靶向髓样分化蛋白2的小分子抑制剂对视网膜缺血再灌注损伤的疗效研究

Inflammation and Oxidative stress plays an important role in the progress and development of Retinal Ischemia-Reperfusion (RIR) Injury. TLR4/MD2 is an important mediating receptor for LPS-induced inflammatory response. However, it is unclear whether MD2 plays a role in RIR injury in which LPS is not involved. In the previous studies on anti-inflammatory drug design and evluation, we found a novel molecule, L2H17, which directly targeted MD2 protein and possessed excellent anti-inflammatory activity in LPS-stimulated inflammatory models. Interestingly, oral administration of compound L2H17 could significantly attenuate retina injury in RIR rat models. In view of these results, we hypothesize here, that MD2 may play an important role in the inflammation and oxidation stress involved in RIR injury, and inhibition of MD2 may contribute to the treatment of DR. A series of studies are designed to test this hypothesis. These include, 1) to investigate the TLR4/MD2-dependent mechanism by which L2H17 inhibits TBHP-induced inflammation in retinal pigment epithelial cells and macrophages, 2) to probe the MD2-mediated pharmacology of L2H17 in RIR mice, 3) to identify the endogenous ligand of MD2 in TBHP-induced inflammation and RIR by the method of proteomics, 4)and to demonstrate the important role of MD2 in RIR injury using MD2 gene knockout mice. There is an urgent demand to develop more reliable drug target and treatment in RIR. With the completion of this challenge project, we will demonstrated the MD2-involved mechanism in RIR injury and provide a new therapeutic target for the treatment of RIR.

炎症和氧化应激是视网膜缺血再灌注（RIR）损伤的主要介导过程。TLR4/MD2是介导内毒素LPS炎症的重要受体复合物。TLR4在RIR损伤中有着重要作用，但MD2是否参与非LPS条件下的RIR损伤过程尚未可知。项目组前期工作寻找到特异性的MD2抑制剂L2H17，它具有优秀抗炎活性，且在体内显著缓解大鼠RIR损伤。基于此，我们假设：MD2在RIR损伤过程中有着重要的介导作用，可以成为新的治疗靶点。本项目拟利用多种细胞、MD2基因沉默小鼠等，探讨MD2在体外TBHP刺激和体内RIR损伤的过程中介导炎症和氧化应激的作用，及其在L2H17抑制RIR损伤中的介导作用机制；最后利用蛋白质组学寻找TBHP刺激和RIR损伤对激活MD2的内源性配体的影响，阐明MD2的上游机制。项目将深入阐明L2H17防治RIR损伤的靶点和药理药效，明确MD2在RIR损伤中的作用，证实其可以作为相关眼科疾病防治的新靶点。

炎症和氧化应激是视网膜缺血再灌注（RIR）损伤的主要介导过程。TLR4/MD2是介导内毒素LPS 炎症的重要受体复合物。TLR4 在RIR 损伤中有着重要作用，但MD2 是否参与非LPS 条件下的RIR 损伤过程尚未可知。项目组前期工作寻找到特异性的MD2 抑制剂L2H17，它具有优秀抗炎活性，且在体内显著缓解大鼠RIR 损伤。基于此，我们假设：MD2在RIR 损伤过程中有着重要的介导作用，可以成为新的治疗靶点。本项目利用多种细胞、MD2 基因沉默小鼠等，探讨MD2 在体外TBHP 刺激和体内RIR 损伤的过程中介导炎症和氧化应激的作用，及其在L2H17 抑制RIR 损伤中的介导作用机制；最后利用蛋白质组学寻找TBHP刺激和RIR 损伤对激活MD2 的内源性配体的影响，阐明MD2 的上游机制。项目深入阐明L2H17 防治RIR 损伤的靶点和药理药效，明确MD2 在RIR 损伤中的作用，证实其可以作为相关眼科疾病防治的新靶点。