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Effect of factor XIII on endothelial barrier function.

Noll T, Wozniak G, McCarson K, Hajimohammad A, Metzner HJ, Inserte J, Kummer W, Hehrlein FW, Piper HM - J. Exp. Med. (1999)

Bottom Line: The thrombin-activated plasma factor XIII (1 U/ml) reduced albumin permeability of endothelial monolayers within 20 min by 30 +/- 7% (basal value of 5.9 +/- 0.4 x 10(-6) cm/s), whereas the nonactivated plasma factor XIII had no effect.This study shows that activated factor XIII reduces endothelial permeability.Its effect seems related to a modification of the paracellular passageways in endothelial monolayers.

View Article: PubMed Central - PubMed

Affiliation: Physiologisches Institut, Justus-Liebig-Universität, D-35392 Giessen, Germany. thomas.noll@physiologie.med.uni-giessen.de

ABSTRACT
The effect of factor XIII on endothelial barrier function was studied in a model of cultured monolayers of porcine aortic endothelial cells and saline-perfused rat hearts. The thrombin-activated plasma factor XIII (1 U/ml) reduced albumin permeability of endothelial monolayers within 20 min by 30 +/- 7% (basal value of 5.9 +/- 0.4 x 10(-6) cm/s), whereas the nonactivated plasma factor XIII had no effect. Reduction of permeability to the same extent, i.e., by 34 +/- 9% could be obtained with the thrombin-activated A subunit of factor XIII (1 U/ml), whereas the iodoacetamide-inactivated A subunit as well as the B subunit had no effect on permeability. Endothelial monolayers exposed to the activated factor XIII A exhibited immunoreactive deposition of itself at interfaces of adjacent cells; however, these were not found on exposure to nonactivated factor XIII A or factor XIII B. Hyperpermeability induced by metabolic inhibition (1 mM potassium cyanide plus 1 mM 2-deoxy-D-glucose) was prevented in the presence of the activated factor XIII A. Likewise, the increase in myocardial water content in ischemic-reperfused rat hearts was prevented in its presence. This study shows that activated factor XIII reduces endothelial permeability. It can prevent the loss of endothelial barrier function under conditions of energy depletion. Its effect seems related to a modification of the paracellular passageways in endothelial monolayers.

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Immunostaining of factor XIII B in endothelial monolayers.  (A) Endothelial cells were incubated for 20 min in the presence of isolated  factor XIII B subunit (10 μg/ml) which had been pretreated with thrombin. Only background fluorescence is observed. (B) Endothelial cells were  exposed to factor XIII B subunit (10 μg/ml) which was not pretreated.  Only background fluorescence is present. (C) Endothelial cells not preincubated with factor XIII B subunit were exposed to anti-factor XIII B  and FITC-coupled anti–rabbit IgG antibody (first and second antibody  control). There is only background fluorescence. Bar, 50 μM.
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Figure 7: Immunostaining of factor XIII B in endothelial monolayers. (A) Endothelial cells were incubated for 20 min in the presence of isolated factor XIII B subunit (10 μg/ml) which had been pretreated with thrombin. Only background fluorescence is observed. (B) Endothelial cells were exposed to factor XIII B subunit (10 μg/ml) which was not pretreated. Only background fluorescence is present. (C) Endothelial cells not preincubated with factor XIII B subunit were exposed to anti-factor XIII B and FITC-coupled anti–rabbit IgG antibody (first and second antibody control). There is only background fluorescence. Bar, 50 μM.

Mentions: In a second set of experiments, endothelial monolayers were incubated in the presence of factor XIII B (10 μg protein/ml) which had been preexposed or not to thrombin. For immunohistochemistry, a specific polyclonal antibody raised against factor XIII B (32) was used, which we confirmed to stain isolated factor XIII B (not shown). No specific staining for factor XIII B was detected in the monolayers (Fig. 7).


Effect of factor XIII on endothelial barrier function.

