NLRP3炎症小体介导的干眼氧化损伤所致的炎症机制研究

干眼是眼科门诊患者中的第二大眼病，目前认为其核心发病机制是泪液高渗和免疫炎症反应，然而干眼患者眼表组织中高渗引起的氧化损伤也日益被重视，但这三者在干眼发病过程中的相互关系尚不明确。最近研究发现，细胞氧化损伤中过多的ROS，通过激活NLRP3炎症小体，而引发炎症反应。据此，我们认为干眼可能是由于泪液高渗产生的ROS激活眼表的NLRP3炎症小体进而激发炎症反应，最终导致干眼发病。本课题利用单纯环境因素诱导的小鼠干眼模型，研究其眼表的ROS、炎症小体（NLRP3、ASC、Caspase1）和炎症因子（IL-1β、IL-6、IL-18 等）的表达，并通过氧化损伤抑制剂反向验证；其次，体外培养角膜和结膜上皮细胞，观察其ROS、NLRP3炎症小体和炎症因子对高渗透压的反应，并通过上调和下调NLRP3的表达，深入研究其在干眼发病中的机制。

Dry eye syndrome (DES), the second top eye disease in the outpatients, is a multifactorial disease with potential damage to the ocular surface. It is accompanied by increased osmolarity of the tear film and inflammation of the ocular surface, and the inflammation is the most accepted mechanism of DES, while the oxidative stress is also discovered to be related to DES, It has been reported that excessive ROS stimulates NLRP3 to form NLRP3 inflammasome, which activates the maturation and secretion of IL-1β and IL-18 cytokines. And we presume that this is the way how oxidative stress triggers inflammation in DES. This study uses a dry eye mouse model induced by single environmental factor, which was established in our lab, to investigate the expression of ROS, NLRP3 inflammasome components (NLRP3, ASC, Caspase1), and inflammatory factors (IL-1β, IL-6, and IL-18) in mouse ocular surface with or without the use of ROS inhibitor N-acetyl-L-cysteine (NAC). And the expressions of these factors are also investigated in cultured human permanent corneal epithelial cells and primary conjunctival epithelial cells in high-osmolality media. Then the treatments of knock-down and knock-up of NLRP3 in these cells are performed to validate the effect of NLRP3 system in the development of DES.