To address the low charge capture capacity and poor wear resistance of triboelectric materials, lithium-ion intercalation is employed to synthesize monolayer–multilayer hybrid MoS2 nanosheets. Density functional theory calculations elucidate layer-dependent charge transfer and band structure modulation in the exfoliated MoS2, providing theoretical support for enhanced surface charge capture. By leveraging MoS2’s exceptional charge-trapping capability and tribological advantages, a MoS2/polytetrafluoroethylene (PTFE) composite film exhibiting high negative triboelectric polarity and ultralow friction characteristics is developed. The optimized 2.8 wt.% film exhibits a negative surface potential of −0.31 kV and a coefficient of friction (COF) of 0.07, representing a 416.7% increase in the absolute value of negative surface potential and a 65.5% reduction in COF compared to pure PTFE. The wind-driven triboelectric nanogenerator (TENG) delivers 452 V, 58 µA, and 6.98 mW at 19.5 m s−1, maintaining ≈95% of its initial open-circuit voltage over 21.6 million cycles. A self-powered heart rate monitor belt and respiratory sensor are fabricated, highlighting its potential for physiological monitoring. This work provides a strategy for triboelectric material design, offering a reference for developing high-performance and ultra-durable TENGs.