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. 2019 May 3;43:473–486. doi: 10.1016/j.ebiom.2019.04.048

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© 2019 The Authors

This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

PMC Copyright notice

Fig. 1 — Sevoflurane reduced the expression of TYMS and ERMN in both macaque and mice prefrontal cortex and was associated with the reduction of folate levels in blood in children.

a. RNA sequencing assay indicated that the sevoflurane anesthesia induced the downregulation of TYMS and ERMN in the cortex of rhesus macaque (N =2).

b. The sevoflurane anesthesia decreased the mRNA level of TYMS detected by quantitative RT-PCR (N = 3). The data shown are the mean ± SD, p= 0.013, Student's t-test.

c. The whole genome methylation analysis showed that there was higher ERMN promoter methylation in the cortex of the rhesus macaque after the sevoflurane anesthesia as compared to that in the control condition (N = 2).

d. qRT-PCR assay confirmed the downregulation of ERMN mRNA levels in cortex tissues of rhesus macaque following the sevoflurane anesthesia as compared to those in the cortex tissues of rhesus macaque following the control condition (N = 3). The data shown are the mean ± SD, p = 0.0013, Student's t-test.

The down-regulation of ERMN (e) and TYMS (f) was detected by qPCR in the prefrontal cortex of P8 mice following the sevoflurane anesthesia as compared to the control condition. The data shown are the mean ± SD, p = 0.007 and p = 0.014, Student's t-test. Representative pictures showed that the sevoflurane anesthesia reduced the protein levels of TYMS (g) and ERMN (h), detected by Western blot analysis in the prefrontal cortex of P8 mice (N = 3).

The quantification of the Western blot in g and h showed that the sevoflurane anesthesia decreased the protein levels of TYMS (i, p = 0.022, N = 3, Student's t-test) and ERMN (j, p = 0.035, N = 3, Student's t-test).

k. There were postoperative reductions in the blood levels of folate in the participants who had surgery under sevoflurane anesthesia: (median: 14.50; interquartile range: 8.65, 17.33) versus (median: 16.40; interquartile range: 9.32, 17.75) (p < 0.0001, N = 20, Mann–Whitney U test).

The data shown are the mean ± SD, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. The Wilcoxon-Mann-Whitney test used for the analysis of the data that were not normally distributed for the comparison of two samples. A two-sided Student's t-test was used for the analysis of the data that were normally distributed for the comparison of two samples.

Control: ctrl.; sevoflurane: sevo.; ERM-like protein: ERMN; thymidylate synthase: TYMS.