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Title: Suppression of retinal neovascularization by shRNA targeting HIF-1alpha. Author: Xia XB, Xiong SQ, Xu HZ, Jiang J, Li Y. Journal: Curr Eye Res; 2008 Oct; 33(10):892-902. PubMed ID: 18853324. Abstract: PURPOSE: To investigate whether vector-based HIF-1alpha -targeted shRNA expression system (pSUPER(siHIF-1alpha)) can inhibit HIF-1alpha and VEGF expression in vitro and suppress retinal neovascularization in the murine model of oxygen-induced retinopathy. METHODS: pSUPER(siHIF-1alpha) from which siRNA targeting HIF-1alpha could be generated was constructed and transfected to human umbilical vein endothelial cell lines (HUVECs). Then the expression levels of HIF-1alpha and VEGF in the cultured cells were measured by RT-PCR, immunoblot, and ELISA assays. Subsequently, pSUPER(siHIF-1alpha)was intravitreally injected into the murine model of oxygen-induced retinopathy (OIR). Retinal neovascularization was evaluated by angiography using fluorescein-labeled dextran and quantitated histologically. Moreover, RT-PCR and immunoblot analysis were used to determine whether local administration of pSUPER(siHIF-1alpha)could affect the expression levels of HIF-1alpha and VEGF in murine retinas. RESULTS: HIF-1alpha and hypoxia-induced vascular endothelial growth factor (VEGF) increase in cultured cells were greatly abolished by pSUPER(siHIF-1alpha). Meanwhile, retinal neovascularization in the eye with pSUPER(siHIF-1alpha)injection was significantly reduced compared with that of the contralateral control eye. Histological analysis indicates that neovascular nuclei protruding into the vitreous cavity were decreased by nearly 65%. Furthermore, HIF-1alpha and VEGF expression levels were down-regulated in the murine retinas treated with pSUPER(siHIF-1alpha). CONCLUSIONS: RNAi targeting HIF-1alpha could inhibit the retinal neovascularization by approximately 65% through down-regulating the expression of HIF-1alpha and VEGF in the murine retinas, which may provide a powerful and novel therapeutic tool for ischemic-induced retinal diseases.[Abstract] [Full Text] [Related] [New Search]