With the increasing aging population, chronic pain and musculoskeletal disorders have become significant health concerns, leading to a growing demand for non-invasive pain management strategies such as transcutaneous electrical nerve stimulation (TENS). While TENS is widely used for pain relief and neuromuscular regulation, gender differences in physiological responses remain underexplored. This study investigates gender-specific autonomic nervous system (ANS) and electromyography (EMG) responses to TENS stimulation at varying intensities (3VPP, 7VPP, and 11VPP) in 31 participants (16 males, 15 females). ECG and EMG signals were recorded using the BIOPAC MP36 system, and key HRV and EMG parameters were analyzed to assess neural and muscular activation patterns. Results showed that females exhibited broader neuromuscular responses at lower intensities, whereas males demonstrated increased autonomic activity at higher intensities, supporting the Gate Control Theory. These findings highlight the necessity of gender-specific electrostimulation therapy strategies and contribute to optimizing personalized TENS treatments.