Vanderbilt Eye Institute, Vanderbilt University Medical Center, Nashville, TN, USA.
Vanderbilt Eye Institute, Vanderbilt University Medical Center, Nashville, TN, USA.
Exp Eye Res. 2020 Apr;193:107974. doi: 10.1016/j.exer.2020.107974. Epub 2020 Feb 14.
Previous metabolomics studies from our lab found altered plasma levels of bile acids in patients with age-related macular degeneration (AMD) compared to controls. In this study, we investigated the ability of the bile acid taurocholic acid (TCA) to inhibit features of AMD modeled in vitro. Paraquat was used to induce oxidative stress in HRPEpiC primary retinal pigment epithelial (RPE) cells. Cells were treated with 300 μM paraquat alone or with TCA (10, 50, 100, 200, or 500 μM). RPE tight junction integrity was assessed via ZO-1 immunofluorescence and transepithelial electrical resistance (TEER) measurements. RF/6A macaque choroidal endothelial cells were treated with 100 ng/mL vascular endothelial growth factor (VEGF) to induce angiogenesis. The effect of TCA on VEGF-induced angiogenesis was evaluated with cell proliferation, cell migration, and tube formation assays. Addition of TCA at 100 (P = 8.6 × 10), 200 (P = 0.0035), and 500 (P = 2.1 × 10) μM resulted in significant preservation of TEER in paraquat treated cells. In RF/6A cells, TCA did not significantly affect VEGF-induced cell proliferation. VEGF-induced migration of RF/6A cells was significantly inhibited at TCA concentrations of 100 (P = 0.010), 200 (P = 0.023) and 500 (P = 0.0049) μM. VEGF-induced tube formation was significantly inhibited when treated with 200 (P = 0.014) and 500 (P = 7.1 × 10) μM TCA. In vitro, TCA promoted RPE cell integrity and diminished VEGF-induced choroidal endothelial cell migration and tube formation. This suggests that TCA may have protective effects against both degenerative and neovascular AMD.
先前我们实验室的代谢组学研究发现，与对照相比，年龄相关性黄斑变性（AMD）患者的血浆胆汁酸水平发生了改变。在这项研究中，我们研究了胆汁酸牛磺胆酸（TCA）抑制体外模拟 AMD 特征的能力。百草枯用于诱导 HRPEpiC 原代视网膜色素上皮（RPE）细胞的氧化应激。用 300μM 百草枯单独或用 TCA（10、50、100、200 或 500μM）处理细胞。通过 ZO-1 免疫荧光和跨上皮电阻（TEER）测量评估 RPE 紧密连接完整性。用 100ng/mL 血管内皮生长因子（VEGF）处理 RF/6A 猕猴脉络膜内皮细胞以诱导血管生成。用细胞增殖、细胞迁移和管形成测定评估 TCA 对 VEGF 诱导的血管生成的影响。在百草枯处理的细胞中，TCA 浓度为 100（P=8.6×10）、200（P=0.0035）和 500（P=2.1×10）μM 时，TEER 显著得到保留。在 RF/6A 细胞中，TCA 对 VEGF 诱导的细胞增殖没有显著影响。TCA 浓度为 100（P=0.010）、200（P=0.023）和 500（P=0.0049）μM 时，VEGF 诱导的 RF/6A 细胞迁移显著受到抑制。当用 200（P=0.014）和 500（P=7.1×10）μM TCA 处理时，VEGF 诱导的管形成显著受到抑制。在体外，TCA 促进 RPE 细胞完整性，并减少 VEGF 诱导的脉络膜内皮细胞迁移和管形成。这表明 TCA 可能对退行性和新生血管性 AMD 均具有保护作用。
