The effective medical applications of cold atmospheric pressure plasma jet (APPJ) have been reported by many researchers, including sterilization of liquid and solid surfaces, treatment of chronic wounds, treatment of cancer tumors, and blood clots. Therefore, in this study, we exposed Aporrectodea trapezoides and Eisenia fetida earthworms to APPJ discharge (0–30 s) to evaluate the impacts on regeneration of missed segments, apoptosis, lipid peroxidation (LPo), catalase activity (CAT), total antioxidant capacity (TAC), total proteins, and protein profile. Results showed APPJ induced significant effects on regeneration ability of earthworms after 20 and 30 s of exposure (p < 0.05). Atmospheric pressure plasma jet did not have significant effects on MDA content and TUNEL-positive cells, but this effect was significant for TAC and CAT in both species (p < 0.05). In conclusion, the present study revealed for the first time that regeneration of missed segments in earthworms can be stimulated by plasma treatment.