物质基础，揭示中医反佐配伍的机制，从而指导临床的组方用药，提高方剂疗效。

“反佐”是一种中药配伍理论，“反佐”所用药味往往不超过2 味且药量仅占主药的1/6 左右，却可以在临床使用中产生奇效，其机制有待深入阐明。为了阐释其作用机制，本项目以南京中医药大学附属医院医院制剂滋肾通关胶囊为例，通过系统药理学的方法拟合“黄柏-知母”药对物质基础与作用靶点网络拓扑图，初步阐释了该药对的作用靶点与作用机制；建立了制备急性肾盂肾炎模型大鼠的方法、UPLC-MS/MS同时测定滋肾通关胶囊中活性成分的方法，并对滋肾通关胶囊及其拆方的活性物质在大鼠体内组织分布、组织与血浆样品的活性成分含量进行比较，及多剂量、单剂量药动学研究，同时采用代谢组学的方法对空白组、模型组、滋肾通关组、“黄柏-知母”组动物进行分析，得到差异代谢物，进行通路分析；以肾脏病理积分、血清IL-2、IL-α、IL-6、TNF-α等的检测为指标，系统评价滋肾通关方全方及拆方对急性肾盂肾炎的改善作用；同时建立LPS诱导的肾小管上皮细胞炎症模型，采用ELISA、实时定量PCR技术，评价滋肾通关方全方及拆方对肾小管上皮细胞（HK-2）分泌IL-1β，IL-6，TNF-α水平及mRNA的影响。本研究从中药活性物质在体内、体外的药动-药效学相互作用出发，同时采用代谢组学的分析方法多角度阐明肉桂“反佐”黄柏-知母的机制，对于深入诠释滋肾通关方的组方合理性及中医“反佐”理论的科学内涵具有重要的学术意义，可以为相关“反佐”中药方剂的配伍机制研究提供新的研究思路和方法，有助于更好地理解方剂的组方规律并推动其临床应用，进而提高方剂的临床疗效。本项目的实施过程中，发表论文6 篇，其中SCI论文2篇，硕士学位论文4 篇，申请专利1项。