Objective In this study, we aimed to predict the inhibitory mechanism of shenlingcao oral liquid (SLC) on non-small cell lung cancer (NSCLC) by network pharmacology, and to verify it by molecular docking and in vivo animal experiments. Methods Active ingredients and corresponding targets of SLC and NSCLC were obtained by database and literature search. The common target of SLC and NSCLC was selected to construct the protein interaction network, and GO and KEGG enrichment analysis and molecular docking were performed. Lewis lung cancer mouse model was constructed, and Model, SLC high-dose group and SLC low-dose group, high-dose group and low-dose group were administered by intragastric administration at the dose of 8.75 g SLC lyophilized powder/kg and 3.50 g SLC lyophilized powder/kg, respectively. After 14 days of drug intervention, the tumor growth, pathological changes of tumor tissue and apoptosis of tumor tissue of tumor-bearing mice were observed, the changes of blood routine indexes of mice were detected, and the expression of p-AKT, AKT, p-PI3K, PI3K and Bcl-2 protein in tumor tissues of mice were detected, and the results of KEGG enrichment were verified. Results The network pharmacological analysis showed that there were 77 active ingredients, 618 potential targets, 1498 potential targets for NSCLC, 179 drug and disease intersection targets. Intersection target enrichment analysis showed that it was mainly concentrated in phosphatidylinositol 3 kinase signaling pathway (PI3K-AKTsignaling pathway), mitogen-activated protein kinase signaling pathway (MAPK signaling pathway) and other related pathways. Molecular docking showed that the top 10 core components had good bonding ability with the top 10 core targets. The results of animal experiments confirmed that compared with the Model, the tumor volume and weight in SLC high-dose and low-dose groups were significantly decreased (P < 0.05, 0.01), the expressions of white blood cells (WBC), neutrophils (Neu) and monocytes (Mon) were significantly decreased (P < 0.05, 0.01, 0.001), while the expressions of red blood cells (RBC), platelets (PLT) and hemoglobin (HGB) were significantly increased (P < 0.05, 0.01, 0.001), the apoptotic cells were significantly increased in early tumor tissue (P < 0.05, 0.01), and the protein expression levels of p-PI3K/PI3K, p-AKT/AKT and Bcl-2/GAPDH were significantly decreased (P < 0.05, 0.01). The expression levels of PI3K, AKT1 and Bcl-2 genes were significantly decreased (P < 0.05, 0.01, 0.001). Conclusion The mechanisms of SLC against NSCLC may be related to the activation of PI3K-AKT pathway and the promotion of apoptosis.