Department of Ophthalmology, University Medical Center, University of Kiel, Arnold-Heller-Str. 3, Haus 25, 24105 Kiel, Germany.
Pharmaceutical Institute, University of Kiel, Gutenbergstr. 76, 24118 Kiel, Germany.
Int J Mol Sci. 2023 Apr 27;24(9):7939. doi: 10.3390/ijms24097939.
Fucoidans from brown algae are described as anti-inflammatory, antioxidative, and antiangiogenic. We tested two fucoidans (SL-FRO and SL-NOR) regarding their potential biological effects against age-related macular degeneration (AMD). Primary porcine retinal pigment epithelium (RPE), human RPE cell line ARPE-19, and human uveal melanoma cell line OMM-1 were used. Cell survival was assessed in tetrazolium assay (MTT). Oxidative stress assays were induced with erastin or HO. Supernatants were harvested to assess secreted vascular endothelial growth factor A (VEGF-A) in ELISA. Barrier function was assessed by measurement of trans-epithelial electrical resistance (TEER). Protectin (CD59) and retinal pigment epithelium-specific 65 kDa protein (RPE65) were evaluated in western blot. Polymorphonuclear elastase and complement inhibition assays were performed. Phagocytosis of photoreceptor outer segments was tested in a fluorescence assay. Secretion and expression of proinflammatory cytokines were assessed with ELISA and real-time PCR. Fucoidans were chemically analyzed. Neither toxic nor antioxidative effects were detected in ARPE-19 or OMM-1. Interleukin 8 gene expression was slightly reduced by SL-NOR but induced by SL-FRO in RPE. VEGF secretion was reduced in ARPE-19 by SL-FRO and in RPE by both fucoidans. Polyinosinic:polycytidylic acid induced interleukin 6 and interleukin 8 secretion was reduced by both fucoidans in RPE. CD59 expression was positively influenced by fucoidans, and they exhibited a complement and elastase inhibitory effect in cell-free assay. RPE65 expression was reduced by SL-NOR in RPE. Barrier function of RPE was transiently reduced. Phagocytosis ability was slightly reduced by both fucoidans in primary RPE but not in ARPE-19. Fucoidans from , especially SL-FRO, are promising agents against AMD, as they reduce angiogenic cytokines and show anti-inflammatory and complement inhibiting properties; however, potential effects on gene expression and RPE functions need to be considered for further research.
褐藻来源的岩藻聚糖硫酸酯被描述为具有抗炎、抗氧化和抗血管生成作用。我们测试了两种岩藻聚糖硫酸酯（SL-FRO 和 SL-NOR），以评估其对年龄相关性黄斑变性（AMD）的潜在生物学作用。使用原代猪视网膜色素上皮（RPE）、人 RPE 细胞系 ARPE-19 和人葡萄膜黑色素瘤细胞系 OMM-1。通过四唑盐（MTT）测定法评估细胞存活率。用 erastin 或 HO 诱导氧化应激试验。收集上清液，通过 ELISA 评估分泌的血管内皮生长因子 A（VEGF-A）。通过测量跨上皮电阻（TEER）评估屏障功能。通过 Western blot 评估保护蛋白（CD59）和视网膜色素上皮特异性 65kDa 蛋白（RPE65）。进行多形核弹性蛋白酶和补体抑制试验。在荧光测定中测试光感受器外节的吞噬作用。通过 ELISA 和实时 PCR 评估促炎细胞因子的分泌和表达。对岩藻聚糖硫酸酯进行了化学分析。在 ARPE-19 或 OMM-1 中均未检测到毒性或抗氧化作用。SL-NOR 略微降低了 RPE 中的白细胞介素 8 基因表达，但 SL-FRO 诱导了其表达。SL-FRO 降低了 ARPE-19 中的 VEGF 分泌，两种岩藻聚糖硫酸酯均降低了 RPE 中的 VEGF 分泌。多聚肌苷酸：多聚胞苷酸诱导的白细胞介素 6 和白细胞介素 8 分泌均被两种岩藻聚糖硫酸酯在 RPE 中降低。CD59 表达受岩藻聚糖硫酸酯的正向影响，并且它们在无细胞测定中具有补体和弹性蛋白酶抑制作用。SL-NOR 降低了 RPE 中的 RPE65 表达。RPE 的屏障功能短暂降低。两种岩藻聚糖硫酸酯均略微降低了原代 RPE 中的吞噬能力，但对 ARPE-19 没有影响。岩藻聚糖硫酸酯，尤其是 SL-FRO，是对抗 AMD 的有前途的药物，因为它们可减少血管生成细胞因子并具有抗炎和补体抑制作用；然而，需要进一步研究其对基因表达和 RPE 功能的潜在影响。
