Department of Anatomy, University of Pécs, Medical School, MTA-PTE PACAP Research Group, Pécs, Hungary.
Department of Medical Chemistry, University of Szeged, Szeged, Hungary.
Ann N Y Acad Sci. 2019 Nov;1455(1):160-172. doi: 10.1111/nyas.14189. Epub 2019 Jul 17.
The purpose of this study was to determine whether pituitary adenylate cyclase activating polypeptide (PACAP) could influence the neovascularization processes in hyperosmotic and oxidative stress in retinal pigment epithelial cells. Hyperosmotic conditions and oxidative stress were induced by 200 mM sucrose and 250 µM hydrogen peroxide (H O ), respectively. Morphology and elasticity of adult retinal pigment epithelial (ARPE-19) cells were measured by atomic force microscopy, while the investigation of junctional molecules, such as occludin and ZO-1, was carried out using immunofluorescence. For cell viability measurement, the MTT test was used. The effect of PACAP on the key angiogenic factors, such as vascular endothelial growth factor, angiogenin, and endothelin-1, was measured by an angiogenesis array and flow cytometry. Hyperosmotic stress-induced reorganization of the cytoskeleton and impairment of the junctions decreased cell viability and upregulated several angiogenic factors. In oxidative stress, we found that opening of the junctions decreased viability and upregulated the expression of angiogenic factors. PACAP was shown to be protective in both conditions. Retinal pigment epithelium cells play an important role in several diseases, such as diabetic retinopathy and macular edema. Therefore, protecting retinal pigment epithelial (RPE) cells with PACAP could be a novel and potential treatment in these diseases.
本研究旨在探讨垂体腺苷酸环化酶激活肽（PACAP）是否能影响高渗和氧化应激条件下视网膜色素上皮细胞的血管新生过程。通过 200mM 蔗糖和 250μM 过氧化氢（H₂O₂）分别诱导高渗和氧化应激。原子力显微镜测量成年视网膜色素上皮（ARPE-19）细胞的形态和弹性，免疫荧光法检测连接分子如闭合蛋白和 ZO-1。采用 MTT 试验检测细胞活力。通过血管生成阵列和流式细胞术检测 PACAP 对血管内皮生长因子、血管生成素和内皮素-1 等关键血管生成因子的影响。高渗应激诱导的细胞骨架重排和连接破坏降低了细胞活力并上调了几种血管生成因子。在氧化应激中，我们发现连接的开放降低了细胞活力并上调了血管生成因子的表达。PACAP 在这两种情况下均具有保护作用。视网膜色素上皮细胞在多种疾病中起重要作用，如糖尿病视网膜病变和黄斑水肿。因此，用 PACAP 保护视网膜色素上皮（RPE）细胞可能是这些疾病的一种新的潜在治疗方法。
