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Regulation of epithelial cell tight junctions by protease-activated receptor 2.

Enjoji S, Ohama T, Sato K - J. Vet. Med. Sci. (2014)

Bottom Line: Although the application of a PAR2-activating peptide, PAR2-AP, from the apical side of MDCK cells failed to modify the transepithelial resistance (TER), its application from the basal side markedly suppressed the TER.A p38 MAP kinase inhibitor, SB202190, inhibited PAR2-AP-induced changes in TER.Our results suggest that the activation of PAR2 leads to the disruption of tight junctions and increases the barrier permeability through the activation of p38 MAPK, which may cause the initiation and exacerbation of inflammation.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Veterinary Pharmacology, Joint Faculty of Veterinary Medicine, Yamaguchi University, 1677-1 Yoshida, Yamaguchi 753-8515, Japan.

ABSTRACT
A layer of epithelial cells prevents the invasion of bacteria and the entry of foreign substances into the underlying tissue. The disruption of epithelial tight junctions initiates and exacerbates inflammation. However, the precise mechanism underlying the disruption of the epithelial tight junction remains unclear. The activation of protease-activated receptor 2 (PAR2) by serine proteases produced by some bacteria and mast cells contributes to inflammation in many tissues. In the present study, we tested the hypothesis that PAR2 activation affects the structure and function of tight junctions in Madin-Darby canine kidney (MDCK) cells. Although the application of a PAR2-activating peptide, PAR2-AP, from the apical side of MDCK cells failed to modify the transepithelial resistance (TER), its application from the basal side markedly suppressed the TER. In 3-dimensional cultures of MDCK cells expressing the mCherry-tagged PAR2, a lateral localization of PAR2 was observed. The application of PAR2-AP from the basal side changed the localization of the tight junctional protein, zonula occludin-1. Furthermore, PAR2-AP induced the phosphorylation of p38 MAP kinase. A p38 MAP kinase inhibitor, SB202190, inhibited PAR2-AP-induced changes in TER. Our results suggest that the activation of PAR2 leads to the disruption of tight junctions and increases the barrier permeability through the activation of p38 MAPK, which may cause the initiation and exacerbation of inflammation.

Show MeSH
Application of PAR2-activating peptide (PAR2-AP) from the basal side of Madin-Darbycanine kidney (MDCK) cells decreases transepithelial electrical resistance (TER). TERacross MDCK cells was measured following stimulation without (Control) or with PAR2-AP(100 µM) from the apical side (Apical) or the basal side (Basal) forindicated periods. TER of MDCK cells at 0 hr was considered 100%.*P<0.05 vs. Control. N=3.
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fig_001: Application of PAR2-activating peptide (PAR2-AP) from the basal side of Madin-Darbycanine kidney (MDCK) cells decreases transepithelial electrical resistance (TER). TERacross MDCK cells was measured following stimulation without (Control) or with PAR2-AP(100 µM) from the apical side (Apical) or the basal side (Basal) forindicated periods. TER of MDCK cells at 0 hr was considered 100%.*P<0.05 vs. Control. N=3.

Mentions: We examined the effect of PAR2 stimulation on TER, a major marker of tight junctionformation, in MDCK cells. A PAR2-activating peptide (PAR2-AP, 100 µM) wasapplied onto the cell culture insert (apical side of MDCK) or into the well (basal side ofMDCK). Although the stimulation with PAR2 from the apical side did not alter the TER, thestimulation from the basal side significantly suppressed the TER after 30 min (Fig. 1Fig. 1.


Regulation of epithelial cell tight junctions by protease-activated receptor 2.

Enjoji S, Ohama T, Sato K - J. Vet. Med. Sci. (2014)

Application of PAR2-activating peptide (PAR2-AP) from the basal side of Madin-Darbycanine kidney (MDCK) cells decreases transepithelial electrical resistance (TER). TERacross MDCK cells was measured following stimulation without (Control) or with PAR2-AP(100 µM) from the apical side (Apical) or the basal side (Basal) forindicated periods. TER of MDCK cells at 0 hr was considered 100%.*P<0.05 vs. Control. N=3.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4197149&req=5

fig_001: Application of PAR2-activating peptide (PAR2-AP) from the basal side of Madin-Darbycanine kidney (MDCK) cells decreases transepithelial electrical resistance (TER). TERacross MDCK cells was measured following stimulation without (Control) or with PAR2-AP(100 µM) from the apical side (Apical) or the basal side (Basal) forindicated periods. TER of MDCK cells at 0 hr was considered 100%.*P<0.05 vs. Control. N=3.
Mentions: We examined the effect of PAR2 stimulation on TER, a major marker of tight junctionformation, in MDCK cells. A PAR2-activating peptide (PAR2-AP, 100 µM) wasapplied onto the cell culture insert (apical side of MDCK) or into the well (basal side ofMDCK). Although the stimulation with PAR2 from the apical side did not alter the TER, thestimulation from the basal side significantly suppressed the TER after 30 min (Fig. 1Fig. 1.

Bottom Line: Although the application of a PAR2-activating peptide, PAR2-AP, from the apical side of MDCK cells failed to modify the transepithelial resistance (TER), its application from the basal side markedly suppressed the TER.A p38 MAP kinase inhibitor, SB202190, inhibited PAR2-AP-induced changes in TER.Our results suggest that the activation of PAR2 leads to the disruption of tight junctions and increases the barrier permeability through the activation of p38 MAPK, which may cause the initiation and exacerbation of inflammation.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Veterinary Pharmacology, Joint Faculty of Veterinary Medicine, Yamaguchi University, 1677-1 Yoshida, Yamaguchi 753-8515, Japan.

ABSTRACT
A layer of epithelial cells prevents the invasion of bacteria and the entry of foreign substances into the underlying tissue. The disruption of epithelial tight junctions initiates and exacerbates inflammation. However, the precise mechanism underlying the disruption of the epithelial tight junction remains unclear. The activation of protease-activated receptor 2 (PAR2) by serine proteases produced by some bacteria and mast cells contributes to inflammation in many tissues. In the present study, we tested the hypothesis that PAR2 activation affects the structure and function of tight junctions in Madin-Darby canine kidney (MDCK) cells. Although the application of a PAR2-activating peptide, PAR2-AP, from the apical side of MDCK cells failed to modify the transepithelial resistance (TER), its application from the basal side markedly suppressed the TER. In 3-dimensional cultures of MDCK cells expressing the mCherry-tagged PAR2, a lateral localization of PAR2 was observed. The application of PAR2-AP from the basal side changed the localization of the tight junctional protein, zonula occludin-1. Furthermore, PAR2-AP induced the phosphorylation of p38 MAP kinase. A p38 MAP kinase inhibitor, SB202190, inhibited PAR2-AP-induced changes in TER. Our results suggest that the activation of PAR2 leads to the disruption of tight junctions and increases the barrier permeability through the activation of p38 MAPK, which may cause the initiation and exacerbation of inflammation.

Show MeSH