Liu Li-Li, Zhao Zhi-Yi
Department of Ophthalmology, Liuzhou Workers' Hospital, the Fourth Affiliated Hospital of Guangxi Medical University, Liuzhou 545005, Guangxi Zhuang Autonomous Region, China.
Int J Ophthalmol. 2023 Apr 18;16(4):514-522. doi: 10.18240/ijo.2023.04.04. eCollection 2023.
To investigate the retinoprotective role of Apigenin (Api) against high glucose (HG)-induced human retinal microvascular endothelial cells (HRMECs), and to explore its regulatory mechanism.
HRMECs were stimulated by HG for 48h to establish the cell model. Different concentrations of Api (2.5, 5, and 10 µmol/L) were applied for treatment. Cell counting kit-8 (CCK-8), Transwell, and tube formation assays were performed to examine the effects of Api on the viability, migration, and angiogenesis in HG-induced HRMECs. Vascular permeability was evaluated by Evans blue dye. The inflammatory cytokines and oxidative stress-related factors were measured using their commercial kits. Protein expression of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase 4 (NOX4) and p38 mitogen-activated protein kinase (MAPK) was measured by Western blot.
Api prevented HG-induced HRMECs viability, migration, angiogenesis, and vascular permeability in a concentration-dependent manner. Meanwhile, Api also concentration-dependently inhibited inflammation and oxidative stress in HRMECs exposed to HG. In addition, HG caused an elevated expression of NOX4, which was retarded by Api treatment. HG stimulation facilitated the activation of p38 MAPK signaling in HRMECs, and Api could weaken this activation partly downregulating NOX4 expression. Furthermore, overexpression of NOX4 or activation of p38 MAPK signaling greatly weakened the protective role of Api against HG-stimulated HRMECs.
Api might exert a beneficial role in HG-stimulated HRMECs through regulating NOX4/p38 MAPK pathway.
研究芹菜素（Api）对高糖（HG）诱导的人视网膜微血管内皮细胞（HRMECs）的视网膜保护作用，并探讨其调节机制。
用HG刺激HRMECs 48小时以建立细胞模型。应用不同浓度的Api（2.5、5和10μmol/L）进行处理。进行细胞计数试剂盒-8（CCK-8）、Transwell和管形成试验，以检测Api对HG诱导的HRMECs活力、迁移和血管生成的影响。用伊文思蓝染料评估血管通透性。使用其商业试剂盒测量炎性细胞因子和氧化应激相关因子。通过蛋白质免疫印迹法测量烟酰胺腺嘌呤二核苷酸磷酸（NADPH）氧化酶4（NOX4）和p38丝裂原活化蛋白激酶（MAPK）的蛋白表达。
Api以浓度依赖的方式阻止了HG诱导的HRMECs活力、迁移、血管生成和血管通透性。同时，Api还以浓度依赖的方式抑制了暴露于HG的HRMECs中的炎症和氧化应激。此外，HG导致NOX4表达升高，而Api处理可使其降低。HG刺激促进了HRMECs中p38 MAPK信号的激活，而Api可通过下调NOX4表达部分减弱这种激活。此外，NOX4的过表达或p38 MAPK信号的激活大大削弱了Api对HG刺激的HRMECs的保护作用。
Api可能通过调节NOX4/p38 MAPK途径对HG刺激的HRMECs发挥有益作用。
