Ivermectin-Induced Apoptotic Cell Death in Human SH-SY5Y Cells Involves the Activation of Oxidative Stress and Mitochondrial Pathway and Akt/mTOR-Pathway-Mediated Autophagy

Abstract

Ivermectin (IVM) could cause potential neurotoxicity; however, the precise molecular mechanisms remain unclear. This study explores the cytotoxicity of IVM in human neuroblastoma (SH-SY5Y) cells and the underlying molecular mechanisms. The results show that IVM treatment (2.5–15 μM) for 24 h could induce dose-dependent cell death in SH-SY5Y cells. Compared to the control, IVM treatment significantly promoted the production of ROS, mitochondrial dysfunction, and cell apoptosis. IVM treatment also promoted mitophagy and autophagy, which were charactered by the decreased expression of phosphorylation (p)-Akt and p-mTOR proteins, increased expression of LC3II, Beclin1, ATG5, PINK, and Pakin1 proteins and autophagosome formation. N-acetylcysteine treatment significantly inhibited the IVM-induced production of ROS and cell death in SH-SY5Y cells. Autophagy inhibitor (e.g., 3-methyladenine) treatment significantly inhibited IVM-induced autophagy, oxidative stress, and cell apoptosis. Taken together, our results reveal that IVM could induce autophagy and apoptotic cell death in SH-SY5Y cells, which involved the production of ROS, activation of mitochondrial pathway, and inhibition of Akt/mTOR pathway. Autophagy inhibition improved IVM-induced oxidative stress and apoptotic cell death in SH-SY5Y cells. This current study provides new insights into understanding the molecular mechanism of IVM-induced neurotoxicity and facilitates the discovery of potential neuroprotective agents.

Chelerythrine-Induced Apoptotic Cell Death in HepG2 Cells Involves the Inhibition of Akt Pathway and the Activation of Oxidative Stress and Mitochondrial Apoptotic Pathway

“…To observe the nuclear morphological change, HepG2 cells were also stained with Hoechst 33342, according to our previous study [ 26 ]. In brief, HepG2 cells were treated with CHE at the various concentrations of 2.5, 5, and 10 μM for 24 h and were stained with Hoechst 33342 dye (at the final concentration of 10 μg/mL) for 30 min in the dark at room temperature.…”

“…The levels of caspase-9 and caspase-3 activities in HepG2 cells were examined using commercial caspase-9 and caspase-3 kits (Beyotime, Haimen, China), respectively. The detailed protocols were in line with our previous study [ 26 ]. In brief, HepG2 cells were treated with different doses of CHE (i.e., at 1.25, 2.5, 5, and 10 μM, respectively).…”

“…The levels of protein expression were examined using Western blotting and the detailed protocols were followed as per our previous descriptions [ 26 ]. In brief, HepG2 cells were treated with various doses of CHE (i.e., at 1.25, 2.5, 5, and 10 μM, respectively) or were treated with 0.2% DMSO as the vehicle control group.…”

Antioxidants, Oxidative Stress, and Non-Communicable Diseases

“…Ivermectin, a drug used in parasitic infections, has recently been linked to disinformation related to its efficacy against SARS-CoV-2. Zhang et al raised further awareness against self-medication and unsupervised use of ivermectin beyond the scope of its primary application, with an in vitro study linking this drug to neurotoxicity [ 12 ]. In this toxicological study, human neuroblastoma-derived SH-SY5Y cells were treated with increased doses of ivermectin, resulting in increased production of reactive oxygen species, mitochondrial dysfunction, and autophagy.…”

“…In this toxicological study, human neuroblastoma-derived SH-SY5Y cells were treated with increased doses of ivermectin, resulting in increased production of reactive oxygen species, mitochondrial dysfunction, and autophagy. The authors further identified the inhibition of the Akt–mTOR pathway as the cause of the apoptotic and autophagic events in this cell line [ 12 ].…”