抗VEGF是临床治疗视网膜黄斑变性(AMD)的主要手段，但许多患者对VEGF抗体治疗具有抗性。因此，寻找AMD对VEGF抗体治疗疗效不佳的原因具有重要的临床意义。C/ebp delta 是一种转录因子，在淋巴血管内皮细胞上，其调节VEGF-C自分泌信号通路影响淋巴血管的血管新生。我们的前期研究发现，缺氧促进内皮细胞的C/ebp delta表达；C/ebp delta的表达增高促进HIF-1和VEGF 的升高，因此提出如下假说：用VEGF抗体治疗AMD时，虽然阻断了外源性VEGF的作用，但由于缺氧引起 C/ebp delta表达增加，从而启动HIF-1-VEGF自分泌信号通路，降低VEGF抗体抑制血管新生的效果。我们拟通过细胞及动物实验、定量PCR、免疫印迹、基因敲除等方法研究C/ebp delta通过作用于VEGF的自分泌信号通路影响视网膜血管新生的作用及机制，为AMD的治疗提供新的途径。

VEGF antibody therapy is the main means of treatment to macular degeneration (AMD) in clinical practice. However，many patients are resistant to VEGF antibody therapy. Therefore, looking for molecular mechanisms of poor efficacy of VEGF antibody therapy to AMD has important clinical significance. C / ebp delta is a transcription factor, which was found to regulate VEGF-C autocrine signaling pathway on the lymphatic vessels of angiogenesis in the lymphatic endothelial cells. Our previous studies have found that hypoxia promotes C / ebp delta expression in endothelial cells; the increasing expression of C / ebp delta promotes expression of HIF-1 and VEGF, thus, we propose the following hypothesis: When treating AMD with VEGF antibody, even though exogenous VEGF paracrine effects on endothelial cell proliferation, migration and survival pathways are blocked , the hypoxia causes increasing expression of C / ebp delta, which initiates HIF-1- VEGF autocrine signaling pathway and reduces the effects of VEGF antibodies to inhibit angiogenesis. In this study, we intend to investigate how C / ebp delta affects the autocrine VEGF signaling and its effects on retinal vascular endothelial cell proliferation, migration, survival, and further study the mechanisms by the methods of cell culture, animal experiment, quantitative PCR, immunoblotting, gene knockout study etc; at mean time, we investigate whether it could enhance the effect of VEGF antibody treatment to AMD by the suppression of C / ebp delta. This would provide a new method for the treatment of AMD in clinical practice.

VEGF抗体治疗是近年来治疗视网膜黄斑变性(AMD)的主要手段，但许多患者对VEGF抗体治疗具有抗性。因此，寻找AMD对VEGF抗体治疗疗效不佳的分子机制具有重要的临床意义。C/ebp delta 是一种转录因子，研究发现在淋巴内皮细胞上，其调节VEGF-C自分泌信号通路影响淋巴血管的血管新生。我们通过细胞实验、动物实验、定量PCR、免疫印迹、基因敲除等方法研究C/ebp delta作用于血管内皮细胞VEGF的自分泌信号通路及机制。结果发现: 体外抑制血管内皮细胞的C/ebp delta可抑制血管内皮细胞的迁移和小管形成；体内实验结果显示在C/ebp delta基因敲除小鼠的主动脉负窗模型中，血管新生明显受到抑制。在小鼠的CNV模型中，经玻璃体腔内注射Lenti-C/ebp delta siRNA可抑制视网膜血管通透性升高和血管新生。其机制为，C/ebp delta和HIF-1的启动子区域结合，促进HIF-1升高；抑制血管内皮细胞的C/ebp delta通过HIF-1-VEGF 信号通路减少VEGF的产生，抑制 AMD的血管新生，从而为AMD的治疗提供新的途径。