The protection efficacy of nature reserves is a key element in achieving targets of biodiversity conservation. It is therefore very important to develop a scientific, systematic, and accurate index system for evaluating the protection efficacy of national nature reserves in China. Using methods of frequency statistics, expert consultation, analytic hierarchy process, and demonstration survey, we present a novel index system for evaluating the protection efficacy of Chinese national nature reserves for terrestrial vertebrates (excluding migratory birds) over a 10-year period. The indicator system included one target layer, two system layers, nine factor layers, and forty index layers. The system layer included ecological effectiveness evaluation (with a score of 60%) and management effectiveness evaluation (score of 40%). The ecological effectiveness evaluation was a comprehensive, dynamic evaluation of the target species, population, habitat, and ecological system. The management effectiveness evaluation was focused on the effectiveness of patrol and monitoring. The additional part aimed to analyze the impact of humans on the target species, population and nature resources of the nature reserve. This study combined the ecological effectiveness evaluation and the management effectiveness evaluation for the first time, highlighted the importance of time and space changes, distinguished the influence of natural factors from human factors, and integrated them into the evaluation results. By emphasizing quantifiable indicators, this evaluation index system could vastly assist the protection of nature reserves by improving management effectiveness, biodiversity conservation, and macroscopic decisionmaking.