Objective To establish an anxiety-like gastric hypersensitivity (GHS) rat model of functional dyspepsia induced by new sequential stress. Methods Twenty-six male 1-day-old Sprague-Dawley rat pups were randomly divided into control and model groups (n=13 in each group). The model rats received sequential stress from postnatal day 2:neonatal maternal separation (NMS), acute gastric irritation (AGI) and restraint stress (RS). The control rats were reared freely with their mothers without undergoing the sequential stress. From postnatal 8^th week all rats started to receive elevated plus maze (EPM), open field test (OF), abdominal withdrawal reflex (AWR) and electromyographic test (EMG).Results EPM and OF experiments depicted that the model rats showed obvious anxiety-like behavior (P<0.05 or P<0.01). AWR and EMG tests exhibited that the model rats had elevated gastric hypersensitivity to gastric distention (P<0.05 or P<0.01).Conclusions An anxiety-like GHS rat model of functional dyspepsia can be successfully established with our new method of sequential stress.