Mechanism Of PiRNA-1245/PIWIL2 Regulating JAK2/STAT3/VEGF Signaling Pathway In Retinal Neovascularization

Objective: Retinal neovascularization is a common pathological change in many retinal diseases.Although the current use of anti VEGF drugs can effectively inhibit the formation of neovascularization,the resistance and damage to the retina caused by repeated use of anti VEGF drugs are irreversible.At present,researchers have studied the pathogenesis of retinal neovascularization from the perspectives of genomics,genetics and epigenetics.The research on the pathogenesis of retinal neovascularization has become the key to diagnosis and targeted therapy.Noncoding RNAs are the transcripts of most genes in human genome,which are generally divided into short noncoding RNAs and long noncoding RNAs.Short non coding RNAs mainly include micro RNAs,short interfering RNAs and piwi interacting RNAs.In the first part of this study,we found that piRNA was differentially expressed in the retina of mice with oxygen induced retinopathy,and piRNA-1245 was highly expressed in the retina of mice with oxygen induced retinopathy.In the second part,we first confirmed that piRNA-1245 and PIWIL2 protein were highly expressed in hypoxia treated human retinal endothelial cells,and knockdown of piRNA-1245 could reduce the expression of VEGF.At the same time,functional experiments confirmed that piRNA-1245 affected the migration,apoptosis and tube forming ability of hypoxic HREC cells.The third part is to verify whether piRNA-1245 can regulate retinal neovascularization through animal experiments.Methods: The expression of piRNA-1245 was detected by Real-time PCR,and the expression of PIWIL2 and VEGF was detected by Western blot,Pi R-1245 knockdown cell line was constructed by cell transfection method,and the changes of cell proliferation,migration,apoptosis and tubulation function were observed.Lentivirus was injected into the vitreous cavity of mice with oxygen induced retinopathy to observe the retinopathy and the expression of inflammatory factors.Results: In OIR group,79 piRNAs were differentially expressed,of which 43 were up-regulated and 36 were down regulated.Bioinformatics analysis showed that some differentially expressed genes were involved in the regulation of neovascularization,and some genes were involved in the disease through multiple pathways.PIWIL2 and piR-1245 were high expressed in the retina of OIR mice and hypoxic HREC cells.Knockdown of piR-1245 could reduce the expression of VEGF.Pi R-1245 knockdown can reduce the proliferation of hypoxic HREC,piR-1245 knockdown can reduce the migration and tube forming ability of hypoxic HREC,piR-1245 knockdown can inhibit the apoptosis of hypoxic HREC.Pi R-1245,PIWIL2 and VEGF were highly expressed in the retina of OIR mice.Intravitreal injection of piR-1245 lentivirus can reduce the inflammatory response of retinal tissue,and intravitreal injection of piR-1245 lentivirus can inhibit retinal neovascularization.Pi R-1245 activates JAK2 / STAT3 pathway to regulate the expression of VEGF,while piR-1245 activates JAK2 / STAT3 pathway to regulate the expression of VEGF and the formation of retinal neovascularization.Conclusion: piR-1245 is highly expressed in the retina of OIR mice and hypoxia treated HREC cells.Pi R-1245 knockdown can reduce the apoptosis rate of cells and enhance the migration,proliferation and tube forming ability of cells.Intravitreal injection of piR-1245 can inhibit retinal neovascularization,which is expected to become an important method for the treatment of retinal neovascularization.