iTRAQ proteomics analysis reveals that PI3K is highly associated with bupivacaine-induced neurotoxicity pathways

• Published in: Proteomics (Weinheim) Vol. 16; no. 4; pp. 564 - 575
• Main Authors: Zhao, Wei, Liu, Zhongjie, Yu, Xujiao, Lai, Luying, Li, Haobo, Liu, Zipeng, Li, Le, Jiang, Shan, Xia, Zhengyuan, Xu, Shi-yuan
• Format: Journal Article
• Language: English
• Published: Germany Blackwell Publishing Ltd 01.02.2016; Wiley Subscription Services, Inc
• Subjects: 1-Phosphatidylinositol 3-kinase; AKT protein; Anesthetics, Local - toxicity; Apoptosis; Apoptosis - drug effects; Autophagy; Autophagy - drug effects; BAX protein; Bioinformatics; Bupivacaine; Bupivacaine - toxicity; Caspase; Caspase-3; Cell Line, Tumor; Deoxyribonucleic acid; DNA; DNA damage; DNA Damage - drug effects; Extracellular signal-regulated kinase; FOXO1 protein; Humans; In Situ Nick-End Labeling; iTRAQ; JNK protein; Local anesthesia; Local anesthetic; Mitochondrial DNA; Neuroblasts; Neurons - cytology; Neurons - drug effects; Neurons - metabolism; Neurons - pathology; Neurotoxicity; Phagocytosis; Phosphatidylinositol 3-Kinases - metabolism; PI3K; Proteins; Proteomics; Reactive oxygen species; Reactive Oxygen Species - metabolism; Signal transduction; Signal Transduction - drug effects; Signaling; Target recognition; Local anesthetic; iTRAQ; Neurotoxicity; Bupivacaine; PI3K
• ISSN: 1615-9853; 1615-9861
• DOI: 10.1002/pmic.201500202

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.