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The present study was designed to investigate whether and how endothelin-1 (ET-1) impacted the generation of endogenous sulfur dioxide (SO<jats:sub>2<\/jats:sub>) in rat vascular smooth muscle cell (VSMC) proliferation and migration. Primary VSMCs and purified aspartate aminotransferase (AAT) protein were used in this study. We found that in the presence of ET-1, the expression of PCNA and Ki-67 was upregulated and the migration of VSMCs was promoted, while the AAT activity and SO<jats:sub>2<\/jats:sub>levels in VSMCs were reduced without any changes in AAT1 and AAT2 expression. SO<jats:sub>2<\/jats:sub>supplementation successfully prevented the ET-1-facilitated expression of PCNA and Ki-67 and the migration of VSMCs. Interestingly, ET-1 significantly increased reactive oxygen species (ROS) production in association with SO<jats:sub>2<\/jats:sub>\/AAT pathway downregulation in VSMCs compared with controls, while the ROS scavenger N-acetyl-L-cysteine (NAC) and the antioxidant glutathione (GSH) significantly abolished the ET-1-stimulated downregulation of the SO<jats:sub>2<\/jats:sub>\/AAT pathway. Moreover, the AAT activity was reduced in purified protein after the treatment for 2\u2009h. However, NAC and GSH blocked the hydrogen peroxide-induced AAT activity reduction. 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