Graefe's archive for clinical and experimental ophthalmology = Albrecht von Graefes Archiv fur klinische und experimentelle Ophthalmologie · 2012
Novel bioactivity of NHERF1 in corneal neovascularization.
СтатьяПоддержка исследования: вне правительства СШАНазвание и аннотация: Перевод пока не подготовлен
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Chen Peng, Wang Ye, Yang Lingling, Li Changyou, Wang Yao, Xie Lixin, Wang Yiqiang
Аннотация
PURPOSE: This paper aimed to determine the involvement of Na(+)/H(+) exchanger regulatory factor 1 (NHERF1) in experimental murine corneal neovascularization (NV), and to study the effect of NHERF1 on the biological properties of HUVEC and related mechanisms.
METHODS: Using loss- and gain-function, we investigated the biological effects of NHERF1 on HUVEC. Western blotting was used to detect the expression of NHERF1 in cells. A carboxyfluorescein succinimidyl ester (CFSE) labeling assay and scarification test were used to measure the proliferation and migration activity, respectively, of HUVEC. The cell cycle distribution of the cells was assessed by flow cytometry analysis. The effect of NHERF1 on the phosphorylation levels of Akt and the changes of matrix metalloproteinase (MMP)-2 and MMP-9 levels were detected by western blotting analysis. Change in the NHERF1 expression in the alkali burn-induced corneal NV model was detected by microarray, real-time PCR, and immunofluorescence.
RESULTS: Overexpression of exogenous NHERF1 in HUVEC slightly inhibited the proliferation and significantly reduced the migration of the cells. NHERF1 also significantly downregulated Akt1 phosphorylation induced by platelet-derived growth factor BB (PDGF-BB) and the secretion of MMP-2 and MMP-9 compared with control cells. NHERF1 was upregulated in corneas challenged with alkali burns.
CONCLUSIONS: Our results indicated that NHERF1 might serve as a potential target for manipulating neovascularization-related diseases. This discovery contributes to a better understanding of the bioactivity of NHERF1 in angiogenesis.
METHODS: Using loss- and gain-function, we investigated the biological effects of NHERF1 on HUVEC. Western blotting was used to detect the expression of NHERF1 in cells. A carboxyfluorescein succinimidyl ester (CFSE) labeling assay and scarification test were used to measure the proliferation and migration activity, respectively, of HUVEC. The cell cycle distribution of the cells was assessed by flow cytometry analysis. The effect of NHERF1 on the phosphorylation levels of Akt and the changes of matrix metalloproteinase (MMP)-2 and MMP-9 levels were detected by western blotting analysis. Change in the NHERF1 expression in the alkali burn-induced corneal NV model was detected by microarray, real-time PCR, and immunofluorescence.
RESULTS: Overexpression of exogenous NHERF1 in HUVEC slightly inhibited the proliferation and significantly reduced the migration of the cells. NHERF1 also significantly downregulated Akt1 phosphorylation induced by platelet-derived growth factor BB (PDGF-BB) and the secretion of MMP-2 and MMP-9 compared with control cells. NHERF1 was upregulated in corneas challenged with alkali burns.
CONCLUSIONS: Our results indicated that NHERF1 might serve as a potential target for manipulating neovascularization-related diseases. This discovery contributes to a better understanding of the bioactivity of NHERF1 in angiogenesis.
Предметные рубрики MeSH
ActinsAnimalsBlotting, WesternBurns, ChemicalCell CycleCell MovementCell ProliferationCorneal NeovascularizationCorneal UlcerEye BurnsFemaleFlow CytometryFluorescent Antibody Technique, IndirectHuman Umbilical Vein Endothelial CellsMatrix Metalloproteinase 2Matrix Metalloproteinase 9MiceMice, Inbred C57BLPhosphoproteinsPhosphorylationPlatelet-Derived Growth FactorProto-Oncogene Proteins c-aktRNA InterferenceReal-Time Polymerase Chain ReactionSodium HydroxideSodium-Hydrogen ExchangersTransfection