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​The effect of T-2 toxin on cell metabolism, oxidative stress and ER stress in 3D hepatic cell cultures of chicken origin
van Eijk Victoria Elizabeth Apollonia - year 5
University of Veterinary Medicine Budapest, Department of Physiology and Biochemistry
Supervisors: Dr. Zsuzsanna Neogrády, Júlia Vörösházi

Abstract:

T-2 toxin is a trichothecene mycotoxin produced by different Fusarium species that often contaminate cereal grains and their processed food products. Since poultry species consume mainly cereals, they are at a greater risk of T-2 toxin exposure. The toxin has genotoxic, cytotoxic, and immunomodulatory effects in poultry, leading to reduced productivity. The main cellular effect of the toxin is the inhibition of protein synthesis. It also induces oxidative stress that could lead to endoplasmic reticulum (ER) stress. Therefore, the investigation of the cellular effects of T-2 toxin is highly important to agriculture and veterinary medicine.

In this study we examined the cellular effects of T-2 toxin in three-dimensional (3D) hepatocyte – non-parenchymal (NP) cell co-cultures of chicken origin. The co-cultures were exposed to three concentrations (100, 500, and 1000 nM) of T-2 toxin for different incubation times (24 and 48 h). The metabolic activity of the cells was determined by CCK-8 (Cell Counting Kit-8) test. To assess the oxidative stress and ER stress, the amount of protein carbonyl (PC) was measured from cell lysates, while malondialdehyde (MDA) as well as glucose regulated protein 78 (GRP78) and heat shock protein 27 (HSP27) were measured from medium using chicken-specific ELISA tests.

Our results showed that T-2 toxin significantly decreased the metabolic activity in every treatment group after both incubations. Metabolic adaptation was indicated by the liver cells as metabolic depression was alleviated after the longer incubation period. The MDA release of the cells was significantly decreased only in the 1000 nM treatment group after 24 h, while after 48 h all three concentrations of T-2 toxin significantly decreased the MDA production of the cell cultures. The higher levels of T-2 toxin significantly decreased the PC concentration of the cultured cells after 48 h. No significant changes were observed in GRP78 concentration of the cell cultures. In the case of HSP27, all T-2 toxin treatments significantly decreased concentration after 48 h.

These findings suggest that T-2 toxin triggered a mild metabolic depression of the hepatic cells, but in our 3D co-culture model that mimics better the in vivo conditions, the toxin could also induce protective mechanisms within the cells resulting in the reduction of oxidative and by chance ER stress when applied in higher concentrations for longer periods of time.



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