Neuropathic pain (NP) is a prevalent chronic condition frequently accompanied by adverse emotional states. Previous research has demonstrated the clinical efficacy of electroacupuncture (EA) in mitigating neuropathic pain and its associated mood disorders. Recent studies have underscored a correlation between gut microbiota and both NP and negative emotional states. Nevertheless, the relationship between the modulation of gut microbiota by EA and the amelioration of NP remains inadequately understood. Mice were randomly assigned to one of the three groups: the Control (Con) group, the EA group, and the Chronic Constrictive Injury (CCI) group (n = 12 each). Starting from the 8th day post-CCI induction, the EA group underwent EA treatment once every two days, for a total of 20 sessions. To investigate the impact of gut microbiota on CCI mice, we employed a variety of methods, including various behavioral tests and 16S ribosomal DNA (rDNA) sequencing. The results indicated that EA significantly ameliorated mechanical allodynia and emotional dysfunction induced by CCI in mice. Analysis through 16S rDNA sequencing revealed that the gut microbiota of NP model mice exhibited a marked increase in diversity. However, EA could partially reverse changes in the diversity of gut microbiota. The abundance of Alloprevotella, A2, Roseburia, Muribaculum, Ruminiclostridium, and Rikenella was increased, and the abundance levels of Bacteroides were decreased at the genus level in CCI mice. Following EA treatment, the relative abundance of Alistipes, A2, Roseburia, and Rikenella was decreased, whereas the relative abundance of Alloprevotella and Parabacteroides was increased in EA group when compared with the CCI group. These findings suggested that EA exerted a significant therapeutic effect on NP, potentially through modulation of the gut microbiota.