Microplastics, recognized as some of the most pervasive and enduring pollutants, have emerged as a potential threat to environmental eco-health. While much is known about the effects of microplastics on soil microorganisms, our understanding of how they interact with terrestrial organisms and the underlying mechanisms remains limited. In this study, the effects of polyethylene microplastics at a concentration of 0.5 % (w/w) on the antioxidant enzymes, gut microbiota of Eisenia fetida and the soil microbiota on days 1, 3, 7, 15, and 30 were investigated. The results indicated that exposure to microplastics slightly increased the activities of superoxide dismutase (1.22-fold on day 3, 1.12-fold on day 7) and catalase (1.10-fold on day 3, 1.09-fold on day 7) in E. fetida, while exposure markedly decreased peroxidase activity (1.33- to 1.79-fold) throughout the whole period. Both the soil microbiota and the gut microbiota of E. fetida in terms of diversity and composition were significantly affected by the microplastic amendment, and their structure tended to be similar throughout the exposure time. The family Nocardiaceae was significantly enriched in both the soil and E. fetida gut biota with microplastic exposure. Our results demonstrated that the antioxidant enzyme response of E. fetida was closely related to both the microbiota, although this relationship with the gut microbiota may have been weakened by microplastic exposure. Overall, this study furnishes new perspectives on the ecotoxicity of microplastics, revealing significant implications for the vitality of soil-dwelling organisms and the overarching health of terrestrial ecosystems. Display omitted •LDPE microplastics altered the activity of antioxidant enzymes in Eisenia fetida.•LDPE microplastics altered the structure and composition of soil and gut microbiota.•Nocardia and Rhodococcus were enriched in both the soil and Eisenia fetida under exposure.•Correlation between antioxidase system and gut microbiota was weakened under exposure.