Effect And Mechanisms Of BV8 On Retinal Neovascularization In Mice

Purpose: Retinal neovascularization(RNV)is one of the main causes of irreversible retinopathy and visual impairment.Its pathological process is complicated and it is the result of the interaction of various cells,pro-angiogenic factors and inflammatory factors.BV8(Bombina variegate 8)is also known as Prokineticins-2(PK-2).Various studies have confirmed that BV8 is a potent pro-angiogenic factor in various tissues and organs.Therefore,this study used two mouse RNV disease models to explore the role of BV8 in the process of RNV and its related mechanisms.Methods: This subject is mainly divided into two parts,the first part is the animal experiment in vivo.BV8 expression in Oxygen-induced Retinopathy(OIR)mice model and Rhodopsin/VEGF(Rho/VEGF)transgenic mice model was detected by Real Time-PCR,Western Blot and immunofluorescence staining.With intravitreal injection of anti-BV8 neutralizing antibodies in two mouse models,retinal flat-mounts and molecular biology methods were performed to detect the effects of BV8 on RNV formation and pro-angiogenic factors.The second part is the cell experiment in vitro.Immunofluorescence staining and molecular biological methods were used to observe the expression of BV8 cells in human Retinal Microvascular Endothelial Cells(HRECs)cells in hypoxia.The tube formation assay and Cell counting kit-8 proliferation assay were used to evaluate the tube formation and proliferation of HRECs within the stimulation of human recombinant BV8 protein.At the same time,the possible mechanism of BV8 promoting RNV formation was explored.Results: The expression level of BV8 was significantly increased in OIR mice and Rho/VEGF transgenic mice.After intravitreal injection of anti-BV8 neutralizing antibody,retinal and sub-retinal neovascularization were inhibited,and the expression level of pro-angiogenic factors was significantly decreased.The expression of BV8 was increased in HRECs in hypoxia.Recombinant BV8 protein promoted vascular endothelial cell tube-formation and cell proliferation.BV8 affected RNV formation by regulating MAPK and AKT signaling pathway.Conclusions: BV8 promotes RNV by regulating endothelial cells and can be used as a new therapeutic target for the treatment of RNV diseases.