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Dexamethasone Suppressed LPS-Induced Matrix Metalloproteinase and Its Effect on Endothelial Glycocalyx Shedding.

Cui N, Wang H, Long Y, Su L, Liu D - Mediators Inflamm. (2015)

Bottom Line: The aim of this study is to determine the mechanism of sepsis-induced vascular hyperpermeability and the beneficial effect of glucocorticoid in protecting vascular endothelium.The inhibition of MMPs by dexamethasone was significantly lower than that by doxycycline, while the rescue of syndecan-1 expression from LPS-induced endotoxemic rat thoracic aorta was significantly higher in the dexamethasone-treated compared to the doxycycline-treated (p = 0.03).In conclusion, activation of MMPs plays important role in regulating ZO-1 and syndecan-1 protein levels in LPS mediated endothelial perturbation.

View Article: PubMed Central - PubMed

Affiliation: Department of Critical Care Medicine, Peking Union Medical College Hospital, Beijing 100730, China.

ABSTRACT
The aim of this study is to determine the mechanism of sepsis-induced vascular hyperpermeability and the beneficial effect of glucocorticoid in protecting vascular endothelium. Male Sprague-Dawley rats were given either a bolus intraperitoneal injection of a nonlethal dose of LPS (Escherichia coli 055:B5, 10 mg/kg, Sigma) or vehicle (pyrogen-free water). Animals of treatment groups were also given either dexamethasone (4 mg/kg, 30 min prior to LPS injection) or the matrix metalloproteinases (MMPs) inhibitor doxycycline (4 mg/kg, 30 min after LPS injection). Both activities and protein levels of MMP-2 (p < 0.001) and MMP-9 (p < 0.001) were significantly upregulated in aortic homogenates from LPS-treated rats, associated with decreased ZO-1 (p < 0.001) and syndecan-1 (p = 0.011) protein contents. Both dexamethasone and doxycycline could significantly inhibit MMPs activity and reserve the expressions of ZO-1 and syndecan-1. The inhibition of MMPs by dexamethasone was significantly lower than that by doxycycline, while the rescue of syndecan-1 expression from LPS-induced endotoxemic rat thoracic aorta was significantly higher in the dexamethasone-treated compared to the doxycycline-treated (p = 0.03). In conclusion, activation of MMPs plays important role in regulating ZO-1 and syndecan-1 protein levels in LPS mediated endothelial perturbation. Both dexamethasone and doxycycline inhibit activation of MMPs that may contribute to the rescue of ZO-1 and syndecan-1 expression.

No MeSH data available.


Related in: MedlinePlus

Gelatin zymography. Proteins of the aortic homogenate from each group were used for zymographic assay as described in the method. (a) One representative image data of zymogram. 72-kDa: latent MMP-2; 82-kDa: active MMP-9; and 92-kDa: latent MMP-9. (b, c, and d) Densitometric analysis of 8 biological replicates of latent MMP-2 (72-kDa), active MMP-9 (82-kDa), and latent MMP-9 (92-kDa), respectively. ∗p < 0.05; ∗∗p < 0.001.
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fig2: Gelatin zymography. Proteins of the aortic homogenate from each group were used for zymographic assay as described in the method. (a) One representative image data of zymogram. 72-kDa: latent MMP-2; 82-kDa: active MMP-9; and 92-kDa: latent MMP-9. (b, c, and d) Densitometric analysis of 8 biological replicates of latent MMP-2 (72-kDa), active MMP-9 (82-kDa), and latent MMP-9 (92-kDa), respectively. ∗p < 0.05; ∗∗p < 0.001.

Mentions: In order to determine the effect of dexamethasone or doxycycline on MMP expression and activation, a gelatin zymogram was performed. To accomplish this, equal amount of protein from each group aortic tissue homogenate was loaded in a 10% PAGE-SDS gel with gelatin as substrate. As shown in Figure 2, under “Control” condition or in the presence of dexamethasone alone, neither MMP-2 nor MMP-9 was detected by gelatin zymogram. In contrast, both MMP-2 and MMP-9 were significantly induced by LPS. Interestingly, however, only MMP-9 but not MMP-2 was activated by LPS in the current study (Figure 2(a)). In the presence of dexamethasone, LPS-induced MMP-9 (both latent and active) and MMP-2 (latent form) were significantly suppressed (Figures 2(b), 2(c), and 2(d), p < 0.05). Similarly, doxycycline not only almost completely blocked latent MMP-2 and MMP-9 production, but also significantly inhibited MMP-9 conversion from latent to active form (Figures 2(b), 2(c), and 2(d), p < 0.05). Consistent with results of gelatin zymogram, immunoblotting of MMP-2 and MMP-9 confirmed that both dexamethasone and doxycycline could significantly suppress LPS-induced MMP-2 or MMP-9 expression in the aortic tissues (Figures 3(a) and 3(b)).


