EPO对糖尿病性视网膜前膜形成与发展的作用及机理研究

Diabetic retinopathy (DR) is one of the major diseases leading to blindness, and inhibiting formation and development of diabetic fibrovascular epiretinal membrane (dERM) is the key to save visual function for PDR patients. However, less studies exist about drugs for therapy of dERM, especially its underline mechanism is not completely understood. We found that erythropoietin (EPO) negatively regulated VEGF, and down-regulated expressions of IL-1β and TNF-α by Müller cells in diabetic retina, furthermore EPO could reduce Müller cell gliosis in diabetic retina. It is speculated that EPO may play a role in dERM formation and development mainly by suppressing pro-angiogenic factor, regulating cytoskeletal and cell biological behavior in Müller cells. The project establishes a novel co-culture model for dERM, and employs immunofluorescence, GST pull down, and cell scratch assays to study how EPO affects expression and release of pro-fibrous cytokines, the ratio of matrix metalloproteinases and their tissue inhibitors, and cytoskeleton and cell biological behavior changes in Müller cells. The obtaining data derived from the dERM model will result in explaining the role of EPO via Müller cells in the dERM formation and development, and indicating targets for the treatment of PDR.

糖尿病性视网膜病变（DR）是导致眼盲的重要疾病，抑制糖尿病性视网膜前纤维血管膜（dERM）形成与发展是挽救PDR患者视功能的关键。但目前干预治疗的药物研究较少，主要是作用机制尚未完全明确。我们发现促红细胞生成素（EPO）负反馈调节VEGF，还通过Müller细胞下调视网膜中IL-1β和TNF-α的表达，而且能减轻糖尿病视网膜胶质化反应。由此推测EPO可能主要通过调节Müller细胞中细胞因子表达，细胞骨架及细胞生物学行为改变等在dERM形成与发展中发挥作用。本项目建立模拟dERM的细胞模型，用免疫荧光、GST pull down、细胞划痕实验等方法研究EPO如何影响与dERM细胞共培养的Müller细胞中促纤维细胞因子的表达与释放，基质金属蛋白酶与其组织抑制剂的表达比例，细胞骨架及细胞生物学行为改变，从而明确EPO在dERM形成与发展中的作用及机理，为治疗DR提供依据和指导。

糖尿病性视网膜病变（简称“糖网病”）是全球性最主要的致盲眼病，牵拉性视网膜脱离是其致盲的主要原因之一，一旦发生其对视功能的损害往往是严重而不可逆的，因此早期抑制糖尿病性视网膜前纤维血管膜形成与发展是缓解病变进展的关键，然而目前缺少有效干预治疗方法。本项目证明了促红细胞生成素（EPO）能提高Müller细胞的活力，同时能抑制其在高糖环境下的增殖反应。此外，我们的研究还发现EPO能抑制转化生长因子-β（TGF-β）诱导下Müller细胞向成纤维细胞的表型转化、减少其产生促纤维化因子结缔组织生长因子（CTGF），并减少细胞金属蛋白酶9（MMP9）的表达。与此同时，我们还发现EPO能抑制Müller细胞的迁移，影响细胞骨架蛋白F-actin的构型。结果显示EPO通过提高Müller细胞自身活力，抑制增殖、转分化、迁移从而在糖尿病性视网膜前膜形成与发展过程中发挥保护作用。

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