锦灯笼干预慢性阻塞性肺疾病的物质基础与作用机制

Smoking and air pollution give rise to a high incidence of chronic obstructive pulmonary disease (COPD) in China. ‘Sweeping stagnate and sputum’ and ‘inhibitions of oxidative stress and inflammation’ are the therapeutic strategies for traditional Chinese medicines and western medicines, respectively. Calyx seu Fructus Physalis is a traditional Chinese medicine with the function of ‘sweeping stagnate and sputum’, and demonstrates a satisfying therapeutic effect on airway inflammation. Previous study indicated that Calyx seu Fructus Physalis possessed potent inhibitory effects against oxidative stress and inflammation, is consistent with therapeutic strategies of traditional Chinese medicines and modern medicines, and has a potential as new COPD therapeutic agent. It contained diverse bioactive substances. The mechanism on inhibition of oxidative stress and inflammation of these ingredients, with exception of flavonoids, are still unknown. Based on these findings, the work will be carried out as follows. (i) Bio-guided isolation and structure elucidation of ingredients with inhibitory effects against oxidative stress and inflammation, and illustrating the relationship between these two bioactivities and chemical ingredients. (ii) Based on the key oxidative stress- and inflammation- regulating signaling pathways, illustrating the molecular mechanism of diverse bioactive substances, and discussing the synergic mechanism of different structural skeleton. (iii) Evaluating the therapeutic effect of bioactive substances against COPD using cell and animal models, and discussing its druggability. The results of our research will illustrate its bioactive substances of Calyx seu Fructus Physalis, give the biological evidences for its traditional uses, provide a research basis for research and development of new COPD drugs, and have significance for the strategy of finding COPD therapeutic agents from traditional Chinese medicines.

吸烟和大气污染导致我国慢性阻塞性肺疾病(COPD)高发。“清痰瘀”与“抑制氧化应激和炎症反应”分别为中西医治疗COPD主要原则。锦灯笼清肺治咳、利咽化痰，治疗气道炎症疗效确凿。申请人前期研究发现，锦灯笼双重抑制氧化应激和炎症反应的作用显著，与中西医治疗COPD的原则相符，有成为干预COPD新药的潜力；其活性成分复杂、结构多样、种类繁多，除黄酮外，其他双重抑制氧化应激和炎症反应的活性成分尚未明确。本申请项目拟：（1）开展锦灯笼双重抑制氧化应激和炎症反应的活性成分追踪分离与构效关系分析；（2）针对氧化应激和炎症反应关键调控通路，揭示至少三种不同结构类型强活性成分的作用机制和多靶点协同作用；（3）在细胞和动物水平，开展活性成分干预COPD的有效性研究。研究结果将揭示锦灯笼药效物质基础，完善其中药理论和传统应用的生物学依据，为研发新型抗COPD药物奠定基础，对抗COPD中药的基础研究具有重要意义。

吸烟和大气污染导致我国慢性阻塞性肺疾病(COPD)高发，氧化应激和炎症反应是主要致病机理。锦灯笼具有清肺治咳、利咽化痰功效，中医临床治疗呼吸道炎症疗效显著。本项目以锦灯笼为研究对象，围绕Nrf2、NF-κB和细胞自噬三个氧化应激和炎症反应关键调控机制，针对慢性阻塞性肺疾病这一呼吸系统疾病，开展研究工作，包括： (1)开展了锦灯笼中氧化应激和炎症反应抑制活性成分追踪分离和结构确证，鉴定了84个化学成分（新化合物5个），包括甾体（主要为醉茄内酯）、黄酮、萜类等结构类型，评价了活性成分的氧化应激和炎症反应抑制活性，发现醉茄内酯和黄酮是主要活性成分类型。(2) 发现醉茄内酯类化合物DPB和黄酮类化合物NOR等通过促进Nrf2蛋白转位入核、增加Nrf2蛋白半衰期激活Nrf2调控的细胞防御机制，抑制砷、双氧水等氧化剂和毒物诱发的支气管上皮细胞和血管内皮细胞损伤。(3)建立了香烟烟雾诱发的小鼠急性肺部损伤模型和COPD模型并用于醉茄内酯4β-HWE和黄酮RN干预COPD有效性和作用机制研究，发现：4β-HWE能够上调Nrf2及其调控抗氧化基因的蛋白和mRNA水平，促进Nrf2蛋白与Keap1蛋白解离入核；4β-HWE可通过共价结合Keap1蛋白Cys151和Cys288半胱氨酸位点激活Nrf2抑制香烟烟雾诱发的小鼠COPD病理进展；黄酮RN能够通过与Keap1蛋白Cys151半胱氨酸共价结合激活Nrf2，并通过抑制NF-κB和诱导细胞自噬，缓解香烟烟雾诱发的小鼠肺部炎症反应。上述工作揭示了锦灯笼干预呼吸道炎症病变的药效物质基础，阐明锦灯笼传统用于治疗呼吸道炎症疾病的合理性和科学性，确证了活性成分成为COPD干预药物的潜力，为新型COPD药物研发奠定基础。