Exploring the Active Substances and Mechanism of <i>Physalis Calyx seu Fructus</i> Against Inflammation‐related Respiratory Diseases via Integrating Ultra‐Performance Liquid Chromatography‐Tandem Mass Spectrometry, Network Pharmacological Analysis, a

Physalis Calyx seu Fructus (PCF) is frequently prescribed for treating inflammation-related respiratory diseases in traditional Chinese medicine. The aim of the study is to identify the active partitions of PCF against inflammation-related respiratory diseases and explore their therapeutic effects and mechanisms. In the present study, two active partitions, 50-EFP-2 and 70-EFP, were obtained through anti-inflammatory bioactivity-guided enrichment of macroporous resin. These two partitions at 10 mg/kg potently alleviated lipopolysaccharide (LPS)-stimulated acute lung inflammation (ALI) and ovalbumin (OVA)-induced asthma in mice. Phytochemical investigation was performed using ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) and high-performance LC (HPLC), and indicated that withanolides are the predominant constituents of these two partitions. Moreover, their potential targets were predicted by network pharmacological analysis, which showed that withanolides in PCF mainly attenuated ALI and asthma by regulating nuclear factor-κB (NF-κB) to inhibit inflammatory response. 50-EFP-2 and 70-EFP inhibited inflammation and oxidative stress by regulating NF-κB and nuclear factor erythroid 2-related factor 2 (Nrf2), both in vivo and in vitro. Collectively, two partitions rich in withanolides were discovered from PCF, which potently alleviated ALI and asthma by regulating Nrf2 and NF-κB pathways. These findings provide the foundation for discovering withanolide-type leading compounds and developing novel therapeutics against inflammation-related respiratory diseases.