Ecosystem service flow plays a vital role in the formation, transportation, transformation, and maintenance of ecosystem services. For ecosystem services with spatiotemporal mismatch of supply and demand, ecosystem service flow explains the integrated process of ‘supply-flow-demand’ of ecosystem services. The present study evaluates the supply and demand of ecosystem water provision services in the Qinghai-Tibet Plateau and simulates the spatial flow pattern and transmission mechanism of water provision services. Additionally, the study establishes dynamic and static water security indices and identify water security level to quantify the water resources security of Qinghai-Tibet Plateau under the changing ecological environment. The research shows: (a) the annual total water surplus were 6.71 × 1011 m3, 8.43 × 1011 m3, 7.86 × 1011 m3 and 2.91 × 1011 m3. The supply-demand relationship of water provisioning service in the Qinghai-Tibet Plateau shows an obvious oversupply condition; (b) The water security level of the plateau is high (level V and level IV), indicating that the water security status of the Qinghai- Tibet Plateau is under good condition, however, the area with low-security levels (Level I and Level II) is increasing. (c) From the perspective of ‘supply-demand-flow’ of ecosystem services, although the function of the water tower on the Qinghai Tibet Plateau has declined, it remained safe condition on the whole study area. The method of establishing spatial correlation between mismatched supply and demand of ecosystem services and evaluating regional water security based on ecosystem service flow constructed in this study presents the water security status and spatial distribution of Qinghai-Tibet Plateau more scientifically, providing a reference for water resource management in other regions. © 2022 The Author(s). Published by IOP Publishing Ltd.