The natural abundance of nitrogen (N) stable isotopes (δ¹⁵N) has the potential to enhance our understanding of the ecosystem N cycle at large spatial scales. However, vegetation and soil δ¹⁵N patterns along climatic and edaphic gradients have not yet been fully understood, particularly for high-altitude ecosystems. Here we determined vegetation and soil δ¹⁵N in alpine grasslands on the Tibetan Plateau by conducting four consecutive regional surveys during 2001–2004, and then examined their relationships with both climatic and edaphic variables. Our results showed that both vegetation and soil N in Tibetan alpine grasslands were more ¹⁵N-enriched than global averages. Vegetation δ¹⁵N did not exhibit any significant trend along the temperature gradient, but decreased significantly with an increase in precipitation amount. In contrast, soil δ¹⁵N did not vary with either mean annual temperature or precipitation. Our results also indicated that soil δ¹⁵N exhibited a slight increase with clay content, but decreased with soil carbon:nitrogen ratio. A general linear model analysis revealed that variations in vegetation δ¹⁵N were dominantly determined by climatic variables, whereas soil δ¹⁵N was related to edaphic variables. These results provide clues for potential climatic and edaphic regulations on ecosystem N cycle in these high-altitude regions.