Wu L, Tan X, Liang L, Yu H, Wang C, Zhang D, Kijlstra A, Yang P
The First Affiliated Hospital of Chongqing Medical University, Chongqing Key Lab of Ophthalmology, Chongqing Eye Institute, Chongqing. China.
Eye Research Institute Maastricht, Department of Ophthalmology, University Hospital Maastricht, Maastricht. Netherlands.
Curr Mol Med. 2017;17(2):140-148. doi: 10.2174/1566524017666170331162616.
This study aimed to investigate the mechanisms whereby Amyloidbeta (Aβ) induces the production of angiogenic factors by a human retinal pigment epithelial cell line (ARPE-19) cells.
ARPE-19 cells obtained from the American Type Culture Collection (ATCC) were utilized in this study. The expression level of vascular endothelial growth factor (VEGF), Interleukin-8 (IL-8), monocyte chemoattractant protein-1 (MCP-1) and complement activation fragments C3a and C5a were measured by Real-time quantitative PCR (RT-PCR) and Enzyme-linked immunosorbent assay (ELISA). The production of mitochondria-associated reactive oxygen species (ROS) was measured by flow cytometry.
The expression of VEGF, IL-8, MCP-1, C3a and C5a was significantly increased in Aβ-treated ARPE-19 cells. Mitochondria-associated ROS production was also significantly increased when exposed to Aβ. Inhibition of mitochondrial ROS with Diphenyleneiodonium chloride (DPI) markedly decreased the Aβ induced production of VEGF, IL-8, MCP-1, C3a and C5a by ARPE-19 cells. Anti-C3a or anti-C5a neutralizing antibodies did not have a detectable influence on the secretion of VEGF, IL-8 and MCP-1 by ARPE-19 cells upon stimulation with Aβ.
Our results support the hypothesis that Aβ is involved in the pathogenesis of choroidal neovascularization (CNV) formation by promoting the production of the angiogenic cytokines VEGF, IL-8 and MCP-1 by RPE cells. Mitochondrial ROS was shown to play a role in the regulation of Aβ induced expression of these cytokines.
本研究旨在探究淀粉样β蛋白（Aβ）诱导人视网膜色素上皮细胞系（ARPE - 19）细胞产生血管生成因子的机制。
本研究使用了从美国典型培养物保藏中心（ATCC）获得的ARPE - 19细胞。通过实时定量聚合酶链反应（RT - PCR）和酶联免疫吸附测定（ELISA）测量血管内皮生长因子（VEGF）、白细胞介素 - 8（IL - 8）、单核细胞趋化蛋白 - 1（MCP - 1）以及补体激活片段C3a和C5a的表达水平。通过流式细胞术测量线粒体相关活性氧（ROS）的产生。
在Aβ处理的ARPE - 19细胞中，VEGF、IL - 8、MCP - 1、C3a和C5a的表达显著增加。暴露于Aβ时，线粒体相关ROS的产生也显著增加。用二苯基碘鎓氯化物（DPI）抑制线粒体ROS可显著降低Aβ诱导的ARPE - 19细胞产生VEGF、IL - 8、MCP - 1、C3a和C5a。抗C3a或抗C5a中和抗体对Aβ刺激后ARPE - 19细胞分泌VEGF、IL - 8和MCP - 1没有可检测到的影响。
我们的结果支持以下假设，即Aβ通过促进视网膜色素上皮（RPE）细胞产生血管生成细胞因子VEGF、IL - 8和MCP - 1参与脉络膜新生血管（CNV）形成的发病机制。线粒体ROS在调节Aβ诱导的这些细胞因子表达中发挥作用。