Dexamethasone Suppressed LPS-Induced Matrix Metalloproteinase and Its Effect on Endothelial Glycocalyx Shedding.

Cui N, Wang H, Long Y, Su L, Liu D - Mediators Inflamm. (2015)

Gelatin zymography. Proteins of the aortic homogenate from each group were used for zymographic assay as described in the method. (a) One representative image data of zymogram. 72-kDa: latent MMP-2; 82-kDa: active MMP-9; and 92-kDa: latent MMP-9. (b, c, and d) Densitometric analysis of 8 biological replicates of latent MMP-2 (72-kDa), active MMP-9 (82-kDa), and latent MMP-9 (92-kDa), respectively. ∗p < 0.05; ∗∗p < 0.001.
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4493300&req=5

fig2: Gelatin zymography. Proteins of the aortic homogenate from each group were used for zymographic assay as described in the method. (a) One representative image data of zymogram. 72-kDa: latent MMP-2; 82-kDa: active MMP-9; and 92-kDa: latent MMP-9. (b, c, and d) Densitometric analysis of 8 biological replicates of latent MMP-2 (72-kDa), active MMP-9 (82-kDa), and latent MMP-9 (92-kDa), respectively. ∗p < 0.05; ∗∗p < 0.001.
Mentions: In order to determine the effect of dexamethasone or doxycycline on MMP expression and activation, a gelatin zymogram was performed. To accomplish this, equal amount of protein from each group aortic tissue homogenate was loaded in a 10% PAGE-SDS gel with gelatin as substrate. As shown in Figure 2, under “Control” condition or in the presence of dexamethasone alone, neither MMP-2 nor MMP-9 was detected by gelatin zymogram. In contrast, both MMP-2 and MMP-9 were significantly induced by LPS. Interestingly, however, only MMP-9 but not MMP-2 was activated by LPS in the current study (Figure 2(a)). In the presence of dexamethasone, LPS-induced MMP-9 (both latent and active) and MMP-2 (latent form) were significantly suppressed (Figures 2(b), 2(c), and 2(d), p < 0.05). Similarly, doxycycline not only almost completely blocked latent MMP-2 and MMP-9 production, but also significantly inhibited MMP-9 conversion from latent to active form (Figures 2(b), 2(c), and 2(d), p < 0.05). Consistent with results of gelatin zymogram, immunoblotting of MMP-2 and MMP-9 confirmed that both dexamethasone and doxycycline could significantly suppress LPS-induced MMP-2 or MMP-9 expression in the aortic tissues (Figures 3(a) and 3(b)).

Bottom Line: The aim of this study is to determine the mechanism of sepsis-induced vascular hyperpermeability and the beneficial effect of glucocorticoid in protecting vascular endothelium.The inhibition of MMPs by dexamethasone was significantly lower than that by doxycycline, while the rescue of syndecan-1 expression from LPS-induced endotoxemic rat thoracic aorta was significantly higher in the dexamethasone-treated compared to the doxycycline-treated (p = 0.03).In conclusion, activation of MMPs plays important role in regulating ZO-1 and syndecan-1 protein levels in LPS mediated endothelial perturbation.

View Article: PubMed Central - PubMed

Affiliation: Department of Critical Care Medicine, Peking Union Medical College Hospital, Beijing 100730, China.

ABSTRACT
The aim of this study is to determine the mechanism of sepsis-induced vascular hyperpermeability and the beneficial effect of glucocorticoid in protecting vascular endothelium. Male Sprague-Dawley rats were given either a bolus intraperitoneal injection of a nonlethal dose of LPS (Escherichia coli 055:B5, 10 mg/kg, Sigma) or vehicle (pyrogen-free water). Animals of treatment groups were also given either dexamethasone (4 mg/kg, 30 min prior to LPS injection) or the matrix metalloproteinases (MMPs) inhibitor doxycycline (4 mg/kg, 30 min after LPS injection). Both activities and protein levels of MMP-2 (p < 0.001) and MMP-9 (p < 0.001) were significantly upregulated in aortic homogenates from LPS-treated rats, associated with decreased ZO-1 (p < 0.001) and syndecan-1 (p = 0.011) protein contents. Both dexamethasone and doxycycline could significantly inhibit MMPs activity and reserve the expressions of ZO-1 and syndecan-1. The inhibition of MMPs by dexamethasone was significantly lower than that by doxycycline, while the rescue of syndecan-1 expression from LPS-induced endotoxemic rat thoracic aorta was significantly higher in the dexamethasone-treated compared to the doxycycline-treated (p = 0.03). In conclusion, activation of MMPs plays important role in regulating ZO-1 and syndecan-1 protein levels in LPS mediated endothelial perturbation. Both dexamethasone and doxycycline inhibit activation of MMPs that may contribute to the rescue of ZO-1 and syndecan-1 expression.

No MeSH data available.


Related in: MedlinePlus