Ecotourism is an important part of the urban ecosystem. Understanding the impact of land use fragmentation in urban ecotourism areas on regional ecosystem service values can help promote the protection and rational development of urban ecotourism areas and maintain the stability of urban ecosystem services. We quantitatively assessed the value of terrestrial ecosystem services in the East Lake ecotourism area of Wuhan, from 2000 to 2015, using RS and GIS technologies. We determined the degree of land use fragmentation from a landscape pattern index and then clarified the temporal and spatial trends of land use and ecosystem service values. Finally, we analyzed the impact of fragmentation on ecosystem services. The results showed that (1) from 2000 to 2015, the area of land converted to construction in the East Lake ecotourism area was relatively large and the landscape showed obvious fragmentation trends: patch density increased from 1.67 to 1.96, and the landscape diversity index increased from 1.59 to 1.71. (2) During the 15-year study period, the value of overall services and function services of land ecosystems declined year by year. The total value decreased by 62.4881 million yuan, but the order of the value of various service functions remained stable; the top three functions were water conservation, waste treatment, and biodiversity conservation. Water-related services accounted for about 80% of the total service value. (3) There were significant positive correlations between the change in the area of grassland and cultivated land and the values of some regulatory and supply functions. Changes in the area of water had a significant negative effect on the overall value and the value of some supporting and cultural functions. There were significant correlations between changes in the patch density and the diversity indices of the landscape and the fragmentation index of forest lands and waters, and the change in values of some supportive and cultural functions. Fragmentation of the water area is a key factor in the decline of the overall value of ecosystem services and the value of cultural functions in the East Lake area.