| Objective:To investigate the expression of allograft inflammatory factor-1(AIF-1)in the pathogenesis of macular neovascularization(MNV)and its mechanism of action,and to provide new potential molecular targets for the study of MNV pathogenesis and its treatment.Methods:1.A mouse MNV model was established by fundus laser photocoagulation induction,and choroidal retinal tissues were taken on days 1,3,7,14 and 28 after successful laser induction,and the expression trends of AIF-1 and vascular endothelial growth factor(VEGF)were detected by Western Blot.and compared with normal control group.2.Immunohistochemical staining was performed to detect the expression of AIF-1 at day 7 after laser induction and compared with normal controls.3.On day 7 after laser induction,mice were grouped according to treatment:(1)normal control group;(2)laser day 7 injury control group;(3)intravitreal injection(LVI)ranibizumab(RBZ)group;(4)IVI AIF-1 small interfering RNA(siRNA)group;(5)IVI Scramble siRNA group;and(6)IVI phosphate buffered saline(PBS)group.The retinal choroidal tissues were removed from the eyes of each group under the microscope,and the effects of different treatments on the expression of AIF-1 and VEGF protein were examined by Western Blot.4.Fundus fluorescein angiography(FFA)and indocyanine green angiography(ICGA)were used to assess the effect of each group on the area of MNV leakage.Hematoxylin-eosin staining(HE)of frozen sections of mouse eyes was used to detect structural changes and integrity of the retina in the area of MNV lesions in different treatment groups.5.Choroidal pavement fluorescence staining was performed to assess the aggregation of choroidal endothelial cells(CECs)within the MNV lesion area in different treatment groups.6.After silencing of AIF-1 gene by IVI AIF-1 siRNA,the expression of P44/42 MAPK was detected by Western Blot and compared with the normal group and the laser-damaged control group.Results:1.In the mouse MNV model induced by laser injury,Western Blot results showed that the protein expression of AIF-1 and VEGF gradually increased with time after laser photocoagulation compared with normal controls(P<0.05),and the expression peaked at days 7-14 and gradually decreased afterwards,with the same expression trend for both.2.Immunohistochemistry results showed that the expression of AIF-1 was increased in the area of MNV lesions on day 7 after laser photocoagulation compared with normal controls.3.After successful laser photocoagulation modeling of mice,AIF-1 siRNA and RBZ were intravitreal injection immediately,and the results of Western Blot testing on day 7 showed that the protein expression of AIF-1 and VEGF were significantly reduced in the AIF-1 siRNA and RBZ groups compared with the injury control group(both P<0.05).4.The FFA and ICGA results showed that the leakage area of MNV was significantly reduced in the AIF-1 siRNA group and RBZ group;HE staining results also showed that both IVI AIF-1 siRNA and RBZ significantly reduced the area of MNV lesions.5.Fluorescence staining of choroidal pavement in mice showed a significant reduction in CECs aggregation in the area of MNV lesions in the AIF-1 siRNA group as well as in the RBZ group compared with the control in the 7-day laser injury group.6.IVI AIF-1 siRNA silencing of the AIF-1 gene followed by Western Blot assay showed that the expression of P44/42 MAPK was decreased compared to the damaged control group(P<0.05).Conclusion:AIF-1 plays an important role in the pathological progression of MNV.Inhibition of CECs via the p44/42 MAPK signaling pathway after intravitreal injection of AIF-1 siRNA to silence the AIF-1 gene significantly alleviated the pathological progression of MNV.As a new molecular target with therapeutic potential for MNV,the clinical application value of AIF-1 needs to be investigated more thoroughly. |
PDF Full Text Request