Objective To examine the feasibility of using photothrombotic unilateral motor cortex lesion for establishing hindlimb spastic hemiplegia model in rats. Methods Twenty SD rats were randomized into 2 groups: rats in group A received erythrosine B injection followed by laser irradiation, and those in group B received sham operation. Then the rate dependent depression (RDD) of H reflex was employed to measure the spasticity of the plantaris before and 3, 7, 14, 28 days post-operation. H-E staining was used to observed the lesions in the motor cortex. Cholera toxin B subunit was used to retrogradely label motoneurons in the spinal cord. The number of vesicular glutamine transporter 1 (VGLUT1) boutons on motoneuron soma and dendrite was quantified by immunofluorescence staining and laser confocal microscope scanning. Results H reflex RDD in the right plantaris muscle was significantly decreased in group A compared with that in group B at 3, 7 ,14, and 28 days post-operation (P < 0.01). However, the H reflex RDD was not significantly different between groups in the left hindlimd at different time points after operation (P > 0.05). The number of VGLUT1 boutons on soma and dendrite in group A was significantly increased compared with that in group B (P < 0.01). H-E staining showed specific lesion in the motor cortex of the brain tissue in group A, but not in group B. Conclusion Unilateral photothrombotic motor cortex lesion is feasible to generate contralateral hindlimb spastic hemiplegia in rats.