ABSTRACT
Objective To compare the protective effects of pharmacological batch RC28-E1 and pilot batch RC28-E2 on retinal vascular endothelial cells (RF/6A) under the stimulation of vascular endothelial growth factor (VEGF) and fibroblast growth factor (FGF).Methods RF/6A cells were divided into normal control group,VEGF + FGF group and RC28-E1 groups with different concentrations.The optimal concentration of RC28-E1 was determined by cell counting kit-8 (CCK-8) method.Cells were divided into normal control group,VEGF+FGF group,RC28-E1 group,RC28-E2 group,conbercept group and FGF trap group,and cultured with serum-free culture medium,serum-free culture medium containing VEGF+FGF,serum-free culture medium containing VEGF+FGF+RC28-E1,serum-free culture medium containing VEGF+FGF+RC28-E2,and serum-free culture medium containing VEGF+FGF+ conbercept,serum-free medium containing VEGF+FGF+FGF trap,respectively.Cell proliferation rate was measured by CCK-8 method,cell migration ability was detected by Transwell test,and tube formation ability was detected by Matrigel assay.Results The cell proliferation rate of 0.080 mg/ml RC28-E1 group was significantly lower than that of VEGF+FGF group (P＜0.05).The cell proliferation rate of RC28-E1 group,RC28-E2 group and FGF trap group were significantly lower than that of VEGF+FGF group (P＜0.05).The number of migrated cells in RC28-E1 group,RC28-E2 group,conbercept group and FGF trap group were significantly lower than that in VEGF+FGF group (P=0.000).The numbers of meshes formed by retinal vascular endothelial cells in RC28-E1 group,RC28-E2 group,conbercept group and FGF trap group were significantly lower than that in VEGF+FGF group (P =0.003,0.001,0.009,0.018).The number of tube formation in FGF trap group was significantly higher than those in RC28-E1 group,RC28-E2 group,conbercept group and normal control group (P =0.014,0.000,0.008,0.014).Conclusions Under the stimulation of VEGF + FGF,the inhibitory effect of RC28-E on the proliferation of retinal vascular endothelial cells is greater than that of conbercept,and its inhibitory effect on the tube formation is superior to that of FGF trap.There is no significant difference in the effects of different batches of recombinant decoy receptor innovative drugs on retinal vascular endothelial cells.
ABSTRACT
Objective To compare the protective effects of pharmacological batch RC28.E1 and pilot batch RC28.E2 on retinal vascular endothelial cells ( RF/6A) under the stimulation of vascular endothelial growth factor (VEGF) and fibroblast growth factor (FGF). Methods RF/6A cells were divided into normal control group, VEGF + FGF group and RC28.E1 groups with different concentrations. The optimal concentration of RC28.E1 was determined by cell counting kit.8 (CCK.8) method. Cells were divided into normal control group,VEGF+FGF group, RC28.E1 group,RC28.E2 group,conbercept group and FGF trap group,and cultured with serum.free culture medium, serum.free culture medium containing VEGF+FGF,serum.free culture medium containing VEGF+FGF+RC28.E1, serum.free culture medium containing VEGF+FGF+RC28.E2,and serum.free culture medium containing VEGF+FGF+conbercept,serum.free medium containing VEGF+FGF+FGF trap,respectively. Cell proliferation rate was measured by CCK.8 method, cell migration ability was detected by Transwell test, and tube formation ability was detected by Matrigel assay. Results The cell proliferation rate of 0. 080 mg/ml RC28.E1 group was significantly lower than that of VEGF+FGF group (P<0. 05). The cell proliferation rate of RC28.E1 group,RC28.E2 group and FGF trap group were significantly lower than that of VEGF+FGF group (P<0. 05). The number of migrated cells in RC28.E1 group,RC28.E2 group,conbercept group and FGF trap group were significantly lower than that in VEGF+FGF group (P=0. 000). The numbers of meshes formed by retinal vascular endothelial cells in RC28.E1 group,RC28.E2 group, conbercept group and FGF trap group were significantly lower than that in VEGF+FGF group ( P=0. 003,0. 001, 0. 009,0. 018). The number of tube formation in FGF trap group was significantly higher than those in RC28.E1 group,RC28.E2 group, conbercept group and normal control group ( P = 0. 014, 0. 000, 0. 008, 0. 014 ). Conclusions Under the stimulation of VEGF+FGF,the inhibitory effect of RC28.E on the proliferation of retinal vascular endothelial cells is greater than that of conbercept,and its inhibitory effect on the tube formation is superior to that of FGF trap. There is no significant difference in the effects of different batches of recombinant decoy receptor innovative drugs on retinal vascular endothelial cells.