|Semaphorin 3A blocks the formation of pathologic choroidal neovascularization induced by transforming growth factor beta|
|Bai, Yujing; Liang, Shuting; Yu, Wenzhen; Zhao, Min; Huang, Lvzhen; Zhao, Mingwei; Li, Xiaoxin|
|WOS标题词||Science & Technology|
|类目[WOS]||Biochemistry & Molecular Biology ; Ophthalmology|
|研究领域[WOS]||Biochemistry & Molecular Biology ; Ophthalmology|
|关键词[WOS]||RECEPTOR SIGNALING PATHWAYS ; PIGMENT EPITHELIAL-CELLS ; MACULAR DEGENERATION ; TGF-BETA ; RETINAL NEOVASCULARIZATION ; ANGIOGENESIS ; EXPRESSION ; THERAPY ; NEUROPILINS ; INHIBITION|
Objective: Choroidal neovascularization (CNV) is a major cause of vision loss in retinal diseases such as age-related macular degeneration (AMD). Previously, we demonstrated that semaphorin3A (Sema3A), which is a chemorepellent guidance molecule, inhibited the formation of retina neovascularization. In the present study, we investigated the anti-angiogenic effects of Sema3A on transforming growth factor beta (TGF-beta) in vitro and in vivo.
Methods: Enzyme-linked immunosorbent assays (ELISAs) were used to measure the TGF-beta levels in the vitreous humor of patients with AMD and controls. Human umbilical vein endothelial cells (HUVECs) were used for the in vitro study, and a laser-induced CNV mouse model was prepared for the in vivo study. The HUVECs were incubated with TGF-beta and Sema3A. The proliferation, migration, apoptosis, and tube formation of the cells were then measured using BrdU, Transwell, flow cytometry, and Matrigel assays, respectively, and the SMAD2/3 signaling pathways were analyzed using western blot analysis. The C57BL/6J mouse retina was exposed to a laser to induce choroidal neovascularization (CNV), and Sema3A was injected intravitreously. After 14 days, fundus fluorescein angiography was performed to evaluate the leakage area of the CNV. The vascular endothelial growth factor (VEGF) and TGF-beta concentrations in the retina-choroid complex were measured with ELISA. Components of the p38 mitogen-activated protein kinase (MAPK), extracellular signal-regulated kinase-1/2 (ERK1/2), c-Jun NH2-terminal kinase (JNK), and SMAD2/3 signaling pathways in the Sema3A-treated groups were analyzed using western blotting.
Results: In this study, we first verified that the vitreous TGF-beta level was higher in patients with neovascular AMD than in the controls. We also showed that Sema3A inhibited TGF-beta-induced HUVEC proliferation, migration, and tube formation and inhibited the downstream SMAD2/3 signaling pathway. Sema3A also induced TGF-beta-stimulated HUVEC apoptosis and inhibited the response of TGF-beta in vitro. In vivo, the TGF-beta level was increased in the CNV mouse model. Sema3A not only inhibited laser-induced CNV formation but also inhibited the uptake of VEGF and TGF-beta. In the western blot analysis, Sema3A was shown to inhibit the phosphorylation of p38 MAPK, ERK1/2, and JNK and to inhibit the SMAD2/3 signaling pathway after Sema3A treatment in CNV mice.
Conclusions: Sema3A can be applied as a useful, adjunctive therapeutic strategy for preventing CNV formation.
|项目编号||2011CB510200 ; Z131102000413004 ; 81200690 ; RDB201320 ; RDB201224|
|资助机构||National Basic Research Program of China (973 Program) ; Beijing Nova Program ; National Natural Science Foundation of China Grant ; Peking University People&prime ; s Hospital Research and Development Fund|
|作者单位||Peking Univ, Peoples Hosp, Dept Ophthalmol,Key Lab Vis Loss & Restorat,Minis, Key Lab Diag & Treatment Retinal & Choroid Dis, Beijing 100044, Peoples R China|
|Bai, Yujing,Liang, Shuting,Yu, Wenzhen,et al. Semaphorin 3A blocks the formation of pathologic choroidal neovascularization induced by transforming growth factor beta[J]. MOLECULAR VISION,2014,20:1258-1270.|
|APA||Bai, Yujing.,Liang, Shuting.,Yu, Wenzhen.,Zhao, Min.,Huang, Lvzhen.,...&Li, Xiaoxin.(2014).Semaphorin 3A blocks the formation of pathologic choroidal neovascularization induced by transforming growth factor beta.MOLECULAR VISION,20,1258-1270.|
|MLA||Bai, Yujing,et al."Semaphorin 3A blocks the formation of pathologic choroidal neovascularization induced by transforming growth factor beta".MOLECULAR VISION 20(2014):1258-1270.|