Overproduction of N(epsilon)-(carboxymethyl)lysine-induced neovascularization in cultured choroidal explant of aged rat.
N(epsilon)-(carboxymethyl)lysine (CML) adduct, a major structure of advanced glycation end product, facilitated production of immature microvessels from choroidal explant cultured in fibrin gel. The present study was investigated an action of endogenous CML adduct on neovascularization of cultured choroidal explants of aged Wistar rats with 9 months of age. The number of microvessels budded from explants was counted under optical microscope and used as an index of in vitro neovascularization. Aged choroidal explants increased the neovascularization in an age-dependent manner. Anti-CML antibody decreased age-facilitated neovascularization as well as CML-human serum albumin (HSA)-facilitated neovascularization. Both the aged explant and CML-HSA-treated explant significantly released vascular endothelial growth factor (VEGF), tumor necrosis factor (TNF) alpha and platelet-derived growth factor (PDGF)-B during the culture period. The release of TNF alpha and PDGF-B was earlier than that of VEGF from the aged explants. The antibodies against these factors decreased the CML-facilitated and age-facilitated neovascularization in the choroidal explants. The inhibitory capacity of anti-TNF alpha antibody was greater than those of anti-VEGF and anti-PDGF-B antibodies. In conclusion, endogenous CML adduct overproduced the neovascularization of the aged choroidal explant. The CML adduct releases TNF alpha which might induce the production and release of VEGF for the abnormal choroidal neovascularization in the patients of age-related macular degeneration.
Kobayashi
Shinjiro
S
Department of Clinical Pharmacology, Faculty of Pharmaceutical Sciences, Hokuriku University, Kanazawa, Japan. s-kobayashi@hokuriku-u.ac.jp
Nomura
Masaaki
M
Nishioka
Tatsuo
T
Kikuchi
Minoru
M
Ishihara
Akina
A
Nagai
Ryoji
R
Hagino
Nobuyoshi
N
D000367:Age Factors; D000375:Aging / Q000378:metabolism*; D000818:Animals; D000906:Antibodies / Q000494:pharmacology; D002196:Capillaries / Q000187:drug effects / Q000378:metabolism*; D002829:Choroid / Q000098:blood supply*; D020256:Choroidal Neovascularization / Q000378:metabolism*; D017127:Glycation End Products, Advanced / Q000276:immunology / Q000378:metabolism*; D008239:Lysine / Q000031:analogs & derivatives* / Q000276:immunology / Q000378:metabolism; D008268:Macular Degeneration / Q000378:metabolism / Q000503:physiopathology; D008297:Male; D009924:Organ Culture Techniques; D010982:Platelet-Derived Growth Factor / Q000276:immunology / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D013997:Time Factors; D014409:Tumor Necrosis Factor-alpha / Q000276:immunology / Q000378:metabolism; D042461:Vascular Endothelial Growth Factor A / Q000276:immunology / Q000378:metabolism
D000906:Antibodies; D017127:Glycation End Products, Advanced; D010982:Platelet-Derived Growth Factor; D014409:Tumor Necrosis Factor-alpha; D042461:Vascular Endothelial Growth Factor A; C467498:vascular endothelial growth factor A, rat; C048496:N(6)-carboxymethyllysine; D008239:Lysine