Abstract
Many pesticides are used increasingly in combinations during crop protection and their stability ensures the presence of such combinations in foodstuffs. The effects of three fungicides, pyrimethanil, cyprodinil and fludioxonil, were investigated together and separately on U251 and SH-SY5Y cells, which can be representative of human CNS glial and neuronal cells respectively. Over 48h, all three agents showed significant reductions in cellular ATP, at concentrations that were more than tenfold lower than those which significantly impaired cellular viability. The effects on energy metabolism were reflected in their marked toxic effects on mitochondrial membrane potential. In addition, evidence of oxidative stress was seen in terms of a fall in cellular thiols coupled with increases in the expression of enzymes associated with reactive species formation, such as GSH peroxidase and superoxide dismutase. The glial cell line showed significant responsiveness to the toxin challenge in terms of changes in antioxidant gene expression, although the neuronal SH-SY5Y line exhibited greater vulnerability to toxicity, which was reflected in significant increases in caspase-3 expression, which is indicative of the initiation of apoptosis. Cyprodinil was the most toxic agent individually, although oxidative stress-related enzyme gene expression increases appeared to demonstrate some degree of synergy in the presence of the combination of agents. This report suggests that the impact of some pesticides, both individually and in combinations, merits further study in terms of their impact on human cellular health.
Highlights
Pesticide use is integral in world agriculture
The process of wine production generally reduces pesticide contamination in comparison with that of grapes, this depends on the physiochemical properties of the agents used [9] and it is apparent that wines sourced from Europe contain multiple residues of many different pesticide agents in measurable quantities [12,13]
We have explored the effects of a combination of three commonly used biocides on cell viability, mitochondrial health and generation of oxidative stress, alone and in combination
Summary
Pesticide use is integral in world agriculture. In the Californian wine industry, over twenty thousand tonnes of pesticide are used per annum, including more than 150 different agents [1]. Considerable research has been carried out on the impact of pesticides on various non-target flora and fauna [17,18] and there is evidence that many biocidal agents lack specificity They can exert significant mammalian toxicity that may be synergistic where combinations of different agents are deployed [19,20,21]. We have utilized two cell lines, gliomal U251 and neuroblastomal SH-SY5Y cells, which model the basic cell types of the human central nervous system, the astrocytes and neurones In this preliminary study, we have explored the effects of a combination of three commonly used biocides (pyrimethanil, ciprodinil and fludioxonil) on cell viability, mitochondrial health and generation of oxidative stress, alone and in combination
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