高糖诱导中性粒细胞胞外陷阱（NETs）在糖尿病视网膜病变中的作用及机制研究

Diabetes mellitus is a group of metabolic disorders characterized by hyperglycemia, which leads to microvascular complications such as neuropathy,retinopathy and nephropathy. Diabetic retinopathy (DR) is the leading cause of blindness in the working age people worldwide, mainly because of the lack of effective treatment due to unknown pathogenesis of DR. In recent years, domestic and foreign researches have confirmed that para-inflammation as well as oxidative stress plays an important role in DR pathogenesis. There are more evidences indicate that neutrophils play a vital role in diabetes and its complications. Recently, people found another way of cell death in neutrophils called NETosis which is a form of neutrophil-specifc cell death characterized by the release of large web-like structures referred to as neutrophil extracellular traps (NETs). NETs are composed of DNA strands associated with histones and decorated with about 20 different proteins, including neutrophil elastase, myeloperoxidase, proteinase 3, high mobility group protein B1, and LL37. Recent data suggest that NETs plays a crucial role in the innate immune response and it is ROS-denpendent. In addtion, NETs do damage to vascular endothelial cells in vitro. Our latest study has found that high glucose can stimulate human neutrophils to form Neutrophil Extracellular Traps (NETs) . Thus, our present study intends to investigate the role and mechanism of NETs in diabetic retinopathy patients and streptozocin-induced type 2 diabetic rats.

糖尿病视网膜病变（DR）目前已成为影响全球性公共健康的主要疾病，是世界范围内致盲的主要原因，由于其发病机制尚不明确因而缺乏科学有效的防治手段。在DR的致病过程中，慢性低度炎症和氧化应激是其重要发病机制。近期研究发现中性粒细胞在激活后可释放出一种网状结构——中性粒细胞胞外陷阱（NETs）参与炎症反应。我们预实验发现中性粒细胞在高糖刺激下及DR患者血清刺激下较正常对照组产生NETs显著增加，基于以上前期基础，本项目以高糖诱导产生的NETs为切入点，以DR患者，DR动物模型为研究主体，以HRVEC为研究的靶标细胞，围绕氧化应激通路采用Real-time PCR、免疫荧光染色、ELISA、Western Blot、基因转染、流式细胞术等方法，从分子-细胞-动物-人群等不同层面着手，探讨NETs参与DR发病的具体作用及机制。

糖尿病视网膜病变（DR）目前已成为影响全球性公共健康的主要疾病，是世界范围内致盲的主要原因，由于其发病机制尚不明确因而缺乏科学有效的防治手段。在DR的致病过程中，慢性低度炎症和氧化应激是其重要发病机制。我们研究发现中性粒细胞在激活后可释放出一种网状结构——中性粒细胞胞外陷阱（NETs），并发现中性粒细胞在高糖刺激下及DR患者血清刺激下较正常对照组产生NETs显著增加。我们的数据表明，在高血糖环境下，中性粒细胞可发生网捕死亡产生NETs。高糖诱导NETs形成的具体机制与氧化应激机制有关。长期高血糖会导致血-视网膜屏障的破坏，为外周血中性粒细胞穿过血-视网膜屏障提供了更多的机会。越过屏障的中性粒细胞在高糖作用下产生NETs，沉积在玻璃体和视网膜中。同时，抗VEGF治疗可下调NETs的形成，这可能为DR的治疗提供新的靶点。