The increasing climate change and human activities exert their influence on the ecological sensitivity of the region individually and interactively. Therefore, a clear understanding of the impact of climate change and human activities on ecological sensitivity will enhance the resilience of the regional ecological environment and the level of sustainable development. This study took the Yangtze River Economic Belt, the first demonstration zone of China's ecological civilization construction, as the research object. Based on the meteorological, remote sensing, and statistical data of 130 cities in the whole region from 2001 to 2021, an index system of climate change, human activities, and ecological sensitivity was constructed. Response surface methodology (RSM) was used to explore the effects of climate and anthropogenic single factors and interactions on the ecological sensitivity in each region. The results showed that: ① The ecological sensitivity value of the belt fluctuated and rose in time, rising by 2.2% from 2001 to 2021. In terms of space, the overall spatial distribution was "high in the north and low in the south." In 2021, the proportion of severely and extremely sensitive cities in the Yangtze River Economic Belt reached nearly 50%. ② For a single factor, the distribution of the effect of the same factor had certain characteristics: The areas where the single factors of economic development, rainfall, and temperature had a positive impact on the ecological sensitivity were concentrated in the areas with higher or faster economic development, along and south of the Yangtze River. For the interaction factors, the effect of 78.6% of the factors on the ecological sensitivity was negative interaction, and the change of one factor level would change the direction of the effect of the other factor on the regional sensitivity. ③ The comprehensive ecological management area of the Yangtze River Economic Belt was divided based on the ecological sensitivity and climate sensitivity. The governance areas that needed priority improvement were clustered within the three urban agglomerations and their northern adjacent areas, which meant that the ecological sensitivity and climate sensitivity of a city had spillover effects. This study is expected to provide inspiration for the economic zone and even the national and global efforts in the field of regional ecological governance.