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Early Activation of MAPK p44/42 Is Partially Involved in DON-Induced Disruption of the Intestinal Barrier Function and Tight Junction Network

View Article: PubMed Central - PubMed

ABSTRACT

Deoxynivalenol (DON), produced by the plant pathogens Fusariumgraminearum and Fusarium culmorum, is one of the most common mycotoxins, contaminating cereal and cereal-derived products. Although worldwide contamination of food and feed poses health threats to humans and animals, pigs are particularly susceptible to this mycotoxin. DON derivatives, such as deepoxy-deoxynivalenol (DOM-1), are produced by bacterial transformation of certain intestinal bacteria, which are naturally occurring or applied as feed additives. Intestinal epithelial cells are the initial barrier against these food- and feed-borne toxins. The present study confirms DON-induced activation of MAPK p44/42 and inhibition of p44/42 by MAPK-inhibitor U0126 monoethanolate. Influence of DON and DOM-1 on transepithelial electrical resistance (TEER), viability and expression of seven tight junction proteins (TJ), as well as the potential of U0126 to counteract DON-induced effects, was assessed. While DOM-1 showed no effect, DON significantly reduced TEER of differentiated IPEC-J2 and decreased expression of claudin-1 and -3, while leaving claudin-4; ZO-1, -2, and -3 and occludin unaffected. Inhibition of p44/42 counteracted DON-induced TEER decrease and restored claudin-3, but not claudin-1 expression. Therefore, effects of DON on TEER and claudin-3 are at least partially p44/42 mediated, while effects on viability and claudin-1 are likely mediated via alternative pathways.

No MeSH data available.


Related in: MedlinePlus

Progression of TEER (transepithelial electrical resistance) of intestinal porcine epithelial cells (IPEC-J2) seeded in 1.12 cm2 Transwell® polyester membrane inserts, measured at indicated time points over a period of 30 days. Cells reached a steady-state TEER phase between Days 7 and 11. Data represent mean ± standard deviation (SD), n = 4.
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toxins-08-00264-f001: Progression of TEER (transepithelial electrical resistance) of intestinal porcine epithelial cells (IPEC-J2) seeded in 1.12 cm2 Transwell® polyester membrane inserts, measured at indicated time points over a period of 30 days. Cells reached a steady-state TEER phase between Days 7 and 11. Data represent mean ± standard deviation (SD), n = 4.

Mentions: To monitor monolayer formation, the progression of TEER of IPEC-J2, cultured in 1.12 cm2 Transwell® polyester membrane inserts, was recorded over a period of 30 days. IPEC-J2 reached a resistance maximum after seven days of cultivation. At this time point, a well-established IPEC-J2 monolayer was observed microscopically. Between Day 7 (9.52 ± 0.15 kOhm × cm2) and Day 11 (8.88 ± 0.18 kOhm × cm2), TEER reached a steady-state phase (Figure 1), indicating a state of differentiation, suitable for toxin treatment.


Early Activation of MAPK p44/42 Is Partially Involved in DON-Induced Disruption of the Intestinal Barrier Function and Tight Junction Network
Progression of TEER (transepithelial electrical resistance) of intestinal porcine epithelial cells (IPEC-J2) seeded in 1.12 cm2 Transwell® polyester membrane inserts, measured at indicated time points over a period of 30 days. Cells reached a steady-state TEER phase between Days 7 and 11. Data represent mean ± standard deviation (SD), n = 4.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC5037490&req=5

toxins-08-00264-f001: Progression of TEER (transepithelial electrical resistance) of intestinal porcine epithelial cells (IPEC-J2) seeded in 1.12 cm2 Transwell® polyester membrane inserts, measured at indicated time points over a period of 30 days. Cells reached a steady-state TEER phase between Days 7 and 11. Data represent mean ± standard deviation (SD), n = 4.
Mentions: To monitor monolayer formation, the progression of TEER of IPEC-J2, cultured in 1.12 cm2 Transwell® polyester membrane inserts, was recorded over a period of 30 days. IPEC-J2 reached a resistance maximum after seven days of cultivation. At this time point, a well-established IPEC-J2 monolayer was observed microscopically. Between Day 7 (9.52 ± 0.15 kOhm × cm2) and Day 11 (8.88 ± 0.18 kOhm × cm2), TEER reached a steady-state phase (Figure 1), indicating a state of differentiation, suitable for toxin treatment.

View Article: PubMed Central - PubMed

ABSTRACT

Deoxynivalenol (DON), produced by the plant pathogens Fusariumgraminearum and Fusarium culmorum, is one of the most common mycotoxins, contaminating cereal and cereal-derived products. Although worldwide contamination of food and feed poses health threats to humans and animals, pigs are particularly susceptible to this mycotoxin. DON derivatives, such as deepoxy-deoxynivalenol (DOM-1), are produced by bacterial transformation of certain intestinal bacteria, which are naturally occurring or applied as feed additives. Intestinal epithelial cells are the initial barrier against these food- and feed-borne toxins. The present study confirms DON-induced activation of MAPK p44/42 and inhibition of p44/42 by MAPK-inhibitor U0126 monoethanolate. Influence of DON and DOM-1 on transepithelial electrical resistance (TEER), viability and expression of seven tight junction proteins (TJ), as well as the potential of U0126 to counteract DON-induced effects, was assessed. While DOM-1 showed no effect, DON significantly reduced TEER of differentiated IPEC-J2 and decreased expression of claudin-1 and -3, while leaving claudin-4; ZO-1, -2, and -3 and occludin unaffected. Inhibition of p44/42 counteracted DON-induced TEER decrease and restored claudin-3, but not claudin-1 expression. Therefore, effects of DON on TEER and claudin-3 are at least partially p44/42 mediated, while effects on viability and claudin-1 are likely mediated via alternative pathways.

No MeSH data available.


Related in: MedlinePlus