Obesity is a state of accumulating excessive body fat, charactering by a high blood lipid and associating with various metabolic diseases. As a kind of dark tea, many studies revealed that long-term drinking Liupao tea (LT) can reduce weight (Liu et al., 2014). However, the anti-obesity mechanism and active ingredients of LT are not known.Liquid chromatography-mass spectrometry (LC-MS) combined with network pharmacology was used to screen the active components and related targets of Liupao tea water extract (LTWE). The key anti-obesity targets and pathways of LTWE were predicted by protein-protein interaction (PPI) networks, and enrichment analyses using Kyoto Encyclopedia of Genes and Genomes and Gene Ontology databases. Then, the active components selected by high-performance liquid chromatography (HPLC) fingerprinting were used together with LTWE in an adipogenic model and insulin resistance (IR) model in vitro.Most of the compounds identified from LTWE were flavonofids, esters, and amides. Key targets such as RAC-alpha serine/threonine-protein kinase, insulin, and tumor necrosis factor (TNF) were involved in the phosphatidylinositol-3-kinase-protein kinase B (PI3K-AKT) signaling pathway, pathways in cancer, and other pathways. Four active components were screened by network pharmacology combined with HPLC fingerprinting. The in vitro experiment of LTWE and its four active components showed that in insulin-resistant 3T3-L1 cells, LTWE, (-)-epigallocatechin gallate (EGCG) and gallic acid (GA) inhibited adipocyte differentiation. Three factors could inhibit the differentiation of 3T3-L1 cells by decreasing gene expression of peroxisome proliferators-activated receptor γ (PPARγ), fatty acid synthase (FAS), CCAAT/enhancer binding proteins-α (C/EBPα) and interleukin-6 (IL-6). Caffeine and ellagic acid (EA) showed opposite results, but their effects on promoting adipose differentiation diminished with increasing concentrations of drug. In dexamethasone-induced insulin-resistant 3T3-L1 cells, the fluorescence intensity of 2-Deoxy-2-[(7-nitro-2,1,3-Benzoxadiazol-4-yl)amino]-d-glucose revealed that LTWE, GA, EGCG, caffeine, and EA significantly promoted glucose consumption. LTWE, GA, and EA improved insulin resistance in adipocytes by upregulating gene expression of insulin receptor substrate-1 (IRS-1), PI3K, AKT, and glucose transporter 4 (GLUT4).LC-MS combined with network pharmacology preliminarianized that LTWE acts mainly on the PI3K-AKT signaling pathway. Cell experiments revealed that the anti-obesity effect of LTWE is the result of multi-component action, which inhibits the proliferation and differentiation of preadipocytes by regulating gene expression of adipogenic transcription factors and proinflammatory factors, and improves IR by activating the IRS-1/PI3K/AKT/GLUT4 pathway.Copyright © 2022 Elsevier GmbH. All rights reserved.