Abstract: This study conducted a quantitative evaluation of the relationship between biodiversity and human well-being in the Qianjiangyuan region, utilizing a robust indicator system of the contribution of biodiversity to human well-being. Based on multi-source data, including statistics, remote sensing and ground survey, this study evaluated the primary, secondary and tertiary indicators from the perspective of material supply, ecological regulation and spiritual culture, and revealed the spatial distribution differences and dynamic variations of biodiversity contribution to human well-being. The results show that, the spatial heterogeneity of human well-being index in the study area was strong, mainly attributed to the difference in natural resources. The highest human well-being index was observed in Kaihua County and Yushan County. Kaihua County displayed a higher contribution in ecological regulation, while Yushan County excelled in material supply and spiritual culture contributions. The spatial and temporal distribution of the contributions of material supply, ecological regulation and spiritual culture were significantly different. The material contribution exhibited a gradual increasing trend, with Yushan County being the highest and fastest growing in material supply. The ecological regulation contribution initially increased but later decreased. Kaihua County, Wuyuan County and Xiuning County had higher ecological contribution levels, benefiting from their natural advantages and larger forest ecosystem areas. The spiritual culture contribution index was greatly influenced by human subjective factors, with Yushan County standing out in the recreation index. Human well-being index has shown a trend of initial growth followed by a subsequent decline over the past 20 years. Yushan County has been growing with the fastest growth rate, which mainly manifested in the growth of material supply. The growth of material supply is the most significant indicator that contributed to the growth of human well-being in this evaluation system.