Background and Purpose There has been increasing interest in the causes and pathogenesis of depression, which is a common psychiatric disorder. We aimed to investigate the protective effect of hops ethyl acetate extract (HEA) on neuroinflammation-mediated lipopolysaccharide (LPS)-induced depression-like symptoms. Experimental Approach A battery of behavior tests, including the open field test (OFT), elevated plus maze (EPM), tail suspension test (TST), and forced swimming test (FST), was used to evaluate the effects of HEA on LPS-induced depression. Furthermore, the levels of inflammatory factors (tumor necrosis factor-α [TNF-α], interleukin-1β) and norepinephrine were evaluated through enzyme-linked immunosorbent assay. The density of hippocampal dendritic spines was assessed through Golgi staining. Finally, the toxicological effects of hops extract on depression in mice were further analyzed through hematoxylin and eosin staining and blood biochemistry. Key Results Based on the OFT, EPM, TST, and FST results, oral gavage HEA prevented LPS-induced depression-like behaviors in the mice. Further, HEA reduced neuroinflammation, increased norepinephrine levels, and increased the density of hippocampal dendritic spines. Finally, blood biochemistry and HE staining did not reveal any side effects or toxicity of HEA. Conclusion and Implications Our findings indicated that HEA is a potential compound for treating depression.