Abstract
BackgroundPrevious studies suggested that the RhoA/ROCK pathway may contribute to vascular complications in diabetes. The present study was designed to investigate whether ROCK inhibitor fasudil could prevent high glucose-induced monocyte-endothelial cells adhesion, and whether this was related to fasudil effects on vascular endothelial cell expression of chemotactic factors, vascular cell adhesion molecule-1 (VCAM-1) and monocyte chemoattractant protein-1 (MCP-1).MethodsHUVECs were stimulated with high glucose (HG) or HG + fasudil in different concentration or different time. Monocyte-endothelial cell adhesion was determined using fluorescence-labeled monocytes. The mRNA and protein expression of VCAM-1 and MCP-1 were measured using real-time PCR and western blot. The protein levels of RhoA, ROCKI and p-MYPT were determined using western blot analysis. ELISA was employed to measure the expression of soluble VCAM-1 and MCP-1 in cell supernatants and human serum samples.ResultsFasudil significantly suppressed HG-induced adhesion of THP-1 to HUVECs. Fasudil reduced Rho/ROCK activity (as indicated by lower p-MYPT/MYPT ratio), and prevented HG induced increases in VCAM-1 and MCP-1 mRNA and protein levels. Fasudil also decreased MCP-1 concentration in HUVEC supernatants, but increased sVCAM-1 shedding into the media. In human diabetic subjects, 2 weeks of fasudil treatment significantly decreased serum MCP-1 level from 27.9 ± 10.6 pg/ml to 13.8 ± 7.0 pg/ml (P < 0.05), while sVCAM-1 increased from 23.2 ± 7.5 ng/ml to 39.7 ± 5.6 ng/ml after fasudil treatment (P < 0.05).ConclusionsTreatment with the Rho/ROCK pathway inhibitor fasudil attenuated HG-induced monocyte-endothelial cell adhesion, possibly by reducing endothelial expression of VCAM-1 and MCP-1. These results suggest inhibition of Rho/ROCK signaling may have therapeutic potential in preventing diabetes associated vascular inflammation and atherogenesis.
Highlights
Macrovascular complications including atherosclerosis are the leading causes of morbidity and mortality in patients with diabetes mellitus [1]
Fasudil inhibited the high glucose (HG)-mediated monocyte-endothelial cells adhesion in vitro We used BCECF-AM labeled monocytes to evaluate the effect of fasudil on adhesion of monocytes to Human umbilical vein endothelial cells (HUVECs)
The present study demonstrated that Rho kinase (ROCK) inhibitor fasudil attenuated HG-induced increases in vascular cell adhesion molecule-1 (VCAM-1) and monocyte chemoattractant protein-1 (MCP-1) expression in HUVECs
Summary
Macrovascular complications including atherosclerosis are the leading causes of morbidity and mortality in patients with diabetes mellitus [1]. Transmigration of monocytes into the subendothelial space and subsequent transformation into macrophagederived foam cells are key events in atherogenesis [2,3] These processes are partly regulated by chemotactic factors such as monocyte chemoattractant protein-1 (MCP-1) and endothelial vascular cell adhesion molecule-1 (VCAM-1) [4,5]. Recent studies suggest that the RhoA/ROCK pathway may contribute to diabetic vascular complications [6,7,8]. Previous studies suggested that the RhoA/ROCK pathway may contribute to vascular complications in diabetes. The present study was designed to investigate whether ROCK inhibitor fasudil could prevent high glucose-induced monocyte-endothelial cells adhesion, and whether this was related to fasudil effects on vascular endothelial cell expression of chemotactic factors, vascular cell adhesion molecule-1 (VCAM-1) and monocyte chemoattractant protein-1 (MCP-1)
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