Objective To investigate the current status of nursing assistants’ attitudes and aging knowledge toward the elderly and to analyze their influencing factors. Methods Nursing assistants from West China Hospital of Sichuan University were selected by convenience sampling method between November and December 2021. A questionnaires were administered using the General Information Questionnaire, the Chinese version of the Attitude toward the Elderly Scale, and the Aging Knowledge Scale. Results A total of 391 nursing assistants were included. Nursing assistants’ attitudes toward older adults scored (126.26±19.79) and knowledge of aging scored (13.59±5.30). The results of multiple linear regression analysis showed that whether or not they had cared for older adults, whether or not they had received relevant training, whether or not they were willing to receive training, having worked as a caregiver for at least 5 years, and knowledge of aging were the main influencing factors on nursing assistants’ attitudes toward older adults (P<0.05). Conclusions The attitudes of nursing assistants toward the elderly in this group were on the positive level, yet their knowledge of aging requires improvement. Intrinsic learning motivation was identified as the strongest predictor of these attitudes. Therefore, systematic training should be designed to foster professional identity, integrate aging-related knowledge, and employ methods such as aging simulation experiences. This integrated approach can effectively promote the internalization of positive attitudes and enhance the quality of care.