Based on network pharmacology and experimental validation,this study investigated the protective effect and mechanism of huperzine A (Hup A) on alveolar epithelial cells in acute lung injury (ALI).Firstly,the potential targets for Hup A to protect against ALI were identified by searching the public database for related targets of Hup A and ALI,and obtaining their intersection.Subsequently,the core target of Hup A protection against ALI was screened through construction of protein interaction network and "component-target-pathway" network,followed by verification through molecular docking.Furthermore,a cell model was established by stimulating mouse alveolar epithelial cells (MLE-12) with lipopolysaccharide (LPS) and pre-treating them with Hup A.The protective effect of Hup A was verified by detecting changes in cell viability,interleukin-6 (IL-6) level,super oxide dismutase (SOD) level and malondialdehyde (MDA) level.Finally,Western blot test was conducted to verify alterations in protein levels of key targets.Network pharmacological analysis revealed 144 potential therapeutic targets for Hup A,implicating its involvement in the regulation of biological processes such as protein kinase activity,oxidative stress,inflammatory response,and signaling pathways including phosphatidyl-inositol 3-kinase-serine-threonine kinase (PI3K-Akt) and mitogen-activated protein kinase (MAPK).Molecular docking results demonstrated favorable binding activity of Hup A with Akt1,Akt2 and Mapk1 (also known as extracellular regulated protein kinases,Erk).Cell experiments indicated that Hup A effectively ameliorated LPS-induced decline in cell viability (P< 0.05),reduced levels of IL-6,MDA and phosphorylated-Erk1/2/Erk1/2 (P< 0.05),while increasing SOD and phosphorylated-Akt1/Akt1 levels (P< 0.05).In conclusion,Hup A may exert a protective effect in LPS induced ALI through the regulation of Akt1 and Mapk1 targets within alveolar epithelial cells.