Objective To explore the effect of Moringa oleifera leaves, a characteristic plant resource in Yunnan Province, on the sleep improvement mechanism.Methods The chemical composition of ethanol extract from M. oleifera leaves (MOLEE) is determined by targeted metabolomics technology. The efficacy of MOLEE in improving sleep is evaluated in vivo through health food standard assays (mouse autonomous activity monitoring, pentobarbital sodium-induced mouse sleep test, and pentylenetetrazole-induced mouse convulsion test). Network pharmacology, molecular docking, and molecular dynamics simulation are employed in vitro to preliminarily investigate the mechanism of MOLEE on sleep-improving efficacy.Results MOLEE is rich in some active ingredients such as epicatechin, rutin, isoquercitrin, and apigenin, with their relative contents being 11.812, 18.071, 169.948, and 20.152 μg/mL, respectively. Compared with the blank group, the high-dose MOLEE group significantly reduces the number of autonomic activities of mice from 50.67 to 40.75 (P<0.05), and the sleep onset rate increases from 16.67% to 58.33%. The sleep latency of mice in the high- and low-dose groups of MOLEE are shortened to 76.14%, 76.39%, respectively (P<0.01). The sleep duration is prolonged from 24.35 to 46.77 and 46.54 min, respectively (P<0.01). Additionally, the high- and low-dose groups of MOLEE significantly reduce the mortality rate of pentylenetetrazole-induced convulsions by 41.67% and 33.33% (P<0.05), respectively. The results of network pharmacology indicate that there are 8 core components of MOLEE, namely epicatechin, rutin, isoquercitrin, apigenin, kaempferol, quercitrin, isoquercitrin, quercetin, corresponding to 399 targets. The Venn diagram shows that there are 95 common targets, and topological analysis shows that the 8 core components are closely related to key insomnia targets such as gamma-aminobutyric acid A (GABA-A), tumor necrosis factor (TNF), interleukin-6 (IL-6), and 5-hydroxytryptamine-A (5-HT-A) and other receptor proteins, and play a role in improving sleep. The results of GO and KEGG analysis indicate that the core components of MOLEE acted through regulating biological processes such as GABAergic neurotransmission, postsynaptic membrane potential regulation, and circadian rhythm, and through TNF, MAPK, 5-HT, and GABA signaling pathways. The molecular docking and dynamics simulations show that the eight core components are closely bound to the four key insomnia target proteins with strong stability, verifying the accuracy of the network pharmacology prediction results.Conclusion The material basis for MOLEE to improve sleep lies in its eight core components, including epicatechin and rutin, which exert their sleep-improving effects through multiple targets and multiple pathways.