Transforming Growth Factor Beta-Activated Kinase 1 (Tak1) in Retinal Microglial Inflammation

糖尿病视网膜病变与慢性异常炎症有关，据认为，慢性异常炎症在糖尿病视网膜病变中发挥着关键作用。 神经血管单元功能的早期破坏。关于细胞内信号传导的信息不完整 参与炎症信号上调的途径。识别更多途径成员将导致 更多的治疗靶点可用于治疗神经退行性疾病。该提案旨在 测试转化生长因子 β 激活激酶 1 (TAK1) 在激活 NFκB、p38 和 JNK 中的作用 视网膜小胶质细胞中的通路，导致炎症因子转录增加。假设 抑制 TAK1 将减少糖尿病视网膜病变早期阶段的炎症，并减少糖尿病视网膜病变的变化 神经血管单元将使用分离的小胶质细胞进行体外测试，并使用以下组合进行体内测试 链脲佐菌素 (STZ) 在小胶质细胞中条件性丧失 TAK1 的小鼠中诱导糖尿病。实验将 利用小分子抑制剂，添加已知的炎症因子，显微注射，定量 聚合酶链反应、蛋白质印迹分析、多重酶联免疫吸附测定以及 视网膜中周细胞、微血管、小胶质细胞、视网膜星形胶质细胞、神经节细胞和穆勒胶质细胞的定量 整体和部分。

Diabetic retinopathy is associated with chronic aberrant inflammation that is proposed to play a critical role in the early disruption of neurovascular unit function. There is incomplete information about intracellular signaling pathways involved in upregulation of inflammatory signals. Identification of more pathway members will lead to more therapeutic targets to be used in treatment of neurodegenerative diseases. This proposal is aimed at testing the role of transforming growth factor β-activated kinase 1 (TAK1) in activation NFκB, p38, and JNK pathways in retinal microglia, resulting in increased transcription of inflammatory factors. The hypothesis that inhibition of TAK1 will reduce inflammation in the early stages of diabetic retinopathy and reduce changes to the neurovascular unit will be tested in vitro using isolated microglial cells and in vivo using a combination of streptozotocin (STZ)-induced diabetes in mice with conditional loss of TAK1 in microglia. Experiments will utilize small molecule inhibitors, addition of known inflammatory factors, microinjections, quantitative polymerase chain reactions, Western blot analysis, multiplex enzyme-linked immunosorbent assays, and quantitation of pericytes, microvasculature, microglia, retinal astrocytes, ganglion cells, and Müller glia in retinal wholemounts and sections.