Department of Sense Organs, University of Rome Sapienza, Rome, Italy.
National Research Council - Department of Biomedical Sciences, Institute of Cell Biology and Neurobiology (IBCN-CNR), Rome, Italy.
J Cell Physiol. 2018 Nov;233(11):8874-8883. doi: 10.1002/jcp.26806. Epub 2018 Jun 1.
Intravitreal injection (IVT) of antivascular endothelial growth factor (anti-VEGF) agents is widely used for the treatment of retinal vascular diseases. Recently, the injection of anti-VEGF agents in the ocular anterior chamber has been proposed for the treatment of neovascular glaucoma and potential side effects on the corneal structures have been investigated with contrasting results. Increasing evidence has demonstrated that VEGF inhibition is associated with cellular apoptotic changes and that this effect may be mediated by alterations in nerve growth factor (NGF) pathway. In this study, we demonstrated that anterior chamber injection (IC), but not IVT injection of two different anti-VEGF agents, aflibercept and ranibizumab, affects rabbit corneal endothelium in terms of survival and apoptosis and is associated with changes in endothelial expression of NGF precursor (proNGF) and p75 neurotrophin receptor (p75NTR) receptor. We observed an increase in corneal endothelial cell incorporation of trypan blue and expression of cleaved-caspase 3 (c-Casp3), p75NTR, and RhoA after IC injection of both anti-VEGF drugs when compared with the vehicle. Our results showed that apoptosis induction by aflibercept was more pronounced when compared with that of ranibizumab. Aflibercept also mediated a significant increase in endothelial expression of proNGF when compared with the vehicle. In line with these data, IC administration of both anti-VEGF agents induced the activation of apoptotic signals in endothelial cells, including an increase in c-Casp3, decrease in Bad Ser 112 phosphorylation, and unbalance of AKT phosphorylation. These results demonstrated that administration of anti-VEGF in the anterior chamber of rabbit affects endothelial cell survival by inducing apoptosis through alteration of NGF pathway.
玻璃体内注射（IVT）抗血管内皮生长因子（anti-VEGF）药物被广泛用于治疗视网膜血管疾病。最近，已经提出在前房内注射抗 VEGF 药物来治疗新生血管性青光眼，并且已经用对比结果研究了对角膜结构的潜在副作用。越来越多的证据表明，VEGF 抑制与细胞凋亡变化有关，并且这种作用可能是通过神经生长因子（NGF）途径的改变介导的。在这项研究中，我们证明了前房内注射（IC），而不是 IVT 注射两种不同的抗 VEGF 药物，即阿柏西普和雷珠单抗，会影响兔子角膜内皮细胞的存活和凋亡，并与内皮细胞表达 NGF 前体（proNGF）和 p75 神经营养因子受体（p75NTR）受体的改变有关。与载体相比，我们观察到在 IC 注射两种抗 VEGF 药物后，角膜内皮细胞对台盼蓝的摄取和裂解半胱天冬酶 3（c-Casp3）、p75NTR 和 RhoA 的表达增加。与雷珠单抗相比，阿柏西普诱导的细胞凋亡更为明显。与载体相比，阿柏西普还介导了内皮细胞 proNGF 表达的显著增加。与这些数据一致，两种抗 VEGF 药物的 IC 给药均诱导内皮细胞中凋亡信号的激活，包括 c-Casp3 增加、Bad Ser 112 磷酸化减少和 AKT 磷酸化失衡。这些结果表明，抗 VEGF 在兔子前房内的给药通过改变 NGF 途径诱导内皮细胞凋亡而影响内皮细胞的存活。
