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In recent years, the impact of global climate change and human activities on vegetation has become increasingly prominent. Understanding vegetation change and its response to climate variables and human activities are key tasks in predicting future environmental changes, climate changes and ecosystem evolution. This paper aims to explore the impact of Three Gorges Reservoir (TGR) water impoundment on the vegetation–climate response relationship in the Three Gorges Reservoir Region (TGRR) and its surrounding region. Firstly, based on the SPOT/VEGETATION NDVI and ERA5 reanalysis datasets, the correlation between climatic factors (temperature and precipitation) and NDVI was analyzed by using partial correlation coefficient method. Secondly, nonlinear fitting method was used to fit the mapping relationship between NDVI and climatic factors. Then, the residual analysis was conducted to evaluate the impact of TGR impoundment on vegetation–climate response relationship. Finally, sensitivity index (SI), sensitivity variation index (SVI) and difference index (DI) were defined to quantify the variation of vegetation–climate response relationship before and after water impoundment. The results show that water impoundment might have some impacts on the response of vegetation–climate, which gradually reduced with increasing distance from the channel; comparing with the residual analysis method, the SI and DI index methods are more intuitive, and combining these two methods may provide new ideas for the study of the impact of human activities on vegetation. [ABSTRACT FROM AUTHOR]