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Cyanobacterial lipopeptides puwainaphycins and minutissamides induce disruptive and pro-inflammatory processes in Caco-2 human intestinal barrier model

O. Vašíček, J. Hájek, L. Bláhová, P. Hrouzek, P. Babica, L. Kubala, L. Šindlerová

. 2020 ; 96 (-) : 101849. [pub] 20200612

Language English Country Netherlands

Document type Journal Article, Research Support, Non-U.S. Gov't

Persistent link   https://www.medvik.cz/link/bmc21012524

Puwainaphycins (PUW) and minutissamides (MIN) are cyanobacterial lipopeptides found in various cyanobacterial species. The first possible target of human exposure to them is intestinal epithelium but effect of PUW/MIN on enterocytes is not known at all. Using differentiated Caco-2 cells, PUW F was found to be cytotoxic from 5 µM concentration based on lactate dehydrogenase release assay and total protein concentration. However, it is also able to induce production of interleukin 8 in non-cytotoxic concentrations 1 and 2.5 µM detected by ELISA. Effects of MIN A and C were similar but less pronounced compared to PUW F. On the other hand, MIN D was the least toxic compound with no significant pro-inflammatory effects. Surprisingly, pro-inflammatory activation of the cells by PUW F and MIN C resulted in an increase in tight junction (TJ) protein claudin 4 expression determined by western blot analysis and confirmed by confocal microscopy. Furthermore, decrease in expression of zonula occludens 3, another TJ protein, was observed after the exposure to PUW F. Taken together, these cytotoxic lipopeptides, especially PUW F, are to be studied more deeply due to their capability to activate and/or deregulate human enterocytes in low concentrations.

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$a Puwainaphycins (PUW) and minutissamides (MIN) are cyanobacterial lipopeptides found in various cyanobacterial species. The first possible target of human exposure to them is intestinal epithelium but effect of PUW/MIN on enterocytes is not known at all. Using differentiated Caco-2 cells, PUW F was found to be cytotoxic from 5 µM concentration based on lactate dehydrogenase release assay and total protein concentration. However, it is also able to induce production of interleukin 8 in non-cytotoxic concentrations 1 and 2.5 µM detected by ELISA. Effects of MIN A and C were similar but less pronounced compared to PUW F. On the other hand, MIN D was the least toxic compound with no significant pro-inflammatory effects. Surprisingly, pro-inflammatory activation of the cells by PUW F and MIN C resulted in an increase in tight junction (TJ) protein claudin 4 expression determined by western blot analysis and confirmed by confocal microscopy. Furthermore, decrease in expression of zonula occludens 3, another TJ protein, was observed after the exposure to PUW F. Taken together, these cytotoxic lipopeptides, especially PUW F, are to be studied more deeply due to their capability to activate and/or deregulate human enterocytes in low concentrations.
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$a Hájek, Jan $u Institute of Microbiology, Centre Algatech, The Czech Academy of Sciences, Novohradska 237, Trebon 379 80, Czech Republic. Electronic address: hajek@alga.cz
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$a Bláhová, Lucie $u RECETOX, Faculty of Science, Masaryk University, Kamenice 753/5, Brno 625 00, Czech Republic. Electronic address: lucie.blahova@recetox.muni.cz
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$a Hrouzek, Pavel $u Institute of Microbiology, Centre Algatech, The Czech Academy of Sciences, Novohradska 237, Trebon 379 80, Czech Republic. Electronic address: hrouzek@alga.cz
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$a Babica, Pavel $u RECETOX, Faculty of Science, Masaryk University, Kamenice 753/5, Brno 625 00, Czech Republic; Institute of Botany, The Czech Academy of Sciences, Lidická 25/27, Brno 602 00, Czech Republic. Electronic address: pavel.babica@recetox.muni.cz
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$a Kubala, Lukáš $u Institute of Biophysics of the Czech Academy of Sciences, Kralovopolska 135, Brno 612 65, Czech Republic. Electronic address: kubalal@ibp.cz
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$a Šindlerová, Lenka $u Institute of Biophysics of the Czech Academy of Sciences, Kralovopolska 135, Brno 612 65, Czech Republic. Electronic address: sindler@ibp.cz
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