Objective To approach the associated mechanism by which α-synuclein (α-Syn) might regulate the metabolism of dopamine. Methods A DNA fragment, located at --495 to +25 of the human tyrosine hydroxylase (TH) gene, was amplified by PCR and inserted into the pGL3-Basic luciferase reporter vector. The recombinant plasmid pGL3-THprom was transfected into a dopammergic cell line MES23.5 or a α-Syn over-expressed MES23.5 (named MES23.5/hα-Syn^+). The promoter activity was detected by the Dual Luciferase Assay System. Results The luciferase activities in the MES23.5 cells transfected with pGl.,3-Basic, pGL3-THprom, and pGL3-Control vectors were 5.60±0.67, 26.80±4.11, and 32.90±4.75, respectively. On the other hand, the luciferase activity of pGL3-THprom in the MES23.5 (26.80±4.11) was significantly higher than that in the MES23.5/hα-Syn^+(14.40±0.61) (P〈0.01). Conclusion These results indicate that the -495 to +25 region in the TH gene possesses promoter activity for controlling the gene expression, and that α-Syn may negatively regulate the metabolism of dopamine by affecting the function of TH promoter as a trans-acting factor.