This study investigated vitamin E's protective effects against spinosad-induced damage in adult male Wistar albino rats. Rats were administered vitamin E (200 mg/kg) and different spinosad doses (9 mg/kg and 37.38 mg/kg) orally. Intestinal tissues were collected for analysis on the first, third, and seventh days after administration. Lipid peroxidation (malondialdehyde [MDA]) and total glutathione (GSH) levels were quantified, and the structures of columnar epithelial cells in small intestine tissue were observed. Light, fluorescent, and electron microscope showed cell damage, such as deterioration in chromatin distribution and nuclear morphology, cell separation, large numbers of goblet cells, and impaired villus structure, in tissues collected from rats treated with spinosad compared to control rats. However, vitamin E ameliorated intestinal columnar cell damage. While GSH levels were lower in the 37.38 mg/kg spinosad group on all assay days, there was no significant difference in the 9 mg/kg spinosad group. The administration of vitamin E decreased in the 37.38 mg/kg spinosad group after the first day. The GSH levels in the 37.38 mg/kg spinosad with vitamin E group were lower than in the control group on the third and seventh days. However, while MDA levels were higher in the 37.38 mg/kg spinosad group on all assay days, there was no significant difference in the 9 mg/kg spinosad group. Nevertheless, MDA levels were higher in the 37.38 mg/kg Spinosad with vitamin E group compared to the control group on the third and seventh days. This study's findings highlight the need for further studies on spinosad and the protective role of antioxidants.