Bupivacaine Induces Reactive Oxygen Species Production via Activation of the AMP-Activated Protein Kinase-Dependent Pathway

Aims: It was our aim to investigate whether AMP-activated protein kinase (AMPK) mediates the considerable increase in reactive oxygen species (ROS) and cell apoptosis induced by bupivacaine in the human neuroblastoma cell line SH-SY5Y. Methods: The recombinant plasmids pGPU6/GFP/Neo-shRNA AMPK alpha 2 and pEGFP-N1-AMPK alpha 2 were constructed and transfected into the SH-SY5Y cell line. The expression of AMPK alpha 2 was determined by RT-PCR and Western blot after transfection. The SH-SY5Y cells transfected with recombinant plasmid were exposed to 1 mmol/l bupivacaine. Cell viability, intracellular ROS and apoptosis were determined. Results:The plasmid pEGFP-N1-AMPK alpha 2 can upregulate the expression of AMPK alpha 2, and the pGPU6/GFP/Neo-shRNA AMPK alpha 2 can downregulate the expression of AMPK alpha 2 in cells. Inhibition of AMPK alpha 2 expression attenuated ROS production and cell apoptosis, and overexpression of AMPK alpha 2 promoted ROS production and cell apoptosis after bupivacaine treatment. Conclusion: AMPK probably mediated ROS production and cell apoptosis induced by bupivacaine. Copyright (C) 2011 S. Karger AG, Basel