ObjectiveTo investigate the effects of thrombospondin-1 active fragment (TSP-1) synthetical peptide VR-10 on proliferation and migration of rhesus choroidal-retinal endothelial (RF/6A) cell and the expressions of apoptosis relative genes in RF/6A cell. MethodsThe survival rate of RF/6A cell were detected by methyl thiazolyl tetrazolium, and migration ability was measured by transwell chamber after exposure to 1.0 μg/ml TSP-1 and synthetic peptide VR-10 (0.1, 1.0, 10.0 μg/ml) for different times (6, 12, 24, 48 hours). Caspase-3 and factor associated suicide (FAS) protein levels were measured by Western blot. The mRNA level of bcl-2 and FAS ligand (FASL) were measured by reverse transcription-polymerase chain reaction (RT-PCR). ResultsThe survival rate of RF/6A cells was determined by the treatment time and concentration of TSP-1(1.0 μg/ml) and the synthetic peptide VR-10 (0.1, 1.0, 10.0 μg/ml). The lowest survival ratio of RF/6A was 78% (P < 0.001) when cells were treated by 10 μg/ml synthetic peptide VR-10 after 48 hours. TSP-1 and synthetic peptide VR-10 could inhibit migration of RF/6A cells in transwell chamber (P < 0.001). 10.0 μg/ml synthetic peptide VR-10 had the strongest effect, 1.0 μg/ml TSP-1 was the next. Migration inhibition rate was increase with the increase of the concentration of VR-10 (P < 0.001). There was no significant differences between 0.1 μg/ml and 1.0 μg/ml VR-10 (P=0.114). Western bolt showed that RF/6A cell in control group mainly expressed the 32×103 procaspase-3 forms. To 10.0 μg/ml VR-10 treated group, it showed decreased expression of procaspase-3 (32×103) and concomitant increased expression of its shorter proapoptotic forms (20×103). Compared with control group, expression of FAS peptides were significantly increased in 10.0 μg/ml VR-10 treated group. Compared with control group, expression of FasL mRNA was significantly increased in 10.0 μg/ml VR-10 treated group(t=39.365, P=0.001), but the expression of bcl-2 mRNA was decreased(t=-67.419, P=0.000). ConclusionTSP-1 and synthetic peptide VR-10 had the ability to inhibit proliferation and migration of endothelial cell, and also induce apoptosis by increasing FAS/FASL expression and repressing bcl-2 expression.
Objective To study and compare the clinical efficacy between intravitreal conbercept injection and (or) macular grid pattern photocoagulation in treating macular edema secondary to non-ischemic branch retinal vein occlusion (BRVO). Methods Ninety eyes of 90 patients diagnosed as macular edema secondary to non-ischemic BRVO were enrolled in this study. Forty-eight patients (48 eyes) were male and 42 patients (42 eyes) were female. The average age was (51.25±12.24) years and the course was 5–17 days. All patients were given best corrected visual acuity (BCVA), intraocular pressure, slit lamp with preset lens, fluorescence fundus angiography (FFA) and optic coherent tomography (OCT) examination. The patients were divided into conbercept and laser group (group Ⅰ), laser group (group Ⅱ) and conbercept group (group Ⅲ), with 30 eyes in each group. The BCVA and central macular thickness (CMT) in the three groups at baseline were statistically no difference (F=0.072, 0.286;P=0.930, 0.752). Patients in group Ⅰ received intravitreal injection of 0.05 ml of 10.00 mg/ml conbercept solution (conbercept 0.5 mg), and macular grid pattern photocoagulation 3 days later. Group Ⅱ patients were given macular grid pattern photocoagulation. Times of injection between group Ⅰ and Ⅲ, laser energy between group Ⅰ and Ⅱ, changes of BCVA and CMT among 3 groups at 1 week, 1 month, 3 months and 6 months after treatment were compared. Results Patients in group Ⅰ and Ⅲ had received conbercept injections (1.20±0.41) and (2.23±1.04) times respectively, and 6 eyes (group Ⅰ) and 22 eyes (group Ⅲ) received 2-4 times re-injections. The difference of injection times between two groups was significant (P＜0.001). Patients in group Ⅱ had received photocoagulation (1.43±0.63) times, 9 eyes had received twice photocoagulation and 2 eyes had received 3 times of photocoagulation. The average laser energy was (96.05±2.34) μV in group Ⅰ and (117.41±6.85) μV in group Ⅱ, the difference was statistical significant (P=0.003). BCVA improved in all three groups at last follow-up. However, the final visual acuity in group Ⅰ and group Ⅲ were better than in group Ⅱ (t=4.607, –4.603;P＜0.001) and there is no statistical significant difference between group Ⅲ and group Ⅰ (t=–0.802,P=0.429). The mean CMT reduced in all three groups after treating for 1 week and 1 month, comparing that before treatment (t=–11.855, –10.620, –10.254;P＜0.001). There was no statistical difference of CMT between group Ⅰand Ⅲ at each follow up (t=0.404, 1.723, –1.819, –1.755;P=0.689, 0.096, 0.079, 0.900). CMT reduction in group Ⅰ was more than that in group Ⅱ at 1 week and 1 month after treatments (t=–4.621, –3.230;P＜0.001, 0.003). The CMT in group Ⅲ at 3 month after treatment had increased slightly comparing that at 1 month, but the difference was not statistically significant (t=1.995,P=0.056). All patients had no treatment-related complications, such as endophthalmitis, rubeosis iridis and retinal detachment. Conclusions Intravitreal conbercept injection combined with macular grid pattern photocoagulation is better than macular grid pattern photocoagulation alone in treating macular edema secondary to non-ischemic BRVO. Combined therapy also reduced injection times comparing to treatment using conbercept injection without laser photocoagulation.