title: Bupivacaine induces reactive oxygen species production via activation of the AMP-activated protein kinase-dependent pathway.
abstract: OBJECTIVE
  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 AMPKα2 and pEGFP-N1-AMPKα2 were constructed and transfected into the SH-SY5Y cell line. The expression of AMPKα2 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-AMPKα2 can upregulate the expression of AMPKα2, and the pGPU6/GFP/Neo-shRNA AMPKα2 can downregulate the expression of AMPKα2 in cells. Inhibition of AMPKα2 expression attenuated ROS production and cell apoptosis, and overexpression of AMPKα2 promoted ROS production and cell apoptosis after bupivacaine treatment.
  
  
  CONCLUSIONS
  AMPK probably mediated ROS production and cell apoptosis induced by bupivacaine.
authors.affiliation: Department of Anesthesiology, Zhujiang Hospital, Southern Medical University, Guangzhou, China.
authors.forename: Jun
authors.initials: J
authors.lastname: Lu
authors.forename: Shi Yuan
authors.initials: SY
authors.lastname: Xu
authors.forename: Qing Guo
authors.initials: QG
authors.lastname: Zhang
authors.forename: Hong Yi
authors.initials: HY
authors.lastname: Lei
mesh_terms: D055372:AMP-Activated Protein Kinases / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D000779:Anesthetics, Local / Q000009:adverse effects*; D017209:Apoptosis / Q000187:drug effects; D015153:Blotting, Western; D002045:Bupivacaine / Q000009:adverse effects*; D018929:Cell Culture Techniques; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D005434:Flow Cytometry; D049452:Green Fluorescent Proteins / Q000235:genetics; D006801:Humans; D010957:Plasmids; D034741:RNA, Small Interfering / Q000235:genetics; D017382:Reactive Oxygen Species / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection
chemical_list: D000779:Anesthetics, Local; D034741:RNA, Small Interfering; D017382:Reactive Oxygen Species; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; C526181:PRKAA2 protein, human; D055372:AMP-Activated Protein Kinases; D002045:Bupivacaine