title: iTRAQ proteomics analysis reveals that PI3K is highly associated with bupivacaine-induced neurotoxicity pathways.
abstract: Bupivacaine, a commonly used local anesthetic, has potential neurotoxicity through diverse signaling pathways. However, the key mechanism of bupivacaine-induced neurotoxicity remains unclear. Cultured human SH-SY5Y neuroblastoma cells were treated (bupivacaine) or untreated (control) with bupivacaine for 24 h. Compared to the control group, bupivacaine significantly increased cyto-inhibition, cellular reactive oxygen species, DNA damage, mitochondrial injury, apoptosis (increased TUNEL-positive cells, cleaved caspase 3, and Bcl-2/Bax), and activated autophagy (enhanced LC3II/LC3I ratio). To explore changes in protein expression and intercommunication among the pathways involved in bupivacaine-induced neurotoxicity, an 8-plex iTRAQ proteomic technique and bioinformatics analysis were performed. Compared to the control group, 241 differentially expressed proteins were identified, of which, 145 were up-regulated and 96 were down-regulated. Bioinformatics analysis of the cross-talk between the significant proteins with altered expression in bupivacaine-induced neurotoxicity indicated that phosphatidyl-3-kinase (PI3K) was the most frequently targeted protein in each of the interactions. We further confirmed these results by determining the downstream targets of the identified signaling pathways (PI3K, Akt, FoxO1, Erk, and JNK). In conclusion, our study demonstrated that PI3K may play a central role in contacting and regulating the signaling pathways that contribute to bupivacaine-induced neurotoxicity.
authors.affiliation: Department of Anesthesiology, Zhujiang Hospital, Southern Medical University, Guangdong Province, P. R. China.
authors.forename: Wei
authors.initials: W
authors.lastname: Zhao
authors.affiliation: Department of Anesthesiology, Zhujiang Hospital, Southern Medical University, Guangdong Province, P. R. China.
authors.forename: Zhongjie
authors.initials: Z
authors.lastname: Liu
authors.affiliation: Department of Anesthesiology, Zhujiang Hospital, Southern Medical University, Guangdong Province, P. R. China.
authors.forename: Xujiao
authors.initials: X
authors.lastname: Yu
authors.affiliation: Department of Anesthesiology, Zhujiang Hospital, Southern Medical University, Guangdong Province, P. R. China.
authors.forename: Luying
authors.initials: L
authors.lastname: Lai
authors.affiliation: Department of Anesthesiology, the Second Affiliated Hospital and Yuying Children's Hospital of Wenzhou Medical University, Wenzhou, Zhejiang, P. R. China.|Department of Anesthesiology, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pokfulam, Hong Kong, P. R. China.
authors.forename: Haobo
authors.initials: H
authors.lastname: Li
authors.affiliation: Department of Anesthesiology, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pokfulam, Hong Kong, P. R. China.
authors.forename: Zipeng
authors.initials: Z
authors.lastname: Liu
authors.affiliation: Department of Anesthesiology, Zhujiang Hospital, Southern Medical University, Guangdong Province, P. R. China.
authors.forename: Le
authors.initials: L
authors.lastname: Li
authors.affiliation: Department of Anesthesiology, Zhujiang Hospital, Southern Medical University, Guangdong Province, P. R. China.
authors.forename: Shan
authors.initials: S
authors.lastname: Jiang
authors.affiliation: Department of Anesthesiology, the Second Affiliated Hospital and Yuying Children's Hospital of Wenzhou Medical University, Wenzhou, Zhejiang, P. R. China.|Department of Anesthesiology, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pokfulam, Hong Kong, P. R. China.
authors.forename: Zhengyuan
authors.initials: Z
authors.lastname: Xia
authors.affiliation: Department of Anesthesiology, Zhujiang Hospital, Southern Medical University, Guangdong Province, P. R. China.
authors.forename: Shi-yuan
authors.initials: SY
authors.lastname: Xu
mesh_terms: D000779:Anesthetics, Local / Q000633:toxicity*; D017209:Apoptosis / Q000187:drug effects; D001343:Autophagy / Q000187:drug effects; D002045:Bupivacaine / Q000633:toxicity*; D045744:Cell Line, Tumor; D004249:DNA Damage / Q000187:drug effects; D006801:Humans; D020287:In Situ Nick-End Labeling; D009474:Neurons / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism*; D040901:Proteomics; D017382:Reactive Oxygen Species / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects*
chemical_list: D000779:Anesthetics, Local; D017382:Reactive Oxygen Species; D019869:Phosphatidylinositol 3-Kinases; D002045:Bupivacaine
keywords: Bupivacaine; Local anesthetic; Neurotoxicity; PI3K; iTRAQ