ABSTRACT
Objective To investigate the effect of A53T α-synuclein on the expression of type 2 vesicular monoamine transporter (VMAT2) in neuronblastoma SH-SY5Y cells stably expressing A53T α-synuclein.Methods A53T α-synuclein eukaryotic plasmid was constructed by transfection of the SH-SY5Y cells using LipofectamineTM 2000, and a stable transfected monoclonal cell line was selected by G418.Western blotting and DCFH-DA staining were used to detect the effect of A53T α-synuclein overexpression on the expression of VMAT2 protein and level of reactive oxygen species (ROS).Results Western blotting showed that compared with the control group, the expression of VMAT2 protein was significantly decreased, and DCFH-DA staining showed that DCF signal was significantly increased (507.3 ±7.1) than that in the cell line stably expressing A53T α-synuclein (410.7 ±10.5) (P <0.05).Conclusions A53T α-synuclein can increase the intracellular ROS level by inhibiting the expression of VMAT2, thereby playing an important role in the pathogenesis of Parkinson′s disease.