The establishment of nature reserves is a key approach for biodiversity conservation worldwide. However, there is a lack of unified methodology to evaluate the effectiveness of nature reserves, particularly in China, the world′s most populous nation supporting some of the most valuable biodiversity hotspots in the world. In this study, we conducted a long-term and large-scale analysis of the effectiveness of 20 of the earliest nature reserves established in Hainan Province, an island home to among the highest concentration of plants and animals in China. Remote sensing imagery from 1988, 1998, and 2008 were analyzed to investigate the temporal and spatial changes of natural forests in these nature reserves and surrounding areas. We also conducted transition matrix analysis and principle component analysis to identify the driving factors that affect the protection effectiveness of nature reserves. The results were as follows: 1) During the 20-year period from 1988 to 2008, natural forests coverage of the 20 studied nature reserves dropped 2.34 percentage points, whereas the natural forests coverage dropped 11.31 percentage points in a 0–5 km outside reserve buffer and 9.36 percentage points in a 5–10 km outside reserve buffer, indicating a significant inhibitory effect of the nature reserves on the loss of natural forests. 2) Natural forests coverage dropped in 60% of the studied nature reserves during the 20-year period, suggesting a poor protection effectiveness of these reserves, while the coverage proportion showed some increase(0%/yr–5%/yr) in other reserves. 3) Expansion of rubber and pulp forests as part of a booming economy were the main factors affecting the effectiveness of the nature reserves for conserving natural forests in Hainan Province. The results of this study provide an important empirical basis for the protection of natural forests in Hainan Province, which can be used as a blueprint for nature reserve evaluation in other places in China. The establishment of nature reserves is a key approach for biodiversity conservation worldwide. However, there is a lack of unified methodology to evaluate the effectiveness of nature reserves, particularly in China, the world’s most populous nation supporting some of the most valuable biodiversity hotspots in the study. we conducted a long-term and large-scale analysis of the effectiveness of 20 of the earliest nature reserves established in Hainan Province, an island home to among the highest concentration of plants and animals in China. sensing imagery from 1988, 1998, and 2008 were analyzed to investigate the temporal and spatial changes of natural forests in these nature reserves and surrounding areas. We also conducted transition matrix analysis and principle component analysis to identify the driving factors that affect the protection effectiveness of nature reserves. The results were as follows: 1) During the 20-year period from 1988 to 2008, natural for ests coverage of the 20 was decreased by 2.34 percentage points, while the natural forests coverage dropped 11.31 percentage points in a 0-5 km outside reserve buffer and 9.36 percentage points in a 5-10 km outside reserve buffer, indicating a significant inhibitory effect of the nature reserves on the loss of natural forests. 2) Natural forests coverage dropped in 60% of the studied nature reserves during the 20-year period, suggesting a poor protection effectiveness of these reserves, while the coverage proportion showed some increase (0 % / yr-5% / yr) in other reserves. 3) Expansion of rubber and pulp forests as part of a booming economy were the main factors affecting the effectiveness of the nature reserves for conserving natural forests in Hainan Province. The results of this study provide an important empirical basis for the protection of natural forests in Hainan Province, which can be used as a blueprint for nature reserve evaluation in other places in China.