Effects of aflatoxin B1 on liver phase I and phase II enzymes induced in vitro on Sparus aurata hepatocytes primary culture

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Abstract

Among all know mycotoxins, aflatoxin B1 is one of the most studied for its hepatotoxic, carcinogenic, mutagenic, teratogenic, and immunosuppressant effects. However, metabolic and toxicological studies on aflatoxins in farmed Sparus aurata are limited and restricted to in vivo trials. This work aimed to study the effects of AFB1 acute and chronic exposure on CYP1A and GST enzymes induced in vitro on S. aurata hepatocytes by immunoblot analysis, thus relating the cytotoxic effects leading to cell death by apoptotic studies. Immunofluorescence analysis revealed that cell damage was not recoverable but permanent, as the cellular repair systems were unable to recover the induced toxic insult. Our results showed detection of several CYP1A bands, enlightening an indirect correlation between induction of CYP1A with dose and time of exposure. The decreased expression of CYP1A over prolonged exposure times, along with high toxic concentration, could be related with lethal damage observed on hepatocytes by contrast phase and immunofluorescence analysis. A particular pattern of expression was found for GST isoforms upon AFB1 exposure, identifying each isoform profile two different kind of toxic insult. The 65 KDa and the 49 KDa bands being suggestive for markers of acute and chronic response respectively. Interestingly, apoptosis induction, considered an early lesion to DNA, was found associated with the chronic damage along with the low toxic concentration. The new cell model from S. aurata has been proven to be a useful and valid tool to further investigate the modulated response of liver phase I and II enzymes to AFB1.

Autore Pugliese

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  • CASALINO E.;CENTODUCATI G.;SANTACROCE M.

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Anno di pubblicazione

2012

ISSN

1594-4077

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