Objective To investigate the neuroprotective effects of γ-secretase inhibitors N-［N-(3,5-difluorohenacetyl-L-alanyl)]-S-phenylglycinet-butylester(DAPT) on neural stem cells (NSCs) after cerebral ischemia in rats. Methods Thirty-three SD rats were randomly divided into three groups: model group, neural stem cell transplantation group (transplantation group) and DAPT+ transplantation group. In the other sham operation group, only the exposed blood vessels were separated under anesthesia. The neurobehavioral changes of rats in each group were observed 7 days after transplantation. The volume of cerebral infarction was observed by TTC staining. The pathological changes of brain tissue were observed by Nissl staining. The expression of BrdU/neuronal nuclei(NeuN) positive cells was detected by immunofluorescence double labeling. Results The neurological function score of the sham operation group was zero, no cerebral infarction, cell morphology was intact, Nissl bodies were abundant, and BrdU/NeuN positive cells were negative. Compared with the sham operation group, the neurological scores of the model group increased. There were obvious symptoms of neurological deficits, obvious cerebral infarction, disordered arrangement of nerve cells, nuclear pyknosis, rare Nissl bodies, and a significant decrease in the number of surviving neurons (P<0.05), with a small amount of BrdU/NeuN positive cells expression (P<0.05); Compared with the model group, the transplant group and the DAPT+ transplantation group had different degrees of neurological function score reduction, the cerebral infarction volume decreased, and the number of nerve cell survival increased (P<0.05), BrdU/NeuN expression of positive cells increased (P<0.05), and the indexes of DAPT+ transplantation group recovered most obviously. Conclusion DAPT can promote the differentiation of transplanted NSCs into neurons, and has neuroprotective effects on neural stem cells after cerebral ischemia.