细辛-干姜药对温肺化饮干预COPD-MH作用机理的代谢组学与转录组学整合研究

Chronic obstructive pulmonary disease (COPD) has the features of high morbidity, high disability and high mortality, which seriously endangers the lives and health of human beings. Airway mucus hypersecretion (MH) is a independent hazard affecting the progress and prognosis of COPD. COPD is induced by sputum in vivo and cold in vitro, and the pattern of TCM syndrome differentiation is cold fluid-retention suspend in lung. Asarum-dried ginger is one of the classic drug pairs of lung-warming decoction. Our previous studies predicted that the mechanism of action in the treatment of COPD was related to energy metabolism, oxidative stress and inflammatory reaction through network pharmacology. It may play a role through PI3K-Akt, VEGF, AMPK and other signaling pathways. Further studies on the treatment of COPD by asarum-dried ginger water decoction in rat models were conducted. Through high-throughput transcriptome sequencing analysis, it was found that the decoction indeed down-regulate the expression of PI3K mRNA, and is closely related to VEGF, AMPK and other signaling pathways, so as to antagonize the important biological marker MUC5AC of COPD-MH and exert its efficacy. Base on it, this project intends to use multi-histological integration study as a strategy to conduct correlation analysis of transcriptome-metabolomics data, fingerprint-metabolite spectrum data to verify the pharmacodynamic material basis and mechanism of asarum-dried ginger, thus to provide scientific basis for guiding clinical accurate and reasonable use of the drug pair.

慢性阻塞性肺病(COPD)具有高患病率、高致残率和高死亡率的特点，严重威胁人类的生命及健康。气道黏液高分泌(MH)是影响其病情变化及预后的独立危险因素。COPD因痰伏于内、寒诱于外，中医辨证为寒饮停肺证。细辛-干姜为温肺化饮的经典药对之一，课题组前期通过网络药理学预测其治疗COPD作用机制与能量代谢、氧化应激、炎症反应有关；可能通过PI3K-Akt 、VEGF、AMPK等信号通路发挥药效。进一步开展细辛-干姜水煎液治疗COPD大鼠模型动物实验，经高通量转录组测序分析，发现其确实能下调PI3K mRNA表达，并与VEGF、AMPK等信号通路密切相关，以此拮抗COPD-MH的重要生物学标志物MUC5AC而发挥药效。本项目拟在此基础上，以多组学整合研究为策略，将转录组学-代谢组学数据、指纹图谱-代谢物谱数据关联分析验证细辛-干姜药效物质基础与作用机制，为指导临床精准合理使用该药对提供科学依据。

慢性阻塞性肺病(COPD))具有高患病率、高致残率和高死亡率的特点，严重威胁人类的生命及健康，中医辨证为寒饮伏肺证。细辛-干姜（XGHP）为温肺化饮的经典药对之一，因此深入开展XGHP温肺化饮治疗COPD整体药效的分子机制，为指导临床精准合理使用XGHP提供科学依据。. 经过研究发现：XGHP有效化合物30个，主要包含细辛脂素、芝麻素、槲皮素等；XGHP组大鼠气管上皮黏膜损失少，腺体分泌物减少，炎症细胞浸润少；各组大鼠血清中的TG、TC、LDL-C、HDL-C、TNF-α、IL-6和MMP9水平表明XGHP可以通过调节COPD大鼠血脂水平，减少血液中促炎因子的水平来预防COPD的进展；XGHP能调控COPD大鼠核糖体、氧化磷酸化、FOXO信号通路、AMPK信号通路、嘌呤代谢、IL-17信号通路等；差异代谢物分析显示，XGHP组肌醇、甘露糖、正戊烷、亚牛磺酸水平显著降低，油酸、硬脂酸、棕榈酸、亚油酸水平显著升高，表明XGHP能够有效调节COPD大鼠肺脏中不饱和脂肪酸的代谢水平，涉及不饱和脂肪酸生物合成、亚油酸代谢、抗坏血酸和醛酸代谢、牛磺酸和亚牛磺酸代谢；整合转录组与代谢组分析显示FASN、SCD基因与油酸、棕榈酸、亚油酸代谢物之间存在联系，FASN、SCD和AMPK与XGHP的有效化合物对接良好；XGHP能够降低COPD大鼠肺组织的pAMPK蛋白表达，增加FASN、SCD蛋白表达。. 结论：（1）XGHP对COPD的肺组织具有保护作用，能够恢复大鼠血脂水平，降低血清中炎症指标，其作用的物质基础与L细辛脂素、芝麻素、芒柄花素、黄芩苷、紫铆素有关；这些单体将为后期进一步实验验证，深入阐述XGHP治疗COPD的具体物质基础提供依据和思路。. （2）XGHP可能通过AMPK信号通路，恢复FASN、SCD和pAMPK蛋白表达水平，从而增强不饱和酸的生物合成，恢复脂质代谢平衡，降低炎症反应。本研究还发现脂质代谢的紊乱能诱导炎症因子的释放，损伤血管内皮组织，是血管粥样硬化性改变的基础，加速血脂异常，由此可见，对COPD发病及治疗过程中，机体脂质代谢的研究是有意义的，为临床治疗提供了新思路。. （3）XGHP在机体内可能通过增加线粒体氧化磷酸化、脂肪酸氧化、非颤抖性产热治疗寒饮伏肺证。