TheGlycyrrhiza uralensis Fisch. is a common traditional Chinese medicine. However, its aerial part is currently not widely studied and used. Therefore, we aimed to investigate the neuroprotective effects of total flavonoids in aerial stems and leaves ofGlycyrrhiza uralensisFisch. (GSF) by anin vitroLPS-induced HT-22 cell model and anin vivo Caenorhabditis elegans(C. elegans) model. In this study, cell apoptosis was evaluated by CCK-8 and Hoechst 33258 staining in LPS-induced HT-22 cells. Meanwhile, ROS level, mitochondrial membrane potential (MMP), and Ca2+level were detected by the flow cytometer.In vivo,C. eleganswas also investigated the effect of GSF on lifespan, spawning, and paralysis. Moreover, the survival ofC. elegansto oxidative stimuli (juglone and H2O2), and the nuclear translocation of DAF-16 and SKN-1 were evaluated. The results showed that GSF could inhibit LPS-induced HT-22 cell apoptosis. Moreover, GSF decreased the levels of ROS, MMP, Ca2+, and malondialdehyde (MDA) and increased the activities of SOD and CAT in HT-22 cells. Furthermore, GSF did not affect the lifespan and laying of eggs ofC. elegansN2. However, it delayed paralysis inC. elegansCL4176 in a dose-dependent manner. Meanwhile, GSF increased the survival rate ofC. elegansCL2006 after juglone and H2O2treatment, increased SOD and CAT, and decreased MDA levels. Importantly, GSF promoted the nuclear translocation of DAF-16 and SKN-1 inC. elegansTG356 and LC333, respectively. Taken together, GSF can play a protective role in neuronal cells by inhibiting oxidative stress.