NLRP3介导炎症反应参与糖尿病视网膜病变的作用机制及其相关基因多态性的功能研究

The pathogenesis of diabetic retinopathy (DR) is believed to be multifactorial, with inflammation and genetic factors playing a fundamental role. NLRP3 is one of the best-studied members of the nucleotide-binding oligomerizatin domain (NOD)-like receptor (NLR) family. The dysregulation of NLRP3 inflammasome activity has been demonstrated to lead numerous proinflammatory, non-microbial human diseases. NLRP3 gene mutation is also associated with autoinflammatory diseases, metabolic disorders, and so on. We found that the higher mRNA expression level of NLRP3 in peripheral blood mononuclear cells from patients with DR. Therefore, we speculate that NLRP3 plays an important role in DR. On that basis, in the current study, expression and activation of NLRP3 inflammasome will be observed, the mechanism of mitochondrial reactive oxygen species required for NLRP3 inflammasome activation will be clarified, and the influence of local treatment of DR drugs on NLRP3 inflammasome will be determined in patients with DR and diabetic mouse. Meanwhile, using our database and blood specimen bank about patients from type 2 diabetes and DR, which have been set up completely through epidemiological investigation about DR based on inpatients and community residents, the polymorphisms of NLRP3 gene will be examined to assess their contribution to DR, and the effects of relative polymorphisms on NLRP3 expressions will be analyzed. This study will lay the theoretical basis for etiology and pathogenesis of DR, and will provide a new molecular target for individualized prevention and treatment of DR.

炎症反应与遗传因素已被证实在糖尿病视网膜病变（DR）中具有重要作用。NLRP3作为NOD样受体家族的重要成员，其炎症体通过介导免疫应答加强机体的炎症反应，其基因突变与代谢紊乱、慢性炎症等多种疾病密切相关。目前我们发现NLRP3在DR患者中存在异常表达，据此推测NLRP3可能通过介导炎症反应参与DR的发生。为验证该假说，本项研究将以NLRP3为切入点，以DR患者及糖尿病小鼠为研究对象，进一步明确NLRP3及其炎症体在DR中的表达与活化特点，阐明线粒体活性氧诱导其活化的机制，确定DR药物局部治疗对NLRP3炎症体的影响；同时将利用我们已建立的2型糖尿病及DR患者临床资料和血液标本库，明确NLRP3基因多态性与DR的相关性，揭示其相关多态参与DR发生的生物学功能。上述研究旨在明确NLRP3参与DR的作用机制，进而为阐明DR的病因和发病机制奠定理论基础，并将为DR的个体化防治研究提供新的切入点。

炎症反应与遗传因素已被证实在糖尿病视网膜病变中具有重要作用。NLRP3作为NOD样受体家族的重要成员，其炎症体通过介导免疫应答加强机体的炎症反应，其基因突变与代谢紊乱、慢性炎症等多种疾病密切相关。本项目研究发现NLRP3在糖尿病视网膜病变患者外周血单个核细胞中表达水平显著升高，并进一步证实了高糖环境可激活视网膜血管内皮细胞发生炎症反应，活化NLRP3炎症小体，进而导致视网膜血管内皮细胞增殖，同时加速细胞凋亡。虽然有研究证实NLRP3基因多态性与糖尿病、肥胖、自身免疫性疾病、慢性炎症等疾病的发生密切相关，但针对全外显子测序的结果显示，NLRP3多态性与糖尿病视网膜病变的发生没有相关性，但某些单倍体型在糖尿病视网膜病变组和非糖尿病视网膜病变组中表现出了较显著的差异。项目执行过程中进一步健全扩充了糖尿病视网膜病变患者标本库，总结报道了南方人群糖尿病患者低视力和盲的患病率及主要致盲原因，以及该人群糖尿病视网膜病变的患病率和危险因素，丰富了国人相关数据信息，为糖尿病人群的相关眼病群体研究提供有力的数据及借鉴。本项目的开展明确了NLRP3参与糖尿病视网膜病变的作用机制，进而为阐明该病的病因和发病机制奠定理论基础，并将为个体化防治研究提供新的切入点。