The nurses often face moral challenges and conflicts as their duties are performed. In these situations, professional values and religious well-being support their performance within a specific framework and according to defined ethical principles. The purpose of this study is to examine the impact of ethics-based program on Iranian nurses' professional values and religious well-being. This was an experimental study conducted in Hajar and Ayatollah Kashani hospitals in Shahrekord (Iran) in 2019-2020. The participants included 130 nurses who were selected by convenience sampling and randomly assigned to the intervention and control groups. The outcome variables were professional values and religious well-being of nurses which measured at the beginning of the study, immediately, and two months after the intervention. Data collection occurred at baseline, immediately and 2 months after the intervention by using The Nurses Professional Values Scale-Revised (NPVS-R) and the Spiritual Well-Being Scale (SWBS). The study's findings revealed a statistically significant difference in mean changes scores related to professional values and religious well-being between the intervention and control groups following the implementation of the intervention. In conclusion, implementation of the ethics-based program can improve the professional values and religious well-being of nurses.