Effects of Rho/ROCK signal pathway on AGEs-induced morphological and functional changes in human dermal microvascular endothelial cells..
- Author:
Ji-Ping WANG
1
;
Xiao-Hua GUO
;
Ling-Jun WANG
;
Qiang LI
;
Bo CHEN
;
Wei WU
;
Xu-Liang HUANG
;
Qiao-Bing HUANG
Author Information
1. Department of Pathophysiology, Southern Medical University, Guangzhou, China.
- Publication Type:Journal Article
- MeSH:
1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine;
analogs & derivatives;
pharmacology;
Actin Cytoskeleton;
metabolism;
Actins;
metabolism;
Amides;
pharmacology;
Endothelial Cells;
metabolism;
Endothelium, Vascular;
cytology;
Fluorescein-5-isothiocyanate;
analogs & derivatives;
metabolism;
Glycation End Products, Advanced;
pharmacology;
Humans;
Phalloidine;
analogs & derivatives;
Phosphorylation;
Pyridines;
pharmacology;
Rhodamines;
Serum Albumin;
metabolism;
pharmacology;
Serum Albumin, Human;
Signal Transduction;
rho-Associated Kinases;
metabolism
- From:
Acta Physiologica Sinica
2009;61(2):132-138
- CountryChina
- Language:Chinese
-
Abstract:
The present study aimed to determine the role of Rho/Rho kinase (Rho/ROCK) phosphorylation on advanced glycation end products (AGEs)-induced morphological and functional changes in human dermal microvascular endothelial cells (HMVECs). HMVECs were respectively incubated with different concentrations of AGEs-modified human serum albumin (AGEs-HSA) for different time. In some other cases, HMVECs were pretreated with ROCK inhibitors (H-1152 or Y-27632). The morphological changes of F-actin cytoskeleton were visualized by rhodamine-phalloidin staining and the phosphorylation of Rho and ROCK were determined by Western blot. Endothelial monolayer permeability was assessed by measuring the flux of FITC-albumin across the endothelial cells. The results showed that the distribution of F-actin was significantly altered by AGEs-HSA in time and dose-dependent patterns. These effects were inhibited by ROCK inhibitors. The phosphorylation of Rho and RCOK was remarkably increased by AGEs-HSA treatment while total Rho and ROCK protein levels were not affected. The permeability of endothelial monolayer was dramatically increased by AGEs-HSA, and both ROCK inhibitors (H-1152 or Y-27632) attenuated these hyperpermeability responses. The results obtained suggest that the phosphorylation of Rho/ROCK plays an important role in AGEs-induced morphological and functional alterations in HMVECs.
