In this study, the impacts of gamma-irradiation from the low- and intermediate-level liquid radioactive wastewaters (LILW) to polyamide (PA) structures of nanofiltration (NF) membranes were investigated. As the gamma-irradiation increased to 300 kGy in the aqueous solution at 5 bar, both the salt rejection and the water permeability of NF membranes were decreased from 95.6 ± 0.1%–74.6 ± 0.5%, and from 33.7 ± 0.3 LMH to 21.4 ± 0.5 LMH, respectively. The surface free energy and Young's modulus of the membrane indicated the decrease in hydrophilicity and the increase in fragility of PA structure after gamma-irradiation. X-ray photoelectron spectroscopy and the streaming potential analysis exhibited that the gamma-irradiation resulted the increase in the cross-linked portion of the amide bonding from 28% to 45% due to the gamma-induced new bonding between unbound carboxylic groups and amine groups. Nuclear magnetic resonance analysis confirmed that the poly(p-phenylene) in polyamide structure were changed to poly(cyclohexane) and poly(cyclohexene) by hydrogen radical disproportionation generated from the gamma-irradiated water, and it is responsible to the increase of the cross-linked PA structures. The decrease in salt rejection and water permeability is attributed to the aging of PA structures by gamma-irradiation, thus, should be carefully monitored during the treatment of LILW using NF membrane processes.