Eye Institute of Xiamen University, Medical College of Xiamen University, Fujian Provincial Key Laboratory of Ophthalmology and Visual Science, Xiamen, Fujian 361102, P.R. China.
Department of Ophthalmology, The First Affiliated Hospital of Nanchang University, Nanchang, Jiangxi 330006, P.R. China.
Mol Med Rep. 2018 Feb;17(2):2187-2194. doi: 10.3892/mmr.2017.8114. Epub 2017 Nov 20.
To investigate the effects of catalpol on corneal neovascularization (CNV) and associated inflammation, eye drops (5 mM catalpol or PBS) were administered four times daily to alkali‑burn rat models of CNV and inflammation. Clinical evaluations of CNV and the degree of inflammation were performed on days 0, 4, 7, 10 and 14 under slit lamp microscopy. Eyes were collected on day 14 and prepared for hematoxylin and eosin, and immunofluorescence staining; corneal cell apoptosis was investigated via terminal deoxynucleotidyl transferase‑mediated nick end labeling (TUNEL) staining. Protein expression levels of angiogenic and proinflammatory factors, including vascular endothelial growth factor (VEGF), pigment epithelium‑derived factor (PEDF), tumor necrosis factor‑α (TNF‑α) and necrosis factor‑κB (NF‑κB) were determined by western blotting. The effects of catalpol on cell proliferation were investigated in vitro using human umbilical vein endothelial cells (HUVECs) and a Cell Counting kit‑8 (CCK‑8); alterations in migration and tube formation were investigated via HUVEC wound closure and tube formation assays. HUVEC viability and proliferative ability were inhibited in a dose‑dependent manner; catalpol also decreased HUVEC cell migration and tube forming ability. Within alkali‑burn rat models, decreased inflammation and CNV was associated with catalpol administration; as demonstrated with TUNEL, corneal cell apoptosis was decreased in response to catalpol. Western blot analysis revealed reduced protein expression levels of VEGF and TNF‑α; however, PEDF and phosphorylated‑NF‑κB p65 were increased due to catalpol administration. The present study demonstrated the inhibitory effects exerted by catalpol on CNV and inflammation within alkali‑burned rat models. Topical application of catalpol in vivo was associated with reduced CNV and inflammation; therefore, catalpol may be considered an anti‑inflammatory agent for the clinical treatment of CNV.
为了研究梓醇对角膜新生血管（CNV）和相关炎症的影响，将眼滴（5 mM 梓醇或 PBS）每天四次施用于碱烧伤大鼠 CNV 和炎症模型。在裂隙灯显微镜下于 0、4、7、10 和 14 天对 CNV 和炎症程度进行临床评估。在第 14 天收集眼睛，进行苏木精和伊红、免疫荧光染色；通过末端脱氧核苷酸转移酶介导的缺口末端标记（TUNEL）染色研究角膜细胞凋亡。通过蛋白质印迹法测定血管生成和促炎因子，包括血管内皮生长因子（VEGF）、色素上皮衍生因子（PEDF）、肿瘤坏死因子-α（TNF-α）和核因子-κB（NF-κB）的蛋白表达水平。使用人脐静脉内皮细胞（HUVEC）和细胞计数试剂盒-8（CCK-8）在体外研究梓醇对细胞增殖的影响；通过 HUVEC 伤口闭合和管形成测定研究迁移和管形成的变化。HUVEC 活力和增殖能力呈剂量依赖性抑制；梓醇还降低了 HUVEC 细胞迁移和管形成能力。在碱烧伤大鼠模型中，炎症和 CNV 的减少与梓醇的给药有关；TUNEL 显示，梓醇处理后角膜细胞凋亡减少。Western blot 分析显示 VEGF 和 TNF-α 的蛋白表达水平降低；然而，由于梓醇的给药，PEDF 和磷酸化-NF-κB p65 增加。本研究表明梓醇对碱烧伤大鼠模型中 CNV 和炎症具有抑制作用。体内局部应用梓醇与减少 CNV 和炎症有关；因此，梓醇可被视为治疗 CNV 的临床抗炎药。
