Objective To investigate the effects of heparanase and vascular endothelial growth factor (VEGF) and their correlation in CoCl2 induced human retinal microvascular endothelial cells (HRECs) in an hypoxia model. Methods Human eyes were selected to establish CoCl2induced HRECs hypoxia model in this study. Four experimental groups were studied: normal control group, hypoxia group (CoCl2 100 μmol/L, 48 hours),PI-88 group (specific competitive inhibitor of heparanase: phosphomannopentaose sulfate, PI-88,5 μg/ml, combined with CoCl2 100 μmol/L, 48 hours) and PBS control group. Heparanase, VEGF and Pol Ⅱ expression in HRECs of normal and hypoxia group were analyzed with immunofluorescence. Western blot was used to evaluate the expression of heparanase and VEGF in HRECs of normal, hypoxia, PI88 and PBS control groups. ResultsImmunofluorescence studies showed that the expression of heparanase and VEGF in cytoplasm was intense in hypoxia HRECs, but faint in normal group. Heparanase was also observed in the nucleus of hypoxia HRECs. Western blot results showed that the expression of Hpa and VEGF protein was increased significantly in hypoxia group compared with normal group (Hpa：F=-4。005, P＜0.05；VEGF：F=-4.063, P＜0.05), and VEGF was decreased in HRECs treated with PI-88(F=5。963, P＜0.05). ConclusionsHeparanase is upregulated that resulted in increase of VEGF expression, therefore enhanced angiogenesis in CoCl2 induced hypoxia HRECs.
Objective To observe the efficacy of monotherapy and combination treatments of photodynamic therapy (PDT) and intravitreal anti-vascular endothelial growth factor (VEGF) drugs for advanced exudative age-related macular degeneration (EAMD). Methods Thirty-two eyes from 28 patients with EAMD were enrolled in this study. Fifteen eyes were treated with PDT or intravitreal anti-VEGF drugs (monotherapy group), while 17 eyes were treated with combination treatments of PDT and intravitreal anti-VEGF drugs (combination group). The follow-up was ranged from 5 to 28 months with a mean of 21 months. Vision and intraocular pressure were measured and optical coherence tomography (OCT) was performed every month after treatment. Fundus fluorescein angiography (FFA) and indocyanine green angiography (ICGA) were performed every 3 months during follow-up period. The changes of visual acuity, leakage in macula lutea and central retinal thickness (CRT) before and after treatment, and adverse events during follow-up period were observed. The mean treatment times of monotherapy and combination group were compared and analyzed. Results Among 32 eyes, visual acuity improved in 20 eyes (62.5%), stabilized in 9 eyes (28.1%) and decreased in 3 eyes (9.4%). The leakage in macula lutea disappeared in 17 eyes (53.1%), decreased in 12 eyes (37.5%) and increased in 3 eyes (9.4%). CRT was reduced in 23 eyes (71.8%), increased in 9 eyes (28.2%). The mean treatment times of the combination group (2.3plusmn;0.2) was significantly less than that of monotherapy group (3.4plusmn;0.5)(P＜0.05). No adverse events such as secondary retinal hemorrhage, vitreous hemorrhage or endoophthalmitis were found during the follow-up duration. Conclusions Both monotherapy and combination treatments of PDT and intravitreal anti-VEGF drugs for EAMD may improve visual acuity, exudation and macular edema. Combination of PDT and anti-VEGF therapy decreases the frequency of retreatment.