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BACKGROUND Artificial intelligence-driven chatbots are increasingly being used in health care, but few chat-based instant messaging support health education programs are designed for patients with chronic kidney disease (CKD) and evaluate its effectiveness and gender differences. Furthermore, there is little research documenting how CKD patients use the chat-based program that combining a chatbot with push notifications and feedback awards for improving patients’ communicative literacy and disease-specific knowledge. OBJECTIVE To assess the effectiveness and identify gender differences using a chat-based instant messaging support health education program among patients with chronic kidney disease and the usability of the program for CKD patients was also assessed. METHODS A pre-and post-study design was employed, and 60 patients were invited to join a three-month program for chronic kidney disease health education; 55 successfully completed the intervention. Data were collected from April to November 2020 using a structured questionnaire. Paired t-tests and generalized equation estimation were used to examine the intervention effectiveness and users’ evaluation. System Usability Scale provide the patients’ evaluation regarding the usability of the chat-based program. RESULTS Paired t-tests revealed significant differences before and after intervention on communicative literacy (t=3.99, p CONCLUSIONS The findings reveal that a chat-based instant messaging support health education program may be effective for middle-aged and older patients with CKD, showing similar intervention effectiveness between male and female patients, except for disease knowledge regarding prevention. The use of a chat-based program with multiple promoting approaches targeting middle-aged and older patients with CKD is promising, and users’ evaluation is satisfactory. CLINICALTRIAL The study was posted on www.clinicaltrials.gov (NCT05665517) on 23/12/2022.