Zhisou powder displays therapeutic effect on chronic bronchitis through inhibiting PI3K/Akt/HIF-1α/VEGFA signaling pathway and reprograming metabolic pathway of arachidonic acid

Zhisou Powder (ZP), one of the most common prescriptions in traditional Chinese medicine, has been widely used in the treatment of acute or chronic bronchitis and chronic cough. The ZP was composed of Ziwan (Aster tataricus L. f.), Jiegeng (Platycodon grandiflorus (Jacq.) A. DC.), Jingjie (Nepeta cataria L.), Baibu (Stemona sessilifolia (Miq.) Miq.), Baiqian (Vincetoxicum glaucescens (Decne.) C. Y. Wu & D. Z. Li), Chenpi (Citrus × aurantium f. deliciosa (Ten.) M. Hiroe) and Gancao (Glycyrrhiza uralensis Fisch. ex DC.), with plant names among it checked with MPNS (http://mpns.kew.org). But until now, the key active components and targets of ZP, and related mechanism of ZP in the treatment of chronic bronchitis (CB) remain unclear.This study combined UPLC-Q-Exactive-Orbitrap-MS, network pharmacology, metabonomics with experiment verification to explore potential mechanism of ZP in the treatment of CB.UPLC-Q-Exactive-Orbitrap-MS was performed to analyze the chemical components of ZP. The potentially effective components, attractive targets and critical signaling pathways of Zhisou Powder in the treatment of CB were screened by UPLC-Q-Exactive-Orbitrap-MS combined with network pharmacology. Additionally, the CB model rats induced by SO2 were used to evaluate the anti-chronic bronchitis activity of ZP in vivo. The pulmonary pathology was determined by hematoxylin-eosin staining. Meanwhile, PI3K/Akt/HIF-1α/VEGFA signaling pathway predicted from network pharmacology was verified by Western blot and RT-PCR. Lastly, the metabolic changes of arachidonic acid (AA) in ZP-treated rats were quantitatively analyzed by LC-MS targeted metabonomics, and the proteins expression involved in AA metabolic pathway were detected by immunohistochemistry, immunofluorescence and Western blot.The main active components of ZP in the treatment of CB selected by network pharmacology and UPLC-Q-Exactive-Orbitrap-MS technology were quercetin, kaempferol, luteolin, galangin, isorhamnetin, naringenin, nobiletin, formononetin and so on. The core targets of these components were predicted to be TP53, TNF, IL-6, VEGFA, CASP3, IL-1β, JUN, PTGS2. Enrichment of KEGG pathway analysis found that PI3K/Akt/HIF-1α/VEGFA signaling pathway might play a key role in the treatment of CB with ZP. The in vivo study showed that ZP significantly improved the pathological changes of SO2-treated lung tissue and inhibited the activation of PI3K/Akt/HIF-1α/VEGFA signaling pathway. The changes of AA and its metabolites in vivo were studied by targeted metabonomics, and it showed that ZP could reprogram the disorder of AA metabolism which contributed to the treatment of CB with ZP.ZP displayed good therapeutic effect on CB model rats through inhibiting PI3K/Akt/HIF-1α/VEGFA signaling pathway to exhibit anti-inflammatory effect and reprogramming disordered metabolic pathway of arachidonic acid.