Noll T, Wozniak G, McCarson K, Hajimohammad A, Metzner HJ, Inserte J, Kummer W, Hehrlein FW, Piper HM - J. Exp. Med. (1999)

Immunostaining of factor XIII B in endothelial monolayers.  (A) Endothelial cells were incubated for 20 min in the presence of isolated  factor XIII B subunit (10 μg/ml) which had been pretreated with thrombin. Only background fluorescence is observed. (B) Endothelial cells were  exposed to factor XIII B subunit (10 μg/ml) which was not pretreated.  Only background fluorescence is present. (C) Endothelial cells not preincubated with factor XIII B subunit were exposed to anti-factor XIII B  and FITC-coupled anti–rabbit IgG antibody (first and second antibody  control). There is only background fluorescence. Bar, 50 μM.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC2193057&req=5

Figure 7: Immunostaining of factor XIII B in endothelial monolayers. (A) Endothelial cells were incubated for 20 min in the presence of isolated factor XIII B subunit (10 μg/ml) which had been pretreated with thrombin. Only background fluorescence is observed. (B) Endothelial cells were exposed to factor XIII B subunit (10 μg/ml) which was not pretreated. Only background fluorescence is present. (C) Endothelial cells not preincubated with factor XIII B subunit were exposed to anti-factor XIII B and FITC-coupled anti–rabbit IgG antibody (first and second antibody control). There is only background fluorescence. Bar, 50 μM.
Mentions: In a second set of experiments, endothelial monolayers were incubated in the presence of factor XIII B (10 μg protein/ml) which had been preexposed or not to thrombin. For immunohistochemistry, a specific polyclonal antibody raised against factor XIII B (32) was used, which we confirmed to stain isolated factor XIII B (not shown). No specific staining for factor XIII B was detected in the monolayers (Fig. 7).

Bottom Line: The thrombin-activated plasma factor XIII (1 U/ml) reduced albumin permeability of endothelial monolayers within 20 min by 30 +/- 7% (basal value of 5.9 +/- 0.4 x 10(-6) cm/s), whereas the nonactivated plasma factor XIII had no effect.This study shows that activated factor XIII reduces endothelial permeability.Its effect seems related to a modification of the paracellular passageways in endothelial monolayers.

View Article: PubMed Central - PubMed

Affiliation: Physiologisches Institut, Justus-Liebig-Universität, D-35392 Giessen, Germany. thomas.noll@physiologie.med.uni-giessen.de

ABSTRACT
The effect of factor XIII on endothelial barrier function was studied in a model of cultured monolayers of porcine aortic endothelial cells and saline-perfused rat hearts. The thrombin-activated plasma factor XIII (1 U/ml) reduced albumin permeability of endothelial monolayers within 20 min by 30 +/- 7% (basal value of 5.9 +/- 0.4 x 10(-6) cm/s), whereas the nonactivated plasma factor XIII had no effect. Reduction of permeability to the same extent, i.e., by 34 +/- 9% could be obtained with the thrombin-activated A subunit of factor XIII (1 U/ml), whereas the iodoacetamide-inactivated A subunit as well as the B subunit had no effect on permeability. Endothelial monolayers exposed to the activated factor XIII A exhibited immunoreactive deposition of itself at interfaces of adjacent cells; however, these were not found on exposure to nonactivated factor XIII A or factor XIII B. Hyperpermeability induced by metabolic inhibition (1 mM potassium cyanide plus 1 mM 2-deoxy-D-glucose) was prevented in the presence of the activated factor XIII A. Likewise, the increase in myocardial water content in ischemic-reperfused rat hearts was prevented in its presence. This study shows that activated factor XIII reduces endothelial permeability. It can prevent the loss of endothelial barrier function under conditions of energy depletion. Its effect seems related to a modification of the paracellular passageways in endothelial monolayers.

Show MeSH
Related in: MedlinePlus