ROCK inhibitor fasudil attenuated high glucose-induced MCP-1 and VCAM-1 expression and monocyte-endothelial cell adhesion

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.

Longistatin in tick saliva blocks advanced glycation end-product receptor activation

“…A wealth of literature suggests that RAGE/ ligand interaction induces upregulation of adhesion molecules (29,30). Here, we also observed that stimulation of HUVECs with Gla-BSA sharply increased VCAM1, ICAM1, and E-selectin mRNA, which was efficiently downregulated by anti-RAGE, indicating that Gla-BSA upregulated the expression of adhesion molecules via RAGE.…”

“…These adhesion molecules play essential roles during the recruitment of inflammatory cells into the site of insults (29,30). We provide data supporting that rlongistatin prevents the secretion of cytokines from endothelial cells induced by RAGE ligands.…”

“…Among the adhesion molecules, robust upregulation of VCAM1 was detected following the RAGE ligand stimulation, which was in turn abruptly attenuated by longistatin. High expression of adhesion molecules in endothelial cells promotes interaction of circulating leukocytes with the endothelial surface, and this can eventually result in pavementation, margination, and extravasations of the reactive cells into the site of tissue injury (30). Furthermore, we also studied the effects of longistatin on the Gla-BSA-induced secretion of cytokines by HUVECs.…”

Irradiated U937 Cells Trigger Inflammatory Bystander Responses in Human Umbilical Vein Endothelial Cells through the p38 Pathway

“…The monocytes can pass through the injured or dysfunctional endothelium into the extravascular space and then differentiate into macrophages (12). These processes are partly regulated by chemotactic factors, such as endothelial VCAM-1, which directly takes part in the development of atherosclerosis as an inflammatory factor (66). The induction of adhesion molecules in vascular endothelial cells and the subsequent recruitment of circulating monocytes are proinflammatory events that promote atherogenesis and plaque instability (67).…”

Vascular Tissue Contractility Changes Following Late Gestational Exposure to Multi-Walled Carbon Nanotubes or their Dispersing Vehicle in Sprague Dawley Rats

“…The active form of RhoA promotes activation of the Rho kinase (ROCK) that inhibits MLC phosphatase (MLCP) activity, the dephosphorylation of myosin and subsequent relaxation [24]. Alterations in the RhoA/Rho kinase pathway is reported to be involved in endothelial dysfunction, inflammation [25,26] and with exposure to particulate matter [27]. …”