As a complex social ecosystem network, the area along the Grand Canal has a prominent contradiction between the demand for economic development and the protection of natural resources, which means that there is an urgent need for ecological restoration and environmental protection. Using ArcGIS, Conefor, Linkage Mapper and other software platforms, this paper developed an integrated analysis framework, through loose coupling of the attribute-function-structure index system and a series of methods such as the least cost path, circuit theory and moving window search. Based on the framework, we resolve a series of scientific issues in developing regional ecological networks, such as the selection of ecological sources, the simulation of potential ecological corridors, the assessment of the importance of ecological sources and corridors, and the identification of key ecological nodes. Moreover, an overall conservation pattern of the regional ecological network is constructed. The results show that: 1) A total of 88 important ecological sources are identified in the study area. The patches with high centrality values are mainly concentrated in the southern mountainous area and the areas with abundant rivers and lakes. 2) A total of 138 important ecological corridors are identified, and they are not evenly distributed. Extremely important corridors mostly appear between important patches, and very important corridors are mainly distributed in the central area. 3) Fifteen ecological pinch points are extracted, and they are mainly concentrated in the northern part of the study area and eastern Zhejiang Province. The barriers are mostly concentrated in the southern and northern parts of the study area. 4) Combining the demands of ecological protection and socioeconomic development, we propose an overall ecological conservation pattern of “one axis, five sections, multiple cores and multiple nodes” to effectively guide future ecological restoration work. These results can provide a useful reference and spatial guidance for decision makers in terms of ecological restoration and cooperation on cross-regional ecological protection along the Grand Canal.