Exposure of human neuroblastoma SH-SY5Y cells to amphetamine-type stimulants leads to oxidative-antioxidative imbalance associated with DNA damage and acetylcholine antagonism

Article

作者: Jurič, Andreja ; Zandona, Antonio ; Karačonji, Irena Brčić ; Canjuga, Irena ; Kozina, Goran ; Lovaković, Blanka Tariba ; Vrdoljak, Ana Lucić ; Kopjar, Nevenka ; Rešić, Arnes ; Katalinić, Maja ; Pizent, Alica ; Rašić, Dubravka ; Neuberg, Marijana

Amphetamine-type stimulants (ATSs) are widely abused substances that impair central and peripheral nervous system functions. The mechanisms of their toxicity on human neuronal cells have not been fully clarified yet but include effects on oxidative-antioxidative balance and interaction with synaptic enzymes/receptors. The aims of this study were to determine oxidant/antioxidant status, DNA integrity and the activity of neurotransmitter system components (monoamine oxidase A, MAO-A and nicotinic acetylcholine receptors, nAChR) in human neuroblastoma SH-SY5Y cells after 24-h exposure to different ATSs. In this matter, we first evaluated cell viability by MTS assay. After determination of the concentrations (near IC₂₅) suitable for conduction of the alkaline comet assay, these were further studied for the extent of DNA damage, along with the levels of malondialdehyde, reactive oxygen species (ROS), glutathione (GSH) and activities of antioxidative enzymes superoxide dismutase (SOD), glutathione peroxidase (GPx) and catalase (CAT). The activity of the MAO-A enzyme and nAChR activity were evaluated at concentration range 1-50 μM. Finally, the in silico pharmacokinetic parameters predictions of the tested ATSs were determined. When compared to untreated cells, the most notable result was obtained for amphetamine (AMP), where we observed a significant increase in ROS levels, SOD, GPx and CAT activity, and a decrease in GSH levels. Interestingly, all of the tested ATSs increased the activity of GPx, while the activities of the other enzymes were compound-dependent. The new psychoactive substance (NPS) mephedrone (4-MMC) had a similar effect as AMP, except that it did not affect CAT activity. At the tested concentrations, AMP, methamphetamine (METH) and 4-MMC also showed effects on DNA stability. None of the tested ATSs inhibited MAO-A in the tested concentration range, but AMP, METH, 3,4-methylenedioxymethamphetamine (MDMA) and 3,4-methylenedioxyamphetamine (MDA) inhibited acetylcholine activation of human nAChR. Taken together, significant induction of oxidative stress parameters, increased level of DNA damage detected and inhibition of nAChR activity indicate potential mechanisms of ATS substances action, and represent the direction for further studies to clarify the toxicological risks associated with ATS and/or NPS consumption.

2025-07-01·INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES

3D-printed Laponite/alginate hydrogel-based suppositories as innovative platforms for AT-MSC secretome delivery in IBD treatment

Article

作者: Igartua, Manoli ; Santos-Vizcaino, Edorta ; Hernandez, Rosa Maria ; Munoz-Perez, Elena

The secretome of Mesenchymal Stromal Cells (MSCs) has demonstrated effectiveness in the treatment of Inflammatory Bowel Disease (IBD), offering a safer and more predictable alternative to the direct administration of MSCs. While parenteral administration is common, rectal delivery of the secretome provides targeted treatment for localized conditions such as ulcerative colitis and proctitis. Alginate-Laponite 3D-printed suppositories (Alg-Lap 3DPS) have shown a remarkable versatility in their ability to encapsulate and deliver therapeutic agents during previous studies. These formulations are also adaptable in the release of molecules with different physicochemical properties, and a Bovine Serum Albumin (BSA) displacement strategy has been proposed in order to promote their disintegration and controlled release. As such, these 3DPS formulations represent highly versatile pharmaceutical forms, enabling the rectal delivery of diverse molecules with controlled release characteristics. In this study, we combined the therapeutic potential of Adipose tissue derived-MSCs-secretome (AT-S) with the versatility and efficacy of Alg-Lap 3DPS. Herein, we demonstrate that these formulations are well-suited for the encapsulation and release of AT-S, confirming their ability to release bioactive factors from AT-S such as Galectin-9 upon applying the displacement strategy. Moreover, we validate the bioactivity of the released factors in vitro by testing their effects on pro-inflammatory M1-polarized macrophages. Notably, this study not only confirms the anti-inflammatory capacity of the AT-S-loaded 3DPS in vitro but also aims to elucidate the underlying mechanisms driving these immunomodulatory effects, revealing the inherent immunomodulatory capacity of the Alg-Lap matrices themselves in M1 macrophages and establishing the AT-S-loaded 3DPS formulations as innovative therapeutic approaches for the treatment of IBD.

2025-06-01·Drug Testing and Analysis

Determination of amphetamines in hair samples using microextraction by packed sorbent and gas chromatography–mass spectrometry

Article

作者: Pires, Bruno ; Rosado, Tiago ; Barroso, Mário ; Gallardo, Eugenia ; Simão, Ana Y.

Abstract:

Several protocols for the analysis of amphetamine‐type stimulants (ATS) in hair have been developed over the years, with microextraction by packed sorbent (MEPS) being used for drugs like opiates, cocaine and ketamine. However, concerning ATS determination in hair samples, this approach has only been applied so far to amphetamine (AMP) and methamphetamine (MAMP). This study aimed at developing and validating a MEPS‐based procedure for the determination in hair of not only AMP and MAMP but also of 3,4‐methylenedioxyamphetamine (MDA), 3,4‐methylenedioxymethamphetamine (MDMA), 1‐(1,3‐benzodioxol‐5‐yl)propan‐2‐yl (ethyl)amine (MDE) and N‐methyl‐1‐(1,3‐benzodioxol‐5‐yl)‐2‐aminobutane (MBDB) as well.Hair, 50 mg, was incubated with 1 M sodium hydroxide (NaOH) at 45°C overnight, neutralization with 10 M hydrochloric acid (HCl) and centrifugation followed. The design of experiments approach was used for MEPS optimization, with the final optimized conditions including conditioning (250 μL methanol and deionized water), loading (18 × 100 μL) and elution (7 × 100 μL 2% NH4OH in acetonitrile). The eluted extract was evaporated to dryness and underwent microwave‐assisted derivatization with N‐methyl‐bis(trifluoroacetamide) (MBTFA), and it was afterwards injected onto the gas chromatography–mass spectrometer (GC–MS).The obtained recoveries ranged between 8% and 14% for AMP, 14% and 20% for MAMP, 10% and 15% for MDA, 18% and 28% for MDMA, 25% and 43% for MDE and 34% and 52% for MBDB, and the method was linear from 0.2 to 5.0 ng/mg. Precision and accuracy were in accordance with international method validation guidelines. This novel method involving MEPS coupled to GC–MS offers a swift, eco‐friendly and cost‐effective alternative to traditional procedures for detecting these AMPs in hair samples.

100 项与 Antistasin(Merck Sharp & Dohme Corp.) 相关的药物交易

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