Department of Ophthalmology, University Medical Center, University of Kiel, Arnold-Heller-Str. 3, Haus 25, 24105 Kiel, Germany.
Alginor ASA, Haraldsgata 162, 5525 Haugesund, Norway.
Int J Mol Sci. 2023 Jan 23;24(3):2232. doi: 10.3390/ijms24032232.
Fucoidans are polysaccharides and constituents of cell walls of brown algae such as (LH). They exhibit promising effects regarding age-related macular degeneration (AMD). However, the safety of this compound needs to be assured. The focus of this study lies on influences of an LH fucoidan on the retinal pigment epithelium (RPE). The high-molecular weight LH fucoidan Fuc1 was applied to primary porcine RPE cells, and a tetrazolium (MTT) cell viability assay was conducted. Further tests included a scratch assay to measure wound healing, Western blotting to measure expression of retinal pigment epithelium-specific 65 kDa protein (RPE65), as well as immunofluorescence to measure uptake of opsonized fluorescence beads into RPE cells. Lipopolysaccharide was used to proinflammatorily activate the RPE, and interleukin 6 (IL-6) and interleukin 8 (IL-8) secretion was measured. RPE/choroid cultures were used to assess vascular endothelial growth factor (VEGF) secretion. Real-time polymerase chain reaction (real-time PCR) was performed to detect the gene expression of 91 different genes in a specific porcine RPE gene array. Fuc1 slightly reduced wound healing, but did not influence cell viability, phagocytosis or RPE65 expression. Fuc1 lowered IL-6, IL-8 and VEGF secretion. Furthermore, Fuc1 did not change tested RPE genes. In conclusion, Fuc1 does not impair RPE cellular functions and shows antiangiogenic and anti-inflammatory activities, which indicates its safety and strengthens its suitability concerning ocular diseases.
褐藻中的岩藻依聚糖是多糖和细胞壁的组成部分,如(LH)。它们在年龄相关性黄斑变性(AMD)方面显示出有希望的效果。然而,这种化合物的安全性需要得到保证。本研究的重点在于 LH 岩藻依聚糖对视网膜色素上皮(RPE)的影响。高分子量 LH 岩藻依聚糖 Fuc1 被应用于原代猪 RPE 细胞,并进行了噻唑蓝(MTT)细胞活力测定。进一步的测试包括划痕试验以测量伤口愈合,Western blot 测定视网膜色素上皮特异性 65 kDa 蛋白(RPE65)的表达,以及免疫荧光测定吞噬作用被调理的荧光珠进入 RPE 细胞。脂多糖被用来使 RPE 产生炎症激活,并且测量白细胞介素 6(IL-6)和白细胞介素 8(IL-8)的分泌。RPE/脉络膜培养物用于评估血管内皮生长因子(VEGF)的分泌。通过实时聚合酶链反应(real-time PCR)在特定的猪 RPE 基因阵列中检测 91 个不同基因的基因表达。Fuc1 轻微地降低了伤口愈合,但不影响细胞活力、吞噬作用或 RPE65 的表达。Fuc1 降低了 IL-6、IL-8 和 VEGF 的分泌。此外,Fuc1 没有改变测试的 RPE 基因。总之,Fuc1 不会损害 RPE 细胞功能,并表现出抗血管生成和抗炎活性,这表明其安全性,并增强了其在眼部疾病方面的适用性。