Hygrophila salicifolia (Vahl) Nees is a traditional Chinese herbal medicine widely circulated in the Chinese medicinal materials market. It has gained significant popularity for its therapeutic effects in clearing heat and reducing swelling. The present study evaluated the chemical composition, antioxidant capacity, acetylcholinesterase, and α-glucosidase inhibitory effect of this species' essential oil. The major compounds of H. salicifolia essential oil were 1-octen-3-ol (29.0 %), phytol (5.7 %), phytone (4.6 %), cubebol (3.7 %), estragole (3.5 %), 7-epi-silphiperfol-5-ene (3.3 %), and ylangenol (3.1 %). The essential oil of H. salicifolia revealed antioxidant activity against both 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radicals with half-maximal inhibitory concentration (IC50) values of 2.13 ± 0.17 and 0.22 ± 0.02 mg/mL. The ferric reducing antioxidant power (FRAP) value was 170.97 ± 3.68 μmol·g−1. The essential oil has moderate activity against acetylcholinesterase (IC50 = 360.4 ± 12.7 μg/mL) and weak activity against α-glucosidase (14.2 % inhibition at 500 μg/mL). Moreover, we conducted in silico network pharmacology analysis and molecular docking to evidence the results of in vitro tests, providing a theoretical foundation for future drug development. The presented results suggest that the essential oil of H. salicifolia possesses the potential to be used in cosmetics, food, and pharmacological industries, emphasizing the need for further investigation.