In vitro evaluation of the efficacy of azithromycin against the larvae of Echinococcus multilocularis

Alveolar echinococcosis (AE) is a serious zoonotic parasitic disease caused by the larvae of Echinococcus multilocularis. Given the limitations of current medications, there is an urgent necessity for the development of novel pharmacological treatments for AE. Azithromycin, a macrolide antibiotic, exhibits both antibacterial and antiparasitic effects. This study aimed to investigate its effects on the viability of E. multilocularis protoscoleces (EmPSCs) and to elucidate the underlying mechanisms. Ultrastructural changes were examined using scanning and transmission electron microscopy. Glucose assays, reactive oxygen species (ROS) assays, mitochondrial membrane potential (MMP) assays, and Western blot were used to evaluate the in vitro effects of azithromycin on EmPSCs. Our results demonstrated that azithromycin treatment significantly decreased the viability of EmPSCs and caused pronounced ultrastructural damage. Mechanistically, it significantly downregulated glucose transporter 1 (GLUT1) and protein kinase B (AKT) signaling, elevated ROS levels (P < 0.001), and decreased MMP. Furthermore, Azithromycin treatment significantly elevated the expression of the DNA damage marker γ-H2AX (P < 0.001), while also increasing lipid peroxidation and Fe²⁺ levels, depleting GSH content, and downregulating GPX4 and DHODH expression(P < 0.001). Collectively, azithromycin may exert multiple effects on EmPSCs in vitro, encompassing glucose metabolic disruption, oxidative stress, and ferroptosis.