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Cytotoxicity-related effects of imidazolium and chlorinated bispyridinium oximes in SH-SY5Y cells

Antonio Zandona

Antonio Zandona

Institute for Medical Research and Occupational HealthZagreb, Croatia
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Zandona, Antonio
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Tamara Zorbaz

Tamara Zorbaz

Institute for Medical Research and Occupational HealthZagreb, Croatia
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Zorbaz, Tamara
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Katarina Miš

Katarina Miš

University of Ljubljana Faculty of Medicine, Institute of PathophysiologyLjubljana, Slovenia
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Miš, Katarina
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Sergej Pirkmajer

Sergej Pirkmajer

University of Ljubljana Faculty of Medicine, Institute of PathophysiologyLjubljana, Slovenia
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Pirkmajer, Sergej
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Maja Katalinić

Maja Katalinić

Institute for Medical Research and Occupational HealthZagreb, Croatia
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Katalinić, Maja
  
30 dic 2022
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Categoria dell'articolo: Original article
Pubblicato online: 30 dic 2022
Pagine: 277 - 284
Ricevuto: 01 ott 2022
Accettato: 01 dic 2022
DOI: https://doi.org/10.2478/aiht-2022-73-3688
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© 2022 Antonio Zandona, Tamara Zorbaz, Katarina Miš, Sergej Pirkmajer, and Maja Katalinić, published by Sciendo
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.

Figure 1

Structures of oximes tested in this study
Structures of oximes tested in this study

Figure 2

ATP in SH-SY5Y cells after 4 h exposure to selected oximes expressed as the percentage of control (untreated cells). Oxime concentrations: K867 (400 μM), K870 (15 μM), VII (250 μM) and X (200 μM). #p≤0.01; *p≤0.0001
ATP in SH-SY5Y cells after 4 h exposure to selected oximes expressed as the percentage of control (untreated cells). Oxime concentrations: K867 (400 μM), K870 (15 μM), VII (250 μM) and X (200 μM). #p≤0.01; *p≤0.0001

Figure 3

Time-dependent effects of the selected oximes K867 (400 μmol/L), K870 (15 μmol/L), VII (250 μmol/L), and X (200 μmol/L) on intracellular signalling in serum-starved SH-SY5Y cells over 5–60 min. Position towards bends and molecular weight markers in kDa are indicated on the right side of the blots and Ponceau S. Results are presented as means ± SD (n=9) &p≤0.05; #p≤0.01; $p≤0.001; *p≤0.0001. B– baseline; pERK1/2 – phospho-ERK1/2 (Thr202/Tyr204); pNFkB – phospho-NFκbeta p65 (Ser536); p-p38MAPK – phospho-p38MAPK (Thr180/Tyr182); p-STAT3 – phospho-STAT3 (Tyr705); p-ACC – phospho-ACC (Ser79); p-AMPKα – phospho-AMPKalpha (Thr172); t– time
Time-dependent effects of the selected oximes K867 (400 μmol/L), K870 (15 μmol/L), VII (250 μmol/L), and X (200 μmol/L) on intracellular signalling in serum-starved SH-SY5Y cells over 5–60 min. Position towards bends and molecular weight markers in kDa are indicated on the right side of the blots and Ponceau S. Results are presented as means ± SD (n=9) &p≤0.05; #p≤0.01; $p≤0.001; *p≤0.0001. B– baseline; pERK1/2 – phospho-ERK1/2 (Thr202/Tyr204); pNFkB – phospho-NFκbeta p65 (Ser536); p-p38MAPK – phospho-p38MAPK (Thr180/Tyr182); p-STAT3 – phospho-STAT3 (Tyr705); p-ACC – phospho-ACC (Ser79); p-AMPKα – phospho-AMPKalpha (Thr172); t– time

Figure 4

Target class frequencies of oximes K867, K870, VII, and X predicted by the SwissTargetPrediction engine. The dashed frame highlights the family of G-protein-coupled receptors as the most probable target of interaction
Target class frequencies of oximes K867, K870, VII, and X predicted by the SwissTargetPrediction engine. The dashed frame highlights the family of G-protein-coupled receptors as the most probable target of interaction

Figure 5

iCa2+ levels in SH-SY5Y cells before and after the addition of ACh (5 μmol/L) in the presence of oximes K867 (400 μmol/L), K870 (25 μmol/L), VII (250 μmol/L), and X (200 μmol/L) compared to control (not treated with the oximes)
iCa2+ levels in SH-SY5Y cells before and after the addition of ACh (5 μmol/L) in the presence of oximes K867 (400 μmol/L), K870 (25 μmol/L), VII (250 μmol/L), and X (200 μmol/L) compared to control (not treated with the oximes)

Primary antibodies (Cell Signaling Technology Inc, USA) used for Western blotting

Antibody Cat. No. Host Type Dilution
Anti p-ERK (Thr202/Tyr204) #4370 Rabbit Monoclonal 1:2000
Anti pNFκB p65 (Ser536) #3033 Rabbit Monoclonal 1:2000
Anti p-p38 MAPK (Thr180/Tyr182) #4511 Rabbit Monoclonal 1:1000
Anti p-STAT3 (Tyr705) #9145 Rabbit Monoclonal 1:1000
Anti p-ACC (Ser79) #3661 Rabbit Polyclonal 1:1000
Anti p-AMPKα (Thr172) #2535 Rabbit Monoclonal 1:1000
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