title: Blockade of PKC-beta protects HUVEC from advanced glycation end products induced inflammation.
abstract: Advanced glycation end products (AGEs) have been recognized as a pivotal inducer in diabetes and kinds of aging-related vasculopathy. Endothelial dysfunction and inflammatory cells adhesion to endothelium have been regarded as important and early factors in the pathogenesis of vascular complications in diabetic patients. Owing to the key role of PKC-beta in AGEs-induced vascular dysfunction, we investigated effects of blocking PKC-beta by LY333531 on macrophage adhesion to HUVEC and the related mechanism. Transwell HUVEC-macrophage co-culture system was established to evaluate macrophage migration and adhesion ability. Immunocytochemistry was applied to examine TGF-beta1, ICAM-1 and RAGE protein expressions by SABC or SABC-AP method; mRNA expression of TGF-beta1, ICAM-1 and RAGE was determined by real-time RT-PCR. SOD and MDA levels in culture supernatant were detected. We found that LY333531 significantly reduced AGEs-induced macrophage adhesion to HUVEC. Blockade of PKC-beta strikingly decreased HUVEC TGF-beta1 and ICAM-1 expression in both protein and mRNA levels, RAGE protein level was also down-regulated. Furthermore, the anti-oxidative stress index, SOD/MDA was dramatically elevated on LY333531 application. Therefore we conclude that LY333531 can reduce AGEs-induced macrophage adhesion to endothelial cells and relieve the local inflammation, this was realized by its effect on decreasing inflammatory cytokines' expression and increasing cell anti-oxidative ability.
authors.affiliation: Faculty of Chinese Medicine, Macau University of Science and Technology, Avenida Wai Long, Taipa, Macau.
authors.forename: Youhua
authors.initials: Y
authors.lastname: Xu
authors.forename: Shanshan
authors.initials: S
authors.lastname: Wang
authors.forename: Liang
authors.initials: L
authors.lastname: Feng
authors.forename: Quan
authors.initials: Q
authors.lastname: Zhu
authors.forename: Ping
authors.initials: P
authors.lastname: Xiang
authors.forename: Bao
authors.initials: B
authors.lastname: He
mesh_terms: D000818:Animals; D002448:Cell Adhesion / Q000187:drug effects / Q000276:immunology; D018929:Cell Culture Techniques; D002460:Cell Line; D002465:Cell Movement / Q000187:drug effects / Q000276:immunology; D042783:Endothelial Cells / Q000187:drug effects* / Q000201:enzymology / Q000276:immunology; D004730:Endothelium, Vascular / Q000187:drug effects* / Q000201:enzymology / Q000276:immunology; D017127:Glycation End Products, Advanced / Q000276:immunology* / Q000494:pharmacology; D006801:Humans; D007150:Immunohistochemistry; D007211:Indoles / Q000494:pharmacology; D007249:Inflammation / Q000276:immunology; D008264:Macrophages / Q000187:drug effects / Q000276:immunology; D008297:Male; D008301:Maleimides / Q000494:pharmacology; D008315:Malondialdehyde / Q000378:metabolism; D051379:Mice; D011493:Protein Kinase C / Q000037:antagonists & inhibitors*; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013482:Superoxide Dismutase / Q000378:metabolism
chemical_list: D017127:Glycation End Products, Advanced; D007211:Indoles; D008301:Maleimides; D047428:Protein Kinase Inhibitors; D008315:Malondialdehyde; C099154:ruboxistaurin; D013482:Superoxide Dismutase; D011493:Protein Kinase C