Abstract：
Corneal neovascularization (CNV) is a vision-threatening disease that is becoming a growing public health concern. While Yes-associated protein (YAP) plays a critical role in neovascular disease and allow for the sprouting angiogenesis. Verteporfin (VP) is a classical inhibitor of the YAP-TEAD complex, which is used for clinical treatment of neovascular macular degeneration through photodynamic therapy. The purpose of this study is to explore the effect of verteporfin (VP) on the inhibition of CNV and its potential mechanism. Rat CNV model were established by suturing in the central cornea and randomly divided into three groups (control, CNV and VP group). Neovascularization was observed by slit lamp to extend along the corneal limbus to the suture line. RNA-sequencing was used to reveal the related pathways on the CNV and the results revealed the vasculature development process and genes related with angiogenesis in CNV. In CNV group, we detected the nuclear translocation of YAP and the expression of CD31 in corneal neovascular endothelial cells through immunofluorescence. After the application of VP, the proliferation, migration and the tube formation of HUVECs were significantly inhibited. Furthermore, VP showed the CNV inhibition by tail vein injection without photoactivation. Then we found that the expression of phosphorylated YAP significantly decreased, and its downstream target protein connective tissue growth factor (CTGF) increased in the CNV group, while the expression was just opposite in other groups. Besides, both the expression of vascular endothelial growth factor receptor 2 (VEGFR2) and cofilin significantly increased in CNV group, and decreased after VP treatment. Therefore, we conclude that Verteporfin could significantly inhibited the CNV without photoactivation by regulating the activation of YAP.
摘要：
角膜新生血管形成（CNV）是一种威胁视力的疾病，正成为越来越多的公共卫生问题。而Yes相关蛋白（YAP）在新生血管疾病中起着关键作用，并允许发芽血管生成。Verteporfin（VP）是YAP-TEAD复合物的经典抑制剂，光动力疗法用于新生血管性黄斑变性的临床治疗。本研究旨在探讨维替泊芬(VP)对CNV的抑制作用及其潜在机制。采用中央角膜缝合法建立大鼠CNV模型,随机分为3组(对照组,CNV和VP组)。通过裂隙灯观察到新血管形成沿角膜缘延伸至缝合线。RNA测序用于揭示CNV上的相关途径，结果揭示了CNV中血管发育过程和与血管生成相关的基因。在CNV组中，我们通过免疫荧光检测了角膜新生血管内皮细胞中YAP的核易位和CD31的表达。申请VP后，扩散,显著抑制HUVECs的迁移和管腔形成。此外，VP显示通过尾静脉注射而没有光活化的CNV抑制。然后我们发现磷酸化YAP的表达显著降低,其下游靶蛋白结缔组织生长因子（CTGF）在CNV组中增加，而其他组的表达正好相反。此外,CNV组血管内皮生长因子受体2(VEGFR2)和cofilin的表达均显著增加,VP治疗后下降。因此,我们得出结论，Verteporfin可以通过调节YAP的激活而显着抑制CNV而没有光活化。
