Midazolam impacts acetyl—And butyrylcholinesterase genes: An epigenetic explanation for postoperative delirium?
Fig 3
Methylation of BCHE in neuronal SH-SY5Y and U343 cells after midazolam incubation A) BCHE intron 2 methylation reduced after midazolam (50 μg/ml) exposure of SH-SY5Y cells (n = 3; p = 0.01) analyzed by methylation specific PCR. B) ELISA showed that histone H3 lysine 4 di-methylation (H3K4me2) decreased in U343 cells after incubation with 250 ng/ml midazolam (n = 3; p = 0.02) C) Chip-Assay confirmed binding of BCHE promoter region (90 bp) to H3K4me2; a 100 bp DNA Ladder was utilized; lanes 1, 7 show incubation with H3K27 antibody; lanes 2 and 6 show incubation with H3K4 antibody; lane 4 shows negative control without antibody and lane 5 shows positive control with RNA-polymerase II antibody (two experiments out of three are shown; n = 3). D) ACHE -571-/-670 promoter methylation was not affected by midazolam (50 μg/ml) exposure of SH-SY5Y cells (n = 3; p = n.s.) analyzed by methylation specific PCR. Data are presented as mean ± standard deviation.