Serum Pharmacology Combining Network Pharmacology to Discover the Mechanism of Total Flavonoids in <i>Aronia melanocarpa</i> Fruit for Acute Lung Injury

In this study, we used in vitro antioxidant assay and in vivo anti-inflammatory assay to comprehensively evaluate the activity of total flavonoids of Aronia melanocarpa fruit (AMF) in the treatment of acute lung injury (ALI) and explored the mechanism of action of total flavonoids of AMF in the treatment of ALI based on the method of serum medicinal chemistry combined with network pharmacology. In the present study, 107.37 mg g^-1 alcoholic extract of total flavonoids was used for the determination of in vitro antioxidant activity by UV spectrophotometric method; the ALI model mice were induced by lipopolysaccharide (LPS), and the organ index of mice, the wet-to-dry ratio of lungs, and the histopathological sections of lungs were measured; the levels of inflammatory factors (interleukin-1β [IL-1β], TL-6, and tumor necrosis factor-α [TNF-α]) were determined in serum and alveolar lavage fluid of mice by ELISA; the composition of AMF total flavonoids and the migratory components of blood were analyzed on the basis of UHPLC-MS/MS. We analyzed the composition of AMF total flavonoids and the migratory components in blood based on UHPLC-MS/MS technology as well as analyzed the active compounds, targets and mechanism of action of AMF total flavonoids in the treatment of ALI through serum medicinal chemistry combined with network pharmacology. The in vitro antioxidant combined with in vivo anti-inflammatory assays showed that AMF total flavonoids exhibited excellent therapeutic efficacy for the treatment of ALI. A total of 31 flavonoids were identified by UHPLC-MS/MS, and 21 compounds were identified from the serum samples of mice after the administration of AMF total flavonoids, including 11 prototypical compositions and 10 metabolites; 11 blood-migratory prototypical compositions and relevant targets for ALI were collected by network pharmacology. Finally, 77 intersecting targets were obtained, including AKT1, interleukin-6 [IL-6], TNF, EGFR, and IL-1β. These overlapping targets mainly corresponded to quercetin, 5-hydroxy-3,3',4',6,7,8-hexamethoxyflavone, and kaempferol, which regulate inflammatory responses through the PI3K/AKT signaling pathway. The mechanism of action of total flavonoids from AMF fruits for the treatment of ALI provided a new way to utilize AMF as a new food resource and new ideas for the research of new drugs for the prevention and treatment of ALI.