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Published on: December 11, 2020
Oxidative stress and gene expression profiling of cell death pathways in alpha-cypermethrin-treated SH-SY5Y cells
Alejandro Romero1, Eva Ramos1, Irma Ares1
1Department of Toxicology and Pharmacology, Faculty of Veterinary Medicine, Universidad Complutense de Madrid, 28040, Madrid, Spain.
Abstract:
In this study, we investigated the induction of oxidative stress and apoptosis in human neuroblastoma cell line SH-SY5Y in response to alpha-cypermethrin (α-CYPER) exposure. MTT and LDH assays were carried out to assess the α-CYPER cytotoxicity. The IC50 value for α-CYPER was calculated to be 78.3 ± 2.98 µM for the MTT assay and 71.5 ± 3.94 µM for LDH assay. The pyrethroid α-CYPER (1-100 µM), in a dose-dependent manner, induced a significant increase in lipid peroxides measured as malondialdehyde (MDA) and in the levels of nitric oxide (NO). The neuroprotective role of three antioxidants, melatonin (MEL), Trolox and N-acetylcysteine (NAC) against α-CYPER-induced oxidative stress was examined. Compared to other antioxidants, MEL (1 µM) treatment showed the most effective protection against α-CYPER-induced lipid peroxidation and NO production. The effects of α-CYPER on gene expression profiling of cell death pathway in human neuroblastoma SH-SY5Y cells were also investigated. Of the 84 genes examined (P < 0.001; fold change >1.5), changes in mRNA levels were detected in 39 genes: 36 were up-regulated and 3 were down-regulated. A greater fold change reversion than 3.5-fold was observed on the up-regulated ATP6V1G2, BCL2, CASP9, FAS, GADD45A, SPATA2, SYCP2, ATG7, NFKB1, SNCA, ULK1 and JPH3 genes. The results demonstrated that α-CYPER alters the expression of apoptosis-, autophagy- and necrosis genes as well as induces oxidative stress which may lead to DNA damage. The detailed knowledge of the changes in gene expression obtained will provide a basis for further elucidating the molecular mechanisms of the α-CYPER-induced toxicity.
Insights
Alpha-cypermethrin (α-CYPER) induces oxidative stress and apoptosis in human neuroblastoma cells. Melatonin effectively protected against α-CYPER toxicity by reducing lipid peroxidation and nitric oxide levels.
Area of Science:
- Toxicology
- Neuroscience
- Molecular Biology
Background:
- Alpha-cypermethrin (α-CYPER) is a widely used pyrethroid insecticide.
- Neuroblastoma cells are a relevant model for studying neurotoxic effects.
- Understanding the molecular mechanisms of α-CYPER toxicity is crucial for risk assessment.
Purpose of the Study:
- To investigate the induction of oxidative stress and apoptosis in SH-SY5Y cells by α-CYPER.
- To evaluate the neuroprotective effects of antioxidants against α-CYPER-induced toxicity.
- To analyze the impact of α-CYPER on the gene expression profile of cell death pathways.
Main Methods:
- Cytotoxicity was assessed using MTT and LDH assays to determine IC50 values.
- Lipid peroxidation (MDA) and nitric oxide (NO) levels were measured to quantify oxidative stress.
- Gene expression profiling was performed to identify changes in cell death pathway-related genes.
Main Results:
- α-CYPER exposure led to a dose-dependent increase in oxidative stress markers (MDA and NO) and induced apoptosis.
- Melatonin (MEL) demonstrated significant neuroprotective effects, more potent than Trolox and N-acetylcysteine (NAC).
- α-CYPER altered the expression of 39 genes involved in apoptosis, autophagy, and necrosis, with notable changes in genes like BCL2, CASP9, and ATG7.
Conclusions:
- α-CYPER induces oxidative stress and apoptosis in human neuroblastoma cells.
- Melatonin shows promising neuroprotective potential against α-CYPER toxicity.
- α-CYPER exposure significantly impacts the expression of genes related to programmed cell death and cellular stress, potentially leading to DNA damage.
