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Cytotoxic, genotoxic, and oxidative stress-related effects of lysergic acid diethylamide (LSD) and phencyclidine (PCP) in the human neuroblastoma SH-SY5Y cell line

Andreja Jurič
Andreja Jurič
, 
Antonio Zandona
Antonio Zandona
, 
Blanka Tariba Lovaković
Blanka Tariba Lovaković
, 
Dubravka Rašić
Dubravka Rašić
, 
Alica Pizent
Alica Pizent
, 
Goran Kozina
Goran Kozina
, 
Maja Katalinić
Maja Katalinić
, 
Ana Lucić Vrdoljak
Ana Lucić Vrdoljak
 et 
Irena Brčić Karačonji
Irena Brčić Karačonji
   | 30 déc. 2021
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Article Category: Original article
Publié en ligne: 30 déc. 2021
Pages: 333 - 342
Reçu: 01 oct. 2021
Accepté: 01 nov. 2021
DOI: https://doi.org/10.2478/aiht-2021-72-3604
© 2021 Heinz-Elmar Tenorth, published by Sciendo
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

Figure 1

Frequency distribution of DNA damage in SH-SY5Y cells after treatment with LSD (6.25 μmol/L) and PCP (3.13 μmol/L) compared to control (untreated cells). *p<0.05 vs control (Pearson’s χ2 test)
Frequency distribution of DNA damage in SH-SY5Y cells after treatment with LSD (6.25 μmol/L) and PCP (3.13 μmol/L) compared to control (untreated cells). *p<0.05 vs control (Pearson’s χ2 test)

Figure 2

Concentration of malondialdehyde (MDA) measured in SH-SY5Y cells after treatment with LSD (6.25 μmol/L) and PCP (3.13 μmol/L) compared to control. *p<0.05 vs control (Mann-Whitney U test)
Concentration of malondialdehyde (MDA) measured in SH-SY5Y cells after treatment with LSD (6.25 μmol/L) and PCP (3.13 μmol/L) compared to control. *p<0.05 vs control (Mann-Whitney U test)

Figure 3

ROS production in SH-SY5Y cells measured with the DCFH-DA assay after treatment with LSD (6.25 μmol/L) and PCP (3.13 μmol/L) compared to control. *p<0.05 vs control (Mann-Whitney U test); DCFH-DA – 2’,7’-dichlorodihydrofluorescein diacetate
ROS production in SH-SY5Y cells measured with the DCFH-DA assay after treatment with LSD (6.25 μmol/L) and PCP (3.13 μmol/L) compared to control. *p<0.05 vs control (Mann-Whitney U test); DCFH-DA – 2’,7’-dichlorodihydrofluorescein diacetate

Figure 4

Glutathione (GSH) level measured in SH-SY5Y cells after treatment with LSD (6.25 μmol/L) and PCP (3.13 μmol/L) compared to control. *p<0.05 vs control (Mann-Whitney U test)
Glutathione (GSH) level measured in SH-SY5Y cells after treatment with LSD (6.25 μmol/L) and PCP (3.13 μmol/L) compared to control. *p<0.05 vs control (Mann-Whitney U test)

Figure 5

Glutathione peroxidase (GPx) activity measured in SH-SY5Y cells after treatment with LSD (6.25 μmol/L) and PCP (3.13 μmol/L) compared to control. *p<0.05 vs control (Mann-Whitney U test)
Glutathione peroxidase (GPx) activity measured in SH-SY5Y cells after treatment with LSD (6.25 μmol/L) and PCP (3.13 μmol/L) compared to control. *p<0.05 vs control (Mann-Whitney U test)

Figure 6

Superoxide dismutase (SOD) activity measured in SH-SY5Y cells after treatment with LSD (6.25 μmol/L) and PCP (3.13 μmol/L) compared to control. *p<0.05 vs control (Mann-Whitney U test)
Superoxide dismutase (SOD) activity measured in SH-SY5Y cells after treatment with LSD (6.25 μmol/L) and PCP (3.13 μmol/L) compared to control. *p<0.05 vs control (Mann-Whitney U test)

Figure 7

Catalase (CAT) activity measured in SH-SY5Y cells after treatment with LSD (6.25 μmol/L) and PCP (3.13 μmol/L) compared to control. *p<0.05 vs control (Mann-Whitney U test)
Catalase (CAT) activity measured in SH-SY5Y cells after treatment with LSD (6.25 μmol/L) and PCP (3.13 μmol/L) compared to control. *p<0.05 vs control (Mann-Whitney U test)

Primary DNA damage measured in SH-SY5Y cells using the alkaline comet assay after treatment with 6.25 μmol/L (2.02 mg/L) LSD, 3.13 μmol/L (0.76 mg/L) PCP, and in respective negative and positive controls

Sample Negative control (untreated cells) LSD PCP Positive control (H2O2)
Descriptor Tail DNA% Tail length (μm) Tail DNA% Tail length (μm) Tail DNA% Tail length (μm) Tail DNA% Tail length (μm)
Mean 1.57 13.55 3.14* 15.46* 2.30 15.65* 8.33* 15.84*
SE 0.14 0.26 0.33 0.32 0.25 0.27 0.66 0.39
SD 2.43 4.53 5.68 5.51 4.27 4.60 11.51 6.84
Median 0.22 12.50 0.62 13.75 0.21 14.58 2.87 13.75
Minimum 0 6.25 0 7.50 0 7.92 0 5.00
Maximum 11.97 28.33 45.77 54.17 29.28 37.08 63.81 45.83

Experimental schedule

Experimental group LSD (mg/L) PCP (mg/L)
0.39 μmol/L 0.13 0.09
0.78 μmol/L 0.25 0.19
1.56 μmol/L 0.50 0.38
3.13 μmol/L 1.01 0.76
6.25 μmol/L 2.02 1.52
12.50 μmol/L 4.04 3.04
25 μmol/L 8.09 6.08
50 μmol/L 16.17 12.17
100 μmol/L 32.34 24.34
Control untreated cells

Viability of SH-SY5Y cells exposed to LSD and PCP in concentrations of 0.39–100 μmol/L for 24 h determined with the MTS assay

Experimental group Viable cells (mean % ±SD)
LSD PCP
Control 99.67±3.74 99.20±4.42
0.39 μmol/L 97.77±7.94 92.65±6.25
0.78 μmol/L 93.68±8.35* 90.05±7.07
1.56 μmol/L 90.26±7.52* 86.54±11.08*
3.13 μmol/L 90.67±5.95* 84.17±9.03*
6.25 μmol/L 88.06±7.44* 78.10±5.49*
12.5 μmol/L 94.18±8.18 76.28±10.10*
25 μmol/L 100.17±7.29 75.57±9.15*
50 μmol/L 101.82±6.77 71.02±13.29*
100 μmol/L 98.09±7.63 72.80±8.85*
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