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

• Published in: Cardiovascular diabetology Vol. 11; no. 1; p. 65
• Main Authors: Li, Hailing, Peng, Wenhui, Jian, Weixia, Li, Yuanmin, Li, Qi, Li, Weiming, Xu, Yawei
• Format: Journal Article
• Language: English
• Published: England BioMed Central 13.06.2012; BMC
• Subjects: 1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine - administration & dosage; 1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine - analogs & derivatives; 1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine - pharmacology; Aged; Anti-Inflammatory Agents - administration & dosage; Anti-Inflammatory Agents - pharmacology; Blotting, Western; Cardiovascular disease; Cell adhesion & migration; Cell Adhesion - drug effects; Cells, Cultured; Chemokine CCL2 - blood; Chemokine CCL2 - genetics; Chemokine CCL2 - metabolism; Chemotaxis, Leukocyte - drug effects; China; Coculture Techniques; Diabetes Mellitus - blood; Diabetes Mellitus - drug therapy; Diabetes Mellitus - enzymology; Diabetes Mellitus - immunology; Dose-Response Relationship, Drug; Enzyme-Linked Immunosorbent Assay; Fasudil; Female; Gene Expression Regulation - drug effects; Glucose - metabolism; High glucose; Human Umbilical Vein Endothelial Cells - drug effects; Human Umbilical Vein Endothelial Cells - enzymology; Human Umbilical Vein Endothelial Cells - immunology; Humans; Infusions, Intravenous; Male; Middle Aged; Monocyte chemoattractant protein-1; Monocytes - drug effects; Monocytes - immunology; Nitric oxide; Original Investigation; Protein Kinase Inhibitors - administration & dosage; Protein Kinase Inhibitors - pharmacology; Proteins; Real-Time Polymerase Chain Reaction; rho-Associated Kinases - antagonists & inhibitors; rho-Associated Kinases - metabolism; RNA, Messenger - metabolism; Single-Blind Method; Time Factors; Vascular cell adhesion molecule-1; Vascular Cell Adhesion Molecule-1 - blood; Vascular Cell Adhesion Molecule-1 - genetics; Vascular Cell Adhesion Molecule-1 - metabolism
• ISSN: 1475-2840; 1475-2840
• DOI: 10.1186/1475-2840-11-65

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). HUVECs 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. Fasudil 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). Treatment 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.