Abstract: The national land space is the carrier of carbon elements and is a vital site for carbon metabolism. The changes in the spatial pattern influence the regional carbon metabolism process. Optimizing this spatial pattern and maintaining the carbon metabolism balance are essential to achieve regional low-carbon, high-quality, and sustainable development. Using the Carbon Imbalance Index(CII) and the carbon conversion coefficient, this study analyzed the changes in the national land spatial pattern and spatial carbon emissions/sinks in Guizhou Province from 2000 to 2020. The ecological network analysis and ecological utility function were used to explore the carbon metabolism process and its ecological relationship induced by the changes in the national land spatial pattern. Appropriate suggestions were offered. The results showed that: (1) an increase in strong carbon emissions and a decrease in strong carbon sinks characterized Guizhou’s national land spatial pattern. Further, urban spatial carbon emissions had been decreasing since 2015, whereas agricultural spatial carbon emissions had been steadily increasing. The CII decreased from 47.5% in 2000 to 27.2% in 2020. (2) The positive, negative, and net carbon flows increased by 5.41, 15.70, and 28.76 times, respectively. Carbon metabolism processes remained negative, mainly due to the continued expansion of the urban space. (3) After 2015, the exploitation and control relationships showed a dominant (55%) upward trend, and the value of ecological utility function remained <1, indicating the negative impact of national land spatial pattern changes on carbon metabolism. This result was consistent with that shown by the net carbon flow. (4) The four ecological relationships exhibited considerable spatial heterogeneity. An increase from the central to the western region was indicated, while clustering characterized central and southwestern Guizhou during 2015-2020. This study quantitatively analyzed the changes in the carbon metabolism flow, metabolic anomalies, and the regional land spatial structure in Guizhou, and provided a scientific basis for regulating the regional carbon metabolism balance, towards a sustainable land spatial pattern.