Degeneration of cholinergic neurons is widely accepted as one of the main causes of cognitive impairments of Alzheimer’s disease (AD). However, studies on drugs that aim to improve the activity of the cholinergic neurons are limited, and there is currently no effective anti-AD drug. Therefore, the development of a new anti-AD drug remains to be looming. We investigated the neuroprotective effect of artemether on in vitro and in vivo models of AD. In vivo, artemether was used to treat scopolamine-induced AD model mice via intraperitoneal injection. Then, cognitive functional recovery, the activity of acetylcholine transferase (ChAT), acetylcholinesterase (AChE), synaptic plasticity, and the apoptosis of cholinergic neurons were assessed one month after the treatment. In vitro, HT22 cells were pre-incubated with artemether at different concentration for 2 hours and followed by were exposing to at 10 Aβ1-42 μM for another 24 hours. Subsequently, ROS levels, mitochondrial membrane potential, the activity of ChAT and AChE, neuronal cell apoptosis, and synaptic plasticity, as well as signaling pathway involvement in this process, were assessed using immunofluorescence and western blotting. Our results showed that scopolamine-induced AD mice treated with artemether had significant improvements in cognitive abilities, accompanied with increase in the activity of ChAT, reduction of AChE activity and apoptosis of neuronal cells. Artemether promoted the survival of cholinergic neurons, increased the expression of synaptic proteins, and alleviated the apoptosis of cholinergic neurons. Similarly, artemether significantly reversed the Aβ_1-42-induced increase of ROS levels, AChE activity, neuronal cell apoptosis, and synapse loss in vitro. This study shows that artemether has a neuroprotective effect on cholinergic neurons from injury and provides evidence for the clinical application of artemether.