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Abstract：
Purpose/Aim of the study: Vascular endothelial growth factor (VEGF)-antagonists are given over long time periods in the clinic, but the long-term effects on retinal pigment epithelium (RPE) cells are not fully investigated. This study aims to investigate these effects with two clinical relevant VEGF antagonists, bevacizumab and aflibercept, on the function of primary RPE cells.Materials and Methods: All tests were conducted with primary porcine RPE. Cells were stimulated with bevacizumab or aflibercept (both 250 µg/ml) for 1 day, 7 days or 4 weeks. Cell viability was tested in MTT Assay. Secretion of TGF-ß was tested in ELISA, phagocytosis in a microscopic assay, migration in a scratch assay, and expression of RPE65 in Western blot. Barrier function was tested for bevacizumab in transwell-cultured cells by measuring transepithelial electrical resistance for up to 3 days.Results: Viability was reduced by both antagonists at all time points tested. TGF-ß secretion was not altered by any treatment. Phagocytosis was not significantly reduced by any treatment. Wound healing ability was not significantly altered by any treatment. The expression of RPE65 was reduced by bevacizumab but not aflibercept after 4 weeks. Transepithelial electrical resistance was not altered.Conclusions: Long-term treatment with anti VEGF may affect viability of RPE cells, and treatment with bevacizumab may have effects on RPE function in long-term treatment.
摘要：
研究的目的/目的：血管内皮生长因子（VEGF）拮抗剂在临床上长期使用，但对视网膜色素上皮（RPE）细胞的长期影响尚未完全研究。本研究旨在探讨两种临床相关的VEGF拮抗剂的作用,贝伐单抗和阿柏西普,对原代RPE细胞功能的影响。材料和方法:所有试验均用原代猪RPE进行。用贝伐单抗或阿柏西普（均为250μg/ml）刺激细胞1天，7天或4周。在MTT测定中测试细胞活力。在ELISA中检测TGF-β的分泌，在显微镜检测中的吞噬作用，在划痕试验中迁移，和RPE65在Westernblot中的表达。通过测量跨上皮电阻长达3天，在transwell培养的细胞中测试贝伐单抗的屏障功能。结果:在测试的所有时间点,两种拮抗剂均降低了活力。任何治疗均未改变TGF-β的分泌。吞噬作用未被任何治疗显著降低。任何治疗均未显著改变伤口愈合能力。4周后，贝伐单抗降低了RPE65的表达，但阿柏西普没有降低。跨上皮电阻没有改变。结论:抗VEGF长期治疗可能影响RPE细胞的活力,在长期治疗中，贝伐单抗治疗可能对RPE功能产生影响。
