小分子化合物通过抑制GSK-3β活性与Notch信号调控大鼠视网膜Müller细胞重编程的作用研究

Our previous studies have confirmed that mammalian Müller cells can proliferate in retinal lesions and even reprogram the progenitor cells and regenerate neurons. However, this regeneration capacity is very limited and can not be sustained, it is difficult to achieve the purpose of effective repair. Therefore, in the process of Müller reprogramming how to regulate and promote the proliferation of cells more differentiation into the structure and function of mature neurons, as retinal damage and repair to be overcome scientific problems. On the basis of literature review and pre-experiment, we proposed the two important signaling pathways Wnt and Notch in the process of Müller's reprogramming process. By using retinal degeneration rat model and in vitro cell culture pathway, we used single or combined small molecule Inhibitory effects of Wnt and Notch on Müller cell reprogramming and neuronal regeneration were investigated by investigating these two signaling pathways, and the molecular mechanism was explored in order to achieve the mechanism of reprogramming Müller Cells are more effectively transdifferentiated into retinal structures and functional neurons for the purpose of repairing the clinical retinal degeneration of endogenous regeneration therapy to provide a theoretical basis and experimental basis.

我们前期研究证实哺乳类动物Müller细胞在视网膜病变环境下能进行大量增殖，甚至具有重编程为祖细胞，并再生神经元的潜能。然而，这种再生能力却非常有限和不能持久，难以到达有效修复的目的。因此，在Müller重编程过程中如何调控和促进增殖的细胞更多地分化为结构和功能成熟的神经元，成为视网膜损伤与修复有待攻克的科学问题。我们在文献综述和预实验的基础上，提出以Müller重编程过程中两个重要的信号通路Wnt和Notch为靶点，通过视网膜变性大鼠模型和体外细胞培养途径，采用单独或联合应用小分子化合物抑制剂GSK-3βI和DAPT，分别对这两个信号通路进行干预，进一步揭示Wnt和Notch在Müller细胞重编程和神经元再生过程中的作用规律，探索其分子机制，以实现把重编程Müller细胞更有效地转分化为视网膜结构与功能修复所需的神经元为目的，为临床视网膜变性疾病的内源性再生治疗提供理论基础和实验依据。

我们前期研究证实哺乳类动物Müller细胞在视网膜病变环境下能进行增殖，甚至具有重编程为祖细胞，并再生神经元的潜能。然而，这种再生能力却非常有限和不能持久，难以达到有效修复的目的。因此，如何调控Müller重编程过程，以促进更多地分化成新神经元，成为视网膜损伤与修复有待攻克的科学问题。我们在文献综述和预实验的基础上，提出以Müller重编程过程中两个重要的信号通路Wnt和Notch为靶点。首先证实GSK-3β在成年大鼠的分布和细胞定位；然后通过视网膜变性大鼠模型，采用单独或联合应用小分子化合物抑制剂GSK-3βI，观察其对Müller重编程过程的作用。 结果发现，GSK-3βI上调Nestin表达，伴随着MG增殖不变，反而促进小胶质细胞发生非经典活化和增殖。联合应用GSK-3βI与DAPT对修复期新生神经元的作用也不大。然而，我们发现GSK-3βI具有明显的神经保护作用，能有效减少受损神经元数量，从而达到视网膜结构与功能修复的目的，为临床视网膜变性疾病的治疗提供理论基础和实验依据。