Abstract: Objective: It was elucidated the anti-inflammatory active components and their mechanisms of action in the fruits of Diospyros lotus. Methods: Ultra-performance liquid chromatography-tandem quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS) was used to qualitatively analyze the chemical constituents of Diospyros lotus fruits, and combined with network pharmacology to explore the potential anti-inflammatory active components and their mechanisms of action, and validated the results by molecular docking. Results: A total of 26 flavonoid compounds, including 10 myricetin derivatives, 7 quercetin derivatives, 4 kaempferol derivatives, 2 apigenin derivatives, and 3 other flavonoid compounds, were identified from extract of D. lotus fruits based on molecular ion peaks and mass spectrometry cleavage characteristics in negative ion mode, among which 12 compounds were reported for the first time in the fruits. Network pharmacological analysis showed that compounds myricetin 3-sambubioside, myricetin 3-(2G-rhamnosylrutinoside), morin, myricetin 3-rhamnosyl-(1->2)-rhamnoside, isorhamnetin-3-O-glucoside, myricetin, quercetin, naringenin, apigenin and kaempferol had potential anti-inflammatory activities and exerted anti-inflammatory effects mainly related to PI3K-AKT, EGFR tyrosine kinase inhibitor resistance and cancer signaling pathway. Molecular docking showed that the active components stably docked with the key targets, and the results were consistent with the network pharmacological predictions. Conclusion: Flavonoids in D. lotus extracts have potential anti-inflammatory effects and are useful as adjunctive preventive agents in inflammatory diseases.