title: The Ganglioside GM-1 Inhibits Bupivacaine-Induced Neurotoxicity in Mouse Neuroblastoma Neuro2a Cells.
abstract: OBJECTIVE
  Studies indicate that bupivacaine-induced neurotoxicity results from apoptosis. Gangliosides have been shown to promote neuronal repair and recovery of neurological function after spinal cord injury. Previously, we confirmed that in vivo administration of the ganglioside GM-1 attenuated bupivacaine-induced neurotoxicity in various animal models; however, the underlying mechanism remains unclear.
  
  
  METHODS
  Cells of the neuroblastoma line N2a (Neuro2a cells) were divided into three experimental groups: control, bupivacaine-treated, and bupivacaine-treated with GM-1 pretreatment. Cell viability and apoptosis were assessed through CCK-8 assays, Hoechst staining, and flow cytometry analysis of Annexin-V/propidium iodide double labeling. Real-time polymerase chain reaction and western blotting assessed the expression of caspase-3, caspase-8, and caspase-9.
  
  
  RESULTS
  Bupivacaine-induced apoptosis worsened with increasing dose and exposure time. Bupivacaine induced increased expression of caspase-3 and caspase-9, but not caspase-8, indicating that the mitochondrial pathway but not the death receptor apoptosis pathway was activated. GM-1 pretreatment inhibited bupivacaine-induced apoptosis and the expression of caspase-3 and caspase-9 in a dose-dependent manner.
  
  
  CONCLUSIONS
  Bupivacaine induced neurotoxicity by activating apoptosis via the mitochondrial pathway, and this was inhibited by GM-1 pretreatment.
authors: Department of Anesthesiology, The First Affiliated Hospital of Guangxi Medical University, 22 Shuangyong Road, Nanning, 530021, Guangxi, China.
authors: Yujie
authors: Y
authors: Liang
authors: Department of Anesthesiology, The First Affiliated Hospital of Guangxi Medical University, 22 Shuangyong Road, Nanning, 530021, Guangxi, China.
authors: Jiemei
authors: J
authors: Ji
authors: Department of Anesthesiology, The First Affiliated Hospital of Guangxi Medical University, 22 Shuangyong Road, Nanning, 530021, Guangxi, China.
authors: Yunan
authors: Y
authors: Lin
authors: Department of Anesthesiology, The First Affiliated Hospital of Guangxi Medical University, 22 Shuangyong Road, Nanning, 530021, Guangxi, China.
authors: Yajun
authors: Y
authors: He
authors: Department of Anesthesiology, The First Affiliated Hospital of Guangxi Medical University, 22 Shuangyong Road, Nanning, 530021, Guangxi, China.
authors: Jingchen
authors: J
authors: Liu
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D006690:Bisbenzimidazole / Q000378:metabolism; D015153:Blotting, Western; D002045:Bupivacaine / Q000633:toxicity*; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D005434:Flow Cytometry; D005677:G(M1) Ganglioside / Q000494:pharmacology*; D051379:Mice; D009447:Neuroblastoma / Q000473:pathology*; D000066829:Neuroprotection / Q000187:drug effects; D009498:Neurotoxins / Q000633:toxicity*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction
chemical_list: D009498:Neurotoxins; D012333:RNA, Messenger; D005677:G(M1) Ganglioside; D006690:Bisbenzimidazole; D002045:Bupivacaine
keywords: N2a cell; apoptosis; bupivacaine; gangliosides; neurotoxicity