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. 2020 Dec 3;14:583782. doi: 10.3389/fncel.2020.583782

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Copyright © 2020 Piao, Wang, Liu, Wang, Chen, Qin, Ge and Feng.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Sevoflurane induced DNA damage in neuronal cells. (A) Images of neuronal cells acquired from fluorescence microscopy after alkaline comet assay. SH-SY5Y cells and HT22 cells treated with 4 and 8% sevoflurane for 12 h showed more cells with comet tails than that in control group, and the higher concentration, the longer the comet tails. Pretreatment of neuronal cells with antioxidant NAC at 5 mmol/L markedly prevented sevoflurane-induced appearance of comet tails in SH-SY5Y cells and HT22 cells. (B,C) Statistical analysis of alkaline comet assay revealed that sevoflurane induced a significant increase in the percentage of cells with comet tails and DNA content within the tails (p < 0.01), which were markedly inhibited in the presence of NAC (p < 0.01). (D) Images of neuronal cells acquired from fluorescence microscopy after neutral comet assay. Compared with the control group, SH-SY5Y cells and HT22 cells treated with 4 and 8% sevoflurane for 12 h presented longer comet tails, in a concentration-dependent manner. However, the appearance of comet tails in SH-SY5Y cells and HT22 cells caused by sevoflurane were obviously prevented by pretreatment of NAC. (E,F) Statistical analysis of neutral comet assay demonstrated that sevoflurane induced a significant increase in the percentage of cells with comet tails and DNA content within the tails (p < 0.05, p < 0.01), which were markedly reversed by pretreatment of NAC (p < 0.05, p < 0.01). (G–L) Western blotting and quantitative analysis showed that 4 and 8% sevoflurane for 12 h concentration-dependently induced upregulation of DNA damage-related protein 8-OHdG, γH2AX, and p-ATM in SH-SY5Y cells, HT22 cells, and hippocampal neurons (p < 0.01). Compared with the control group, ^∗p < 0.05, ^∗∗p < 0.01. Data are represented as mean ± SD from five independent experiments.