Introduction: This work aimed to investigate the efficacy of electroacupuncture therapy on serum glycated hemoglobin (HbA1C) and plasma homocysteine (Hcy) in distal symmetric polyneuropathy (DSPN) in patients with type 2 diabetes mellitus (T2DM) of different genders. Methods: A retrospective analysis was conducted on clinical data of T2DM patients who visited our hospital for treatment from March 2021 to February 2024. A total of 70 patients (35 males and 35 females) were included. All patients received electroacupuncture treatment and were regularly followed up. Collected data included patients’ demographic information, disease characteristics, electroacupuncture treatment protocols, and treatment outcomes. Results: Following electroacupuncture treatment, clinical symptom scores, visual analog scale (VAS) scores, brief pain inventory scores, frequency of electroacupuncture treatments, and quality of life (QoL) scores in male patients were significantly lower to the female group. Plasma homocysteine (Hcy) levels in male patients were significantly lower than those in female patients. Moreover, 45.71% of male patients experienced a notable enhancement in symptoms, while 37.50% of female patients experienced notable symptom improvement. Additionally, male patients exhibited higher sensory nerve conduction velocity (SNCV) and the motor NCV (MNCV) relative to female patients. Conclusion: Electroacupuncture therapy has a certain efficacy in T2DM DSPN, but its efficacy varies among patients of different genders. This suggests the necessity of considering gender factors in formulating treatment plans to achieve individualized therapeutic strategies.