Bupivacaine Reduces the Viability of SH-SY5Y Cells and Promotes Apoptosis by the Inhibition of Akt Signaling Pathway

Abstract

Bupivacaine (BUP) is a commonly used local anesthetic, while SH-SY5Y cells are a human neuroblastoma cell line frequently employed in research on neurotoxicity and neuroprotective mechanisms. To assess the neurotoxic effects of BUP on SH-SY5Y cells and the role of threonine-serine protein kinase B (Akt) signaling in BUP-induced nerve injury. SH-SY5Y cells were divided into three groups: the control group (Control), BUP group, and BUP + SC79 group. Cell viability was assessed using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, the level of reactive oxygen species (ROS) in cells was detected using the dihydroethidium fluorescence probe method, and changes in mitochondrial membrane potential were detected by flow cytometry, while BUP-induced apoptosis was evaluated by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining. The effects of BUP on Bax, Bcl-2, Caspase-3, Caspase-9, Akt and phosphorylated Akt (p-Akt) were analyzed by Western blot (WB). Compared with the control group, the BUP group and the BUP + SC79 group showed significantly reduced cell viability, significantly increased apoptosis, significantly elevated ROS levels, significantly decreased JC-1 polymer/monomer ratio, significantly increased protein levels of Bax, caspase-3, caspase-9, Akt, and p-Akt, and significantly decreased Bcl-2 protein levels (P < 0.05). However, compared with the BUP group, the BUP + SC79 group exhibited significantly increased cell viability (P = 0.022), significantly reduced apoptosis rate (P = 0.017), significantly decreased ROS levels (P = 0.015), significantly increased JC-1 polymer/monomer ratio (P = 0.024), significantly reduced protein levels of Bax, caspase-3, caspase-9, Akt, and p-Akt (P = 0.033, 0.028, 0.030, 0.035, and 0.005, respectively), and significantly increased Bcl-2 protein levels (P = 0.024). BUP can reduce the viability of SH-SY5Y cells and promote apoptosis, which may be related to its inhibitory effect on Akt protein activity.