The construction of multi-function reservoirs is important to guaranteeing the development and utilization of water and hydropower resources, but the construction of any hydrologic projects will inevitably affect the downstream flow regime that provides the driving force for riverine ecosystems. This paper therefore aims to propose a framework for assessing multi-objective reservoir operation models based on flow regime using the middle and lower Yangtze River as a case study. Using indicators of hydrologic alteration (IHA) and the histogram matching approach (HMA), critical influential factors for flow were investigated at the Yichang and Datong gauges, which are typical of the middle and lower Yangtze River. This found the magnitude of annual extreme minimum water flow rates to be the most dramatically changed indicator, with an overall degree of hydrologic alteration at the Yichang and Datong gauges of 41.98% and 34.58%, respectively. The monthly mean discharges of February and October at Yichang have been significantly changed as a result of the Three Gorges Reservoir (TGR) operation. Meanwhile, an improved range of variability approach (IRVA) was developed to obtain ideal target ranges for monthly average flow. A multi-objective ecological reservoir operations model was created for the TGR; the Non-dominated Sorting Genetic Algorithm II (NSGA-II) was applied to the model. Three typical years of wet, normal and dry years were selected, and the operation results with corresponding ideal target ranges were analyzed for the downstream Yichang and Datong cross sections. The results showed that there should be a reduction by 23.57%, 28.10% and 39.92% in the monthly mean flow of February and an increase by 34.48%, 112.36% and 52.12% in the monthly mean flow of October at Yichang cross section in wet, normal and dry years, respectively.Close attention therefore needs to be given to integrating ecological objectives with appropriate flow regimes into multi-objective operation models.