Kiel University, University Medical Center, Department of Ophthalmology, Arnold-Heller-Str. 3, Haus B2, 24105, Kiel, Germany.
Kiel University, University Medical Center, Department of Ophthalmology, Arnold-Heller-Str. 3, Haus B2, 24105, Kiel, Germany.
Exp Eye Res. 2023 Jun;231:109469. doi: 10.1016/j.exer.2023.109469. Epub 2023 Apr 8.
Age-related macular degeneration (AMD) is the leading cause of severe vision loss in industrialized nations. Important factors in pathogenesis are oxidative stress, inflammation, and, in the wet form of AMD, angiogenesis. Fucoidans, sulfated polysaccharides from brown algae, may have antioxidant, anti-inflammatory, and antiangiogenic effects. In this study, we established specific gene expression panels for inflammation, oxidative stress and angiogenesis in porcine retinal pigment epithelium (RPE), and investigated the effect of fucoidans on gene expression under different noxious agents.
Primary porcine RPE cells cultured for at least 14 days were used. Using viability assays with tetrazolium bromide and real-time polymerase chain reaction of marker genes, positive controls were established for appropriate concentrations and exposure times of selected noxious agents (lipopolysaccharide (LPS), HO, CoCl). Three different AMD relevant gene panels specific for porcine RPE for inflammation, oxidative stress, and angiogenesis were established, and the influence of fucoidans (mainly Fucus vesiculosus; FV) on gene expression was investigated.
The following was shown by gene expression analyses: (1) Inflammation panel: Expression of 18 genes was affected under LPS (three days). Among them, LPS increased genes for interleukin 1 receptor 2, interleukin 8, cyclooxygenase-2 and vascular cell adhesion protein 1 expression which were diminished when FV was present. (2) Oxidative stress panel: Under stimulation of HO (one day) and LPS (one day), expression of a total of 15 genes was affected. LPS induced increase in genes for superoxide dismutase-1, C-X-C motif chemokine 10, and CC chemokine ligand-5 expression was not detected when FV was present. (3) Angiogenesis panel: Under stimulation with CoCl (three days) expression of six genes was affected, with the increase of genes for angiopoietin 2, vascular endothelial growth factor receptor-1, and follistatin being diminished when FV was present.
Three specific gene expression panels for porcine RPE that map genes for three of the major pathological factors of AMD, inflammation, oxidative stress, and angiogenesis, were established. Further, we demonstrated that fucoidans can reduce stress related gene activation in all of these three major pathogenic pathways. This study is another indication that fucoidans can act on different pathomechanisms of AMD simultaneously, which provides further evidence for fucoidans as a possible drug for treatment and prevention of AMD.
年龄相关性黄斑变性（AMD）是工业化国家导致严重视力丧失的主要原因。发病机制中的重要因素是氧化应激、炎症，以及在湿性 AMD 中，血管生成。褐藻中的硫酸多糖褐藻胶可能具有抗氧化、抗炎和抗血管生成作用。在这项研究中，我们建立了猪视网膜色素上皮（RPE）中炎症、氧化应激和血管生成的特定基因表达谱，并研究了褐藻胶在不同有害物质作用下对基因表达的影响。
使用至少培养 14 天的原代猪 RPE 细胞。通过四唑溴盐的活力测定和选定有害物质（脂多糖（LPS）、HO、CoCl）的实时聚合酶链反应的标记基因，为适当的浓度和暴露时间建立了阳性对照。建立了三个针对猪 RPE 的特定 AMD 相关基因表达谱，用于炎症、氧化应激和血管生成，并研究了褐藻胶（主要是泡叶藻；FV）对基因表达的影响。
基因表达分析显示：（1）炎症谱：在 LPS 作用下（三天），18 个基因的表达受到影响。其中，LPS 增加了白细胞介素 1 受体 2、白细胞介素 8、环氧化酶-2 和血管细胞黏附蛋白 1 的表达，而当存在 FV 时，这些基因的表达减少。（2）氧化应激谱：在 HO（一天）和 LPS（一天）的刺激下，总共 15 个基因的表达受到影响。LPS 诱导的超氧化物歧化酶-1、C-X-C 基序趋化因子 10 和 CC 趋化因子配体-5 基因表达增加，当存在 FV 时，这些基因的表达未被检测到。（3）血管生成谱：在 CoCl（三天）的刺激下，六个基因的表达受到影响，当存在 FV 时，血管生成素 2、血管内皮生长因子受体-1 和卵泡抑素的基因表达增加减少。
建立了三个针对猪 RPE 的特定基因表达谱，这些谱图绘制了 AMD 的三个主要病理因素的基因，即炎症、氧化应激和血管生成。此外，我们证明褐藻胶可以减少这三种主要致病途径中与应激相关的基因激活。这项研究进一步表明，褐藻胶可以同时作用于 AMD 的不同发病机制，为褐藻胶作为 AMD 治疗和预防的潜在药物提供了进一步的证据。
