Objective To investigate the therapeutic effect， underlying mechanism， and key genes involved in tetramethylpyrazine-pretreated umbilical cord mesenchymal stem cell （ucMSC） transplantation in a rat model of ischemic stroke. Methods The rat MCAO model was established， and umbilical cord-derived mesenchymal stem cells （ucMSCs） pretreated with or without tetramethylpyrazine were transplanted via the tail vein. Neurological function scores， TTC staining， and infarct rates were assessed. Localization of ucMSCs in brain tissue was observed. Experimental groups were analyzed using chip technology， and sample data were standardized. Bioinformatics analysis was employed to identify differential genes， which were subsequently validated by PCR. Results The treatment effect in ucMSCs pretreated with tetramethylpyrazine group was significantly superior to that of the untreated group， as evidenced by a significant reduction in neurological function score， infarct rate， and infarct area observed through TTC staining. Moreover， the treated group exhibited a significantly higher number of ucMSCs located within brain injury tissues compared to the untreated group. Subsequently， 2905 differential mRNA were screened based on predetermined criteria， including 1 754 up-regulated and 1 151 down-regulated genes. Among these differentially expressed genes related to the chemokine signaling pathway （identified using a multiple change value ≥ 2.0 and P value ≤ 0.05）， we identified 27 genes of interest. Notably， our analysis revealed activation of four genes closely associated with cell migration： Ccr6， Ccr3， Cxcr1 and Ccl6 respectively. Random verification experiments further confirmed a significant increase in gene expression for both Ccr3 and Cxcr1. Conclusions Pretreatment of umbilical cord-derived mesenchymal stem cells （ucMSCs） with tetramethylpyrazine significantly augmented the therapeutic efficacy in a rat model of ischemic stroke. Following pretreatment， there was a substantial increase in the migration of ucMSCs towards the site of brain injury. Our analysis suggests that this effect may be attributed to the activation of multiple chemokines， including Ccr6， Ccr3， Cxcr1， and Ccl6， by tetramethylpyrazine.