ABSTRACT

Background

Circular_0122396 (circ_0122396) has been reported to be downregulated in age-related cataract (ARC); however, the underlying mechanism remains unknown. The study aimed to reveal the role of circ_0122396 in ARC progression and underneath mechanism.

Methods

Hydrogen peroxide (H₂O₂) was employed to induce lens epithelial cells (SRA01/04) injury. The RNA expression of circ_0122396, microRNA-15a-5p (miR-15a-5p) and fibroblast growth factor 1 (FGF1) was detected by quantitative real-time polymerase chain reaction. Protein expression was checked by western blot. Cell viability, proliferation and apoptosis were investigated by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, 5-Ethynyl-29-deoxyuridine and flow cytometry analysis, respectively. Oxidative stress was evaluated by superoxide dismutase and catalase activity assay kits and lipid peroxidation malondialdehyde assay kit. Online databases and mechanism assays were used to predict and identify the relationship between miR-15a-5p and circ_0122396 or FGF1.

Results

Circ_0122396 and FGF1 expression were significantly downregulated, but miR-15a-5p expression was upregulated in ARC tissues or/and H₂O₂-treated SRA01/04 cells in comparison with control groups. H₂O₂ treatment repressed cell proliferation and induced cell apoptosis and oxidative stress, which was attenuated after circ_0122396 overexpression. MiR-15a-5p, a target mRNA of circ_0122396, was found to participate in H₂O₂-triggered cell damage by interacting with circ_0122396. Additionally, FGF1 silencing attenuated miR-15a-5p inhibitors-mediated action. Importantly, circ_0122396 regulated FGF1 expression by interaction with miR-15a-5p in H₂O₂-treated SRA01/04 cells.

Conclusion

Circ_0122396 ameliorated H₂O₂-triggered cell injury by inducing FGF1 through sponging miR-15a-5p, providing a potential target for ARC therapy.

中文翻译：

---

Circ_0122396 通过与 miR-15a-5p 结合刺激 FGF1 表达来保护人晶状体上皮细胞免受过氧化氢诱导的损伤

摘要

背景

据报道，Circular_0122396 (circ_0122396) 在年龄相关性白内障 (ARC) 中下调；然而，潜在的机制仍然未知。该研究旨在揭示 circ_0122396 在 ARC 进展及其机制中的作用。

方法

使用过氧化氢（H ₂ O ₂）诱导晶状体上皮细胞（SRA01/04）损伤。通过定量实时聚合酶链反应检测circ_0122396、microRNA-15a-5p（miR-15a-5p）和成纤维细胞生长因子1（FGF1）的RNA表达。通过蛋白质印迹检查蛋白质表达。分别通过 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide、5-Ethynyl-29-deoxyuridine 和流式细胞术分析研究细胞活力、增殖和凋亡。通过超氧化物歧化酶和过氧化氢酶活性测定试剂盒和脂质过氧化丙二醛测定试剂盒评估氧化应激。在线数据库和机制分析用于预测和识别 miR-15a-5p 与 circ_0122396 或 FGF1 之间的关系。

结果

_{与对照组相比，ARC组织或/和H 2} O ₂处理的SRA01/04细胞中Circ_0122396和FGF1表达显着下调，但miR-15a-5p表达上调。H ₂ O ₂处理抑制细胞增殖并诱导细胞凋亡和氧化应激，在circ_0122396过表达后减弱。MiR-15a-5p 是 circ_0122396 的靶 mRNA，被发现通过与 circ_0122396 相互作用参与 H ₂ O _{2触发的细胞损伤。}此外，FGF1 沉默减弱了 miR-15a-5p 抑制剂介导的作用。_{重要的是，circ_0122396 通过与 H 2} O ₂中的 miR-15a-5p 相互作用来调节 FGF1 的表达-处理过的 SRA01/04 细胞。

结论

Circ_0122396通过海绵化 miR-15a-5p 诱导 FGF1改善了 H ₂ O _{2触发的细胞损伤，为 ARC 治疗提供了潜在的靶点。}