[Effects of Rho/ROCK signal pathway on AGEs-induced morphological and functional changes in human dermal microvascular endothelial cells.].
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.
Department of Pathophysiology, Southern Medical University, Guangzhou, China.
Ji-Ping
JP
Wang
Xiao-Hua
XH
Guo
Ling-Jun
LJ
Wang
Qiang
Q
Li
Bo
B
Chen
Wei
W
Wu
Xu-Liang
XL
Huang
Qiao-Bing
QB
Huang
D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine / Q000031:analogs & derivatives / Q000494:pharmacology; D008841:Actin Cytoskeleton / Q000378:metabolism; D000199:Actins / Q000378:metabolism; D000577:Amides / Q000494:pharmacology; D042783:Endothelial Cells / Q000378:metabolism*; D004730:Endothelium, Vascular / Q000166:cytology; D016650:Fluorescein-5-isothiocyanate / Q000031:analogs & derivatives / Q000378:metabolism; D017127:Glycation End Products, Advanced / Q000494:pharmacology*; D006801:Humans; D010590:Phalloidine / Q000031:analogs & derivatives; D010766:Phosphorylation; D011725:Pyridines / Q000494:pharmacology; D012235:Rhodamines; D012709:Serum Albumin / Q000378:metabolism / Q000494:pharmacology*; D000075462:Serum Albumin, Human; D015398:Signal Transduction; D054460:rho-Associated Kinases / Q000378:metabolism*
C459092:2-methyl-1-((4-methyl-5-isoquinolinyl)sulfonyl)homopiperazine; D000199:Actins; D000577:Amides; C518288:FITC-albumin; D017127:Glycation End Products, Advanced; D011725:Pyridines; D012235:Rhodamines; D012709:Serum Albumin; C109748:advanced glycation end products-human serum albumin; C504731:rhodamine-phalloidin; C108830:Y 27632; D010590:Phalloidine; D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; D054460:rho-Associated Kinases; D016650:Fluorescein-5-isothiocyanate; D000075462:Serum Albumin, Human