脯氨酰羟化酶2（PHD2）与糖尿病视网膜病变新生血管形成的关系及作用机制研究

Diabetic retinopathy (DR) is the most common and most severe diabetic microangiopathy, and take the first place in blinding retinal disease. Neovascularization is the main pathological manifestation of DR, but its mechanism is still unknown, and no ideal treatment method. Approaching to the occurrence and control mechanisms contribute to the elucidation of the pathogenesis of DR, providing potential target for the treatment of early DR. The PHD2 is a key molecules adjusting HIF-1a and related with tumor angiogenesis, and also as an important regulatory factor take part in cell proliferation. Our preliminary experiment found the expression of PHD2 was positive in intraocular fibrovascular membrane from patients with the proliferative DR. In vitro, cultured vascular endothelial cells also showed the positive expression of PHD2, and the mRNA expression of PHD2 was increased in retina of early diabetic rat, which may indicate that PHD2 take part in DR neovascularization and play a key role. In this research, application of small interfering RNA technology, at the cellular and animal level silencing of PHD2 gene expression, we verificate the relationship between PHD2 and angiogenic factor, clarify angiogenesis mechanism effect by PHD2, and find its interaction site in vascular regulatory networks and evaluate its therapeutic efficacy.

糖尿病视网膜病变（Diabetic retinopahty,DR）是糖尿病最常见和最严重的微血管病变，在致盲性视网膜血管疾病中排名第一。新生血管形成是DR 发生的主要病理表现，但其机制不明，亦无理想治疗方法。对其发生和调控机制的探讨有助于阐明DR 的发病机制，为早期治疗提供潜在靶点。PHD2是调节HIF-1a的关键分子，参与肿瘤新生血管形成，也是细胞迁移、增殖重要的调控因子。我们预实验发现其在增殖性DR 患者眼内纤维血管膜上呈阳性表达，在体外培养的血管内皮细胞中亦呈阳性表达，在早期糖尿病大鼠视网膜中mRNA表达增高，这些都提示PHD2可能参与了DR新生血管的形成并起关键作用。本课题拟应用RNA干扰技术，在细胞和动物水平沉默PHD2表达，验证其与血管生成调节因子之间的功能联系以及可能在血管调控网络中的作用位点，初步阐明PHD2在DR中调控血管形成的作用机理，评价其作为靶向治疗药物的有效性。

糖尿病视网膜病变已成为我国主要的致盲眼病之一，对其发现和早期治疗可以挽救患者视力，提高生存质量。本实验主要研究糖尿病视网膜病变发病过程中可能起关键作用的因子PHD2，通过从细胞和动物水平验证PHD2在视网膜新生血管性疾病中的与新生血管和细胞增殖相关的功能作用，并分析其可能的作用机制，评价抑制或敲减PHD2之后，对视网膜新生血管发生发展的治疗效果。同时，在本实验中观察和研究PHD2与GFAP，VEGF等其它血管新生相关细胞因子之间的联系，并且在实验进展过程中，发现PHD2和视网膜神经胶质细胞、视网膜色素上皮细胞、微血管内皮细胞等均具有相关性，PHD2在糖尿病视网膜病变的发生发展中具有重要的作用。