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Role of Src in Vascular Hyperpermeability Induced by Advanced Glycation End Products.

Zhang W, Xu Q, Wu J, Zhou X, Weng J, Xu J, Wang W, Huang Q, Guo X - Sci Rep (2015)

Bottom Line: Activation of Src with pcDNA3/flag-Src(Y530F) alone duplicated these effects.Inhibition of Src with siRNA, PP2 or pcDNA3/flag-Src(K298M) abolished these effects.Up-regulation of Src activity induced the phosphorylation of moesin, as well as activation and dissociation of VE-cadherin, while down-regulation of Src abolished these effects.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathophysiology, Key Laboratory for Shock and Microcirculation Research of Guangdong Province, Southern Medical University, Guangzhou 510515, China.

ABSTRACT
The disruption of microvascular barrier in response to advanced glycation end products (AGEs) stimulation contributes to vasculopathy associated with diabetes mellitus. Here, to study the role of Src and its association with moesin, VE-cadherin and focal adhesion kinase (FAK) in AGE-induced vascular hyperpermeability, we verified that AGE induced phosphorylation of Src, causing increased permeability in HUVECs. Cells over-expressed Src displayed a higher permeability after AGE treatment, accompanied with more obvious F-actin rearrangement. Activation of Src with pcDNA3/flag-Src(Y530F) alone duplicated these effects. Inhibition of Src with siRNA, PP2 or pcDNA3/flag-Src(K298M) abolished these effects. The pulmonary microvascular endothelial cells (PMVECs) isolated from receptor for AGEs (RAGE)-knockout mice decreased the phosphorylation of Src and attenuated the barrier dysfunction after AGE-treatment. In vivo study showed that the exudation of dextran from mesenteric venules was increased in AGE-treated mouse. This was attenuated in RAGE knockout or PP2-pretreated mice. Up-regulation of Src activity induced the phosphorylation of moesin, as well as activation and dissociation of VE-cadherin, while down-regulation of Src abolished these effects. FAK was also proved to interact with Src in HUVECs stimulated with AGEs. Our studies demonstrated that Src plays a critical role in AGE-induced microvascular hyperpermeability by phosphorylating moesin, VE-cadherin, and FAK respectively.

No MeSH data available.


Related in: MedlinePlus

PP2 and knockout of RAGE prevents AGE-induced microvascular hyperpermeability.(a,b) Wild type (WT) or RAGE−/− mice were pretreated with AGEs or AGEs plus PP2, The exudation of FITC-dextran from mesenteric venules was determined. Vascular permeability is expressed as the relative fluorescent intensity inside the vessel to that outside the vessel. n = 3, *P < 0.05 versus control group, #P < 0.05 versus AGEs. Each image was recorded at a 100 × magnification.
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f5: PP2 and knockout of RAGE prevents AGE-induced microvascular hyperpermeability.(a,b) Wild type (WT) or RAGE−/− mice were pretreated with AGEs or AGEs plus PP2, The exudation of FITC-dextran from mesenteric venules was determined. Vascular permeability is expressed as the relative fluorescent intensity inside the vessel to that outside the vessel. n = 3, *P < 0.05 versus control group, #P < 0.05 versus AGEs. Each image was recorded at a 100 × magnification.

Mentions: The role of Src and RAGE in microvascular permeability was investigated in AGE-treated wild type (WT) and RAGE knockout mice. We revealed no exudation of FITC-dextran out of the venules in WT mice without AGE treatment. AGE-treated WT mice showed a significant increase in the exudation of FITC-dextran in the extravascular space. This exudation was markedly reduced by a pan Src inhibitor, PP2, and completely abated in RAGE knockout mice (Fig. 5). These findings suggest that Src and RAGE played an important role in AGE-induced microvascular hyperpermeability.


Role of Src in Vascular Hyperpermeability Induced by Advanced Glycation End Products.

Zhang W, Xu Q, Wu J, Zhou X, Weng J, Xu J, Wang W, Huang Q, Guo X - Sci Rep (2015)

PP2 and knockout of RAGE prevents AGE-induced microvascular hyperpermeability.(a,b) Wild type (WT) or RAGE−/− mice were pretreated with AGEs or AGEs plus PP2, The exudation of FITC-dextran from mesenteric venules was determined. Vascular permeability is expressed as the relative fluorescent intensity inside the vessel to that outside the vessel. n = 3, *P < 0.05 versus control group, #P < 0.05 versus AGEs. Each image was recorded at a 100 × magnification.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4585381&req=5

f5: PP2 and knockout of RAGE prevents AGE-induced microvascular hyperpermeability.(a,b) Wild type (WT) or RAGE−/− mice were pretreated with AGEs or AGEs plus PP2, The exudation of FITC-dextran from mesenteric venules was determined. Vascular permeability is expressed as the relative fluorescent intensity inside the vessel to that outside the vessel. n = 3, *P < 0.05 versus control group, #P < 0.05 versus AGEs. Each image was recorded at a 100 × magnification.
Mentions: The role of Src and RAGE in microvascular permeability was investigated in AGE-treated wild type (WT) and RAGE knockout mice. We revealed no exudation of FITC-dextran out of the venules in WT mice without AGE treatment. AGE-treated WT mice showed a significant increase in the exudation of FITC-dextran in the extravascular space. This exudation was markedly reduced by a pan Src inhibitor, PP2, and completely abated in RAGE knockout mice (Fig. 5). These findings suggest that Src and RAGE played an important role in AGE-induced microvascular hyperpermeability.

Bottom Line: Activation of Src with pcDNA3/flag-Src(Y530F) alone duplicated these effects.Inhibition of Src with siRNA, PP2 or pcDNA3/flag-Src(K298M) abolished these effects.Up-regulation of Src activity induced the phosphorylation of moesin, as well as activation and dissociation of VE-cadherin, while down-regulation of Src abolished these effects.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathophysiology, Key Laboratory for Shock and Microcirculation Research of Guangdong Province, Southern Medical University, Guangzhou 510515, China.

ABSTRACT
The disruption of microvascular barrier in response to advanced glycation end products (AGEs) stimulation contributes to vasculopathy associated with diabetes mellitus. Here, to study the role of Src and its association with moesin, VE-cadherin and focal adhesion kinase (FAK) in AGE-induced vascular hyperpermeability, we verified that AGE induced phosphorylation of Src, causing increased permeability in HUVECs. Cells over-expressed Src displayed a higher permeability after AGE treatment, accompanied with more obvious F-actin rearrangement. Activation of Src with pcDNA3/flag-Src(Y530F) alone duplicated these effects. Inhibition of Src with siRNA, PP2 or pcDNA3/flag-Src(K298M) abolished these effects. The pulmonary microvascular endothelial cells (PMVECs) isolated from receptor for AGEs (RAGE)-knockout mice decreased the phosphorylation of Src and attenuated the barrier dysfunction after AGE-treatment. In vivo study showed that the exudation of dextran from mesenteric venules was increased in AGE-treated mouse. This was attenuated in RAGE knockout or PP2-pretreated mice. Up-regulation of Src activity induced the phosphorylation of moesin, as well as activation and dissociation of VE-cadherin, while down-regulation of Src abolished these effects. FAK was also proved to interact with Src in HUVECs stimulated with AGEs. Our studies demonstrated that Src plays a critical role in AGE-induced microvascular hyperpermeability by phosphorylating moesin, VE-cadherin, and FAK respectively.

No MeSH data available.


Related in: MedlinePlus