Amiodarone is one of the most common medications for arrhythmia treatment, but it is generally considered as a secondary therapeutic option due to its severe side effects, including pulmonary toxicity. The direct toxicity of amiodarone in lung tissues have been discovered, including oxidative stress and apoptosis. Artemisinin is a clinical antimalarial drug, and its cytoprotective effect has been reported through inhibiting oxidative stress and apoptosis. The purpose of this study was to investigate whether amiodarone has direct toxic effect on human bronchial epithelial cells BEAS-2B and assess if artemisinin could protect BEAS-2B cells from amiodarone-induced cell injury and explore its underlying molecular mechanism. Our results of in vitro experiments showed that amiodarone could cause cell viability decrease, lactate dehydrogenase (LDH) release, reactive oxygen species (ROS) level increase, cysteine-aspartic acid protease (caspase) 3 activation and cell apoptosis in BEAS-2B cells. At the same time, artemisinin could upregulate the protein levels of calcium-/calmodulin-dependent kinase kinase 2 (CaMKK2), phosphorylated AMPK, nuclear factor erythroid 2-related factor 2 (Nrf2) and superoxide dismutase (SOD) 1 in BEAS-2B cells and protect cells from amiodarone-induced oxidative damage. The AMPK inhibitor Compound C could reverse the protective effect of artemisinin in BEAS-2B cells and primary human bronchial epithelial cells. In vivo experiments further proved that artemisinin could upregulate protein levels of phosphorylated AMPK and Nrf2 in lung tissues and protect the lung from amiodarone-induced apoptosis and bronchial epithelial cell shedding in mice. These results suggested that artemisinin can prevent airway epithelial cells and the lung tissue from amiodarone-induced oxidative stress and apoptosis through the activation of AMPK, it may provide new ideas for the prevention and treatment of amiodarone-induced pulmonary toxicity.