应用系统生物学结合PK-PD研究三芪口服液治疗膜性肾病的机制及物质基础

Idiopathic membranous nephropathy (IMN) is one of the most common causes (occupies about 30-40%) of primary nephrotic syndrome and its morbidity increased year by year. Due to its uncertain prognosis, it's quite difficult to decide the therapeutic schedule. Traditional Chinese medicine (TCM) is an effective method for the treatment of IMN and has received more and more attention. San Qi oral solution (SQ), which was designed and developed by Professor Yang Nizhi, is an effective formulated product to treat IMN and has been utilized for about 20 years in clinic. It is effective, and has been favored by many patients. Our study aims to elucidate the mechanisms of effects of SQ on treating IMN and its pharmacodynamic material foundation. Firstly, we would use molecular biology techniques combined with modern omics technology to study pharmacodynamics and the underlying mechanism. Secondly, we would compare the finger-print of SQ in vitro/vivo, and study the pharmacokinetics-pharmacodynamics (PK-PD) of SQ by using both animal and cell models, combined with correlation analysis of systems biology, thus illuminating the "dose-time-effect" of dynamic PK-PD in SQ treating IMN. Through this study, we try to comprehensively elucidate the mechanisms of effects of SQ on treating IMN and to provide theoretical support for clinical application.

特发性膜性肾病的发病率逐年增高，占原发性肾病综合症的30-40%，因特发性膜性肾病的预后在疾病发现之时无法进行准确判断，为治疗方案的选择带来困难，中医药是治疗膜性肾病的有效方法，越来越受到重视。三芪口服液是全国名老中医杨霓芝教授发明并应用于膜性肾病治疗的有效药物，临床应用近二十年获得患者好评。项目将针对三芪口服液治疗膜性肾病的机制及其药效物质基础展开研究，研究将分为两部分：首先，通过分子生物学技术与现代组学技术相结合，在分子-细胞-整体水平对三芪口服液治疗膜性肾病的药效机制进行研究；其次，通过构建三芪口服液体内/体外化学指纹图谱，结合细胞及动物模型探讨三芪口服液的PK-PD特点，结合系统生物学技术进行关联分析，阐明三芪口服液治疗膜性肾病的“量-时-效”的动态PK-PD过程，进而阐释三芪口服液治疗膜性肾病的药效物质基础。通过本研究以期全面阐释三芪口服液治疗膜性肾病的机制，为临床应用提供依据。

膜性肾病（MN）发病率逐年升高，是近年来唯一发病率增长的原发性肾小球疾病，给人们健康带来极大危害。三芪口服液（SQ）是临床治疗MN的有效药物，具有广阔的市场前景。本项目聚焦“SQ治疗MN”这一研究主题，应用“系统生物学联合药代动力学-药效学（PK-PD）”的特色研究方法开展药效机制及物质基础研究。研究内容分为四部分：第一，通过药效机制研究发现SQ能够通过NF-κB、Nrf2/NO-1两条信号通路保护足细胞，从而发挥减少蛋白尿、治疗MN的作用；第二，对MN患者临床样本的代谢组学及转录组学分析发现了影响MN进展的色氨酸、组氨酸及苯丙氨酸等氨基酸类代谢物及多种氨基酸代谢、不饱和脂肪酸代谢及糖酵解等代谢物变化途径，推测其可能与嘌呤代谢及肾素分泌等多条信号通路功能变化有关；第三，对SQ的化学表征及入血成分分析发现SQ主要包含人参皂苷、三七皂苷、芒柄花苷、毛蕊异黄酮等成分，并且SQ的主要单体成分均能入血，成为潜在药效物质基础。第四，基于上述研究结果，运用代谢组学联合PK-PD分析方法动态观察SQ干预后的MN模型大鼠体内药效成分及代谢物质变化，发现SQ主要化学成分在体内可通过影响氨基酸代谢治疗MN，其中谷氨酰胺和马尿酸可能是潜在靶点。本项目在整体药效及系统生物学分析基础上，应用代谢组学联合PK-PD技术从“整体-细胞-分子”多层水平及“结果-原因”双维角度，在“量-时-效”的动态变化中深入阐释了SQ治疗MN的药效机制及物质基础，丰富了SQ治疗MN的科学内涵，为中医药机制及药效物质基础分析提供了全新的研究思路，因其较高的外推性特征对同类型研究具有很好的借鉴意义。