Anthropogenic actions have direct and indirect impacts on natural systems, leading to significant alterations in marine ecosystems worldwide. One of the most notable problems is species loss, as the disappearance of species from an area can compromise ecological functions. This is at the core of a severe biodiversity crisis. To address and reverse these processes, marine protected areas (MPAs) have been utilized as a crucial tool to mitigate species loss, increase biomass, and serve as a fisheries management tool. However, there is a lack of information assessing MPAs from the perspective of their contribution to maintaining ecological functions. In recent decades, functional diversity (FD) indices have been widely used to assess ecosystem functioning. In this paper, we conducted an assessment using a global database of reef fish abundance to analyze the effect of No-Take Zones (NTZ) on the FD and “true” diversity (TD) indices of tropical reef fish assemblages in seven tropical biogeographic regions. We found a significant protective effect for some indices, although these responses were dependent on the bioregion. At the bioregional level, NTZs included lower numbers of species and functional entities than open access areas. Consequently, the functional richness protected within these zones partially represented the functional diversity in each biogeographic province. However, smaller-scale functional diversity indices responded to NTZ protection depending on the bioregion. Therefore, these results reinforce that the assessed NTZs are responsive to the protection of functional diversity, although they are not sufficient for safeguarding ecosystem functions in tropical reefs. This highlights the importance of expanding the number of protection entities worldwide with management strategies focused on coral reef fish functionality, as well as effective local/regional assessments. Thus, a new paradigm is necessary in the planning and creation of MPAs to safeguard ecosystem functions, with a priority given to the protection of ecosystem functions and habitats.