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目的 比较不同药理批次重组诱饵型受体创新药物RC28-E的体外药效学,验证工艺变更前后生物治疗药物的效果是否存在差异.方法 将RF/6A细胞分为正常对照组、血管内皮生长因子(VEGF)+成纤维细胞生长因子(FGF)组和不同质量浓度RC28-E1组,采用细胞计数试剂盒8(CCK-8)法筛选出RC28-E1最适作用浓度.将细胞分为正常对照组、VEGF+FGF组、RC28-E1处理组、RC28-E2处理组、康柏西普处理组和FGF trap处理组,分别用无血清正常培养液、含VEGF+FGF无血清培养液、含VEGF+FGF+RC28-E1无血清培养液、含VEGF+FGF+RC28-E2的无血清培养液、含VEGF+FGF+康柏西普无血清培养液以及含VEGF+FGF+FGF trap无血清培养液进行培养.采用CCK-8法检测各组细胞增生率;采用Transwell试验检测各组细胞迁移能力;采用Matrigel实验检测各组细胞管腔形成能力. 结果 正常对照组、VEGF+FGF组、0.025 mg/mlRC28-E1组、0.080 mg/ml RC28-E1组和0.240 mg/ml RC28-E1组细胞增生率总体比较差异有统计学意义(F=6.601,P=0.012);其中0.080 mg/ml RC28-E1组细胞增生率明显低于VEGF+FGF组,差异有统计学意义(P＜0.05),以0.080 mg/ml为RC28-E最适工作浓度.正常对照组、VEGF+FGF组、RC28-E1处理组、RC28-E2处理组、康柏西普处理组、FGF trap处理组细胞增生率总体比较,差异有统计学意义(F=3.210,P=0.019);其中RC28-E1处理组、RC28-E2处理组和FGF trap处理组的细胞增生率均明显低于VEGF+FGF组,差异均有统计学意义(均P＜0.05).各组移行细胞数目总体比较差异有统计学意义(F=24.640,P=0.000);其中RC28-E1处理组、RC28-E2处理组、康柏西普处理组及FGF trap处理组移行细胞数均明显低于VEGF+FGF组,差异均有统计学意义(均P=0.000);RC28-E2处理组移行细胞数少于RC28-E1处理组,差异有统计学意义(P=0.002).各组细胞管腔形成数总体比较,差异有统计学意义(F=9.273,P=0.000),其中RC28-E1处理组、RC28-E2处理组、康柏西普处理组及FGF trap处理组内皮细胞管腔形成数均明显低于VEGF+FGF组,差异均有统计学意义(P=0.003、0.001、0.009、0.018);RC28-E2处理组细胞管腔形成数与正常对照组、RC28-E1处理组和康柏西普处理组相比,差异均无统计学意义(均P＞0.05);FGF trap处理组细胞管腔形成数明显高于RC28-E1处理组、RC28-E2处理组、康柏西普组及正常对照组,差异均有统计学意义(P=0.014、0.000、0.008、0.014). 结论 在体外VEGF+FGF刺激下,RC28-E对视网膜血管内皮细胞的增生抑制效果优于康柏西普,对管腔形成能力的抑制作用优于FGF trap.不同批次的重组诱饵型受体创新药物在视网膜血管内皮细胞中的效果无显著差异.“,”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-E 1 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.