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Applied and Environmental Microbiology, June 2007, p. 3958-3964, Vol. 73, No. 12
0099-2240/07/$08.00+0     doi:10.1128/AEM.02944-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Lactobacillus rhamnosus Strain GG Reduces Aflatoxin B1 Transport, Metabolism, and Toxicity in Caco-2 Cells{triangledown}

S. Gratz,1,2* Q. K. Wu,2 H. El-Nezami,2 R. O. Juvonen,3 H. Mykkänen,1 and P. C. Turner4

Department of Clinical Nutrition, University of Kuopio, P.O. Box 1627, 70211 Kuopio, Finland,1 Food and Health Research Centre, University of Kuopio, P.O. Box 1627, 70211 Kuopio, Finland,2 Department of Pharmacology and Toxicology, University of Kuopio, P.O. Box 1627, 70211 Kuopio, Finland,3 Molecular Epidemiology Unit, Centre for Epidemiology and Biostatistics, LIGHT Laboratories, University of Leeds, LS2 9JT Leeds, United Kingdom4

Received 19 December 2006/ Accepted 16 April 2007

The probiotic Lactobacillus rhamnosus GG is able to bind the potent hepatocarcinogen aflatoxin B1 (AFB1) and thus potentially restrict its rapid absorption from the intestine. In this study we investigated the potential of GG to reduce AFB1 availability in vitro in Caco-2 cells adapted to express cytochrome P-450 (CYP) 3A4, such that both transport and toxicity could be assessed. Caco-2 cells were grown as confluent monolayers on transmembrane filters for 21 days prior to all studies. AFB1 levels in culture medium were measured by high-performance liquid chromatography. In CYP 3A4-induced monolayers, AFB1 transport from the apical to the basolateral chamber was reduced from 11.1% ± 1.9% to 6.4% ± 2.5% (P = 0.019) and to 3.3% ± 1.8% (P = 0.002) within the first hour in monolayers coincubated with GG (1 x 1010 and 5 x 1010 CFU/ml, respectively). GG (1 x 1010 and 5 x 1010 CFU/ml) bound 40.1% ± 8.3% and 61.0% ± 6.0% of added AFB1 after 1 h, respectively. AFB1 caused significant reductions of 30.1% (P = 0.01), 49.4% (P = 0.004), and 64.4% (P < 0.001) in transepithelial resistance after 24, 48, and 72 h, respectively. Coincubation with 1 x 1010 CFU/ml GG after 24 h protected against AFB1-induced reductions in transepithelial resistance at both 24 h (P = 0.002) and 48 h (P = 0.04). DNA fragmentation was apparent in cells treated only with AFB1 cells but not in cells coincubated with either 1 x 1010 or 5 x 1010 CFU/ml GG. GG reduced AFB1 uptake and protected against both membrane and DNA damage in the Caco-2 model. These data are suggestive of a beneficial role of GG against dietary exposure to aflatoxin.

* Corresponding author. Mailing address: Department of Clinical Nutrition, University of Kuopio, P.O. Box 1627, 70211 Kuopio, Finland. Phone: 358-17 163615. Fax: 358-17 162792. E-mail: silvia.gratz{at}uku.fi

{triangledown} Published ahead of print on 20 April 2007.

Applied and Environmental Microbiology, June 2007, p. 3958-3964, Vol. 73, No. 12
0099-2240/07/$08.00+0     doi:10.1128/AEM.02944-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.





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