NCX3 alleviates ethanol-induced apoptosis of SK-N-SH cells via the elimination of intracellular calcium ions

• Published in: Toxicology in vitro 2021-04, Vol.72, p.105104-105104, Article 105104
• Main Authors: Xia, Zhixiu, Wang, Changliang, Wang, Xiaolong, Yu, Hao, Yao, Hui, Shen, Hui, Lan, Xinze, Wu, Xu, Zhang, Guohua
• Format: Article
• Language: English
• Subjects: Adenosine diphosphate; AKT protein; Apoptosis; Calcium; Calcium (intracellular); Calcium ions; Calpain; Caspase-3; Cell viability; Cyclic AMP response element-binding protein; Encephalopathy; Ethanol; Exposure; Flow cytometry; Fluo-3; Gene expression; Intracellular; Intracellular calcium; Memory tasks; mRNA; Na+/Ca2+ exchanger; NCX3; Nerve cell; Phosphorylation; Poly(ADP-ribose) polymerase; Polymerase chain reaction; Proteins; Reagents; Regulatory sequences; Ribose; Spatial analysis; Spatial memory; Time dependence; Western blotting
• ISSN: 0887-2333; 1879-3177
• DOI: 10.1016/j.tiv.2021.105104

Long-term alcohol intake may cause nerve cell apoptosis and induce various encephalopathies. Previously, we have shown that the expression of Na+/Ca2+ exchanger 3 (NCX3) was associated with the intracellular calcium concentration ([Ca2+]i) and apoptosis, involved in the spatial memory impairment in male C57BL/6 mice with chronic ethanol (EtOH) exposure. However, the mechanism involved is unclear. Here, we investigated the expression of NCX3 and its protective effect on SK-N-SH cells (a nerve cell line) after EtOH exposure. [Ca2+]i was measured using Fluo-3 AM reagent. Cell viability and the apoptotic rate were assayed using 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt (MTS) and flow cytometry, respectively. The expression of p-cAMP-responsive element binding protein1(p-CREB 1), NCX3 protein, and mRNA were observed using Western blotting and quantitative real-time polymerase chain reaction (qRT-PCR), respectively. Cleaved-caspase-3, caspase-3, rabbit anti- poly (ADP-ribose) polymerase-1 (PARP-1) and calpain-1 proteins were used to assess the degree of apoptosis. Our results showed that EtOH increased [Ca2+]i and apoptosis of SK-N-SH cells in a concentration- and time-dependent manner. The expression of NCX3 protein and mRNA was up regulated obviously after SK-N-SH cells were treated with EtOH. The phosphorylation levels of Akt and CREB 1 were up regulated in cells treated with EtOH. The expression of NCX3 protein was reduced in the SK-N-SH cells treated with Akt phosphorylation inhibitor (LY294002). The [Ca2+]i and apoptosis rate of SK-N-SH cells increased 1.31-fold and 1.52-fold after silencing NCX3 compared with those treated with 200 mM EtOH alone for 2 d. In contrast, the [Ca2+]i and apoptosis rate of SK-N-SH cells decreased 0.26-fold and 0.35-fold after overexpression of NCX3 in the 2 d-200 mM EtOH treatment group. These results suggest that NCX3 plays a critical role in neuronal protection via the elimination of intracellular Ca2+, which may be a promising target for the prevention and treatment of encephalopathy after ethanol exposure. •The expression levels of NCX3 in SK-N-SH cells can be regulated by ethanol exposure.•NCX3 can alleviate alcohol-induced apoptosis of SK-N-SH cells through eliminating [Ca2+]i.•Extracellular high Na+ can promot the NCX3's ability of Ca2+ clearance and counteract the toxicity of ethanol.