The food industry has shown a significant interest in exploring plant-based components with lipid-lowering properties. Industrial hemp (Cannabis sativa L., IH), which is renowned for its rich bioactive constituents, has emerged as a potential botanical source. However, there are limited reports on the lipid-lowering effects of IH and its underlying mechanisms. Therefore, this study employed network pharmacology and a high sucrose (HS)-fed Drosophila melanogaster larvae model to investigate the lipid-lowering effects of IH. Network pharmacology analysis confirmed that flavonoids, sterols, flavonoid glycosides, and phenolic compounds present in IH possess lipid-lowering effects, suggesting that IH may exhibit potential lipid-lowering activity. Subsequently, in vivo validation demonstrated that IH significantly reduced triglyceride levels in a concentration-dependent manner in the HS group. Notably, a 1 % concentration of IH markedly decreased lipid accumulation and was selected for further investigation of its mechanism in HS-fed larvae. Transcriptome sequencing analysis revealed that the lipid-lowering effect of IH is associated with the regulation of lipid metabolism and related signaling pathways, including the MAPK signaling pathway, HIF-1 signaling pathway, AMPK signaling pathway, cAMP signaling pathway, and cGMP-PKG signaling pathway. These results are consistent with the findings from the network pharmacological analysis and confirm the role of IH in modulating lipid metabolism. In conclusion, this study demonstrates that IH has significant lipid-lowering characteristics and holds great potential for future functional food applications. (c) 2025 SAAB. Published by Elsevier B.V. All rights are reserved, including those for text and data mining, AI training, and similar technologies.