丁苯酞通过Nrf2通路调控糖尿病视网膜病中视网膜Müller细胞NLRP3炎症小体介导的焦亡的作用及机制研究

Diabetic retinopathy (DR) is an inflammatory disease. Pyrotosis is highly inflammatory-relevant programmed cell death. NLRP3 inflammasome is a vital regulator for pyrotosis, and NLRP3 inflammasome-mediated pyrotosis of the retinal Müller cells is a potential therapeutic target for DR treatment. As a natural Nrf2 activator, 3-N-butylphthalide (NBP) can regulate NLRP3 expression and possess anti-diabetic, anti-inflammatory and neuroprotective roles. However, the effect of NBP on NLRP3 inflammasome expression of retinal Müller cells in DR has not been studied yet. Our preliminary experiment showed that NBP significantly decreased high glucose-induced NLRP3 protein expression in cultured retinal Müller cells. In this study, NBP was treated to the cultured retinal Müller cells under high glucose. STZ diabetic mice models were established and NBP were given to both wild-type and Nrf2-/- mice by intragastric administration. Western blot, RT-PCR, ELISA, immunofluorescence staining, transmission electron microscope scanning were applied to explore the effect and the mechanism of NBP, both in vitro and in vivo, on NLRP3 inflammasome-mediated the pyrotosis of the retinal Müller cells in DR. This study may lay the theoretical foundation for exploring NBP as a promising therapeutic medication for DR treatment.

糖尿病视网膜病变（DR）是一种慢性炎症性疾病。焦亡是与炎症高度相关的程序性细胞死亡模式。NLRP3炎症小体是焦亡的重要调控因子，其介导的视网膜Müller细胞焦亡是DR干预的新靶点。丁苯酞（NBP）是重要的抗氧化因子Nrf2的天然激动剂，能调控NLRP3表达，具有抗炎、抗糖尿病和神经保护作用。但NBP对DR以及对视网膜Müller细胞NLRP3的作用及调控机制尚不明确。本课题组预实验表明NBP能降低高糖诱导的Müller细胞NLRP3蛋白的表达。因此，本课题拟通过对体外培养高糖条件下的Müller细胞进行NBP干预，并建立野生型和Nrf2-/-糖尿病小鼠模型，给予NBP灌胃，应用western blot、免疫荧光等技术，从体外和体内水平深入研究NBP通过Nrf2通路调控DR中Müller细胞NLRP3炎症小体介导的焦亡的作用及其机制，为NBP可能成为治疗DR的新药提供理论依据。

糖尿病视网膜病变（DR）是一种慢性炎症性疾病。焦亡是与炎症高度相关的程序性细胞死亡模式。NLRP3炎症小体是焦亡的重要调控因子，其介导的视网膜Müller细胞焦亡是DR干预的新靶点。丁苯酞（NBP）是重要的抗氧化因子Nrf2的天然激动剂，能调控NLRP3表达，具有抗炎、抗糖尿病和神经保护作用。但NBP对DR以及对视网膜Müller细胞NLRP3的作用及调控机制尚不明确。本课题组通过对体外培养高糖条件下的Müller细胞进行NBP干预，结果表明NBP能够抑制高糖诱导的Müller细胞的氧化应激ROS、炎症（TNFa、IL-1β、NLRP3、pro-caspase 1），促进抗氧化通路（Nrf2）的激活，深入阐述了NBP通过激活Nrf2通路调控DR中Müller细胞NLRP3炎症小体介导的焦亡的作用及其机制.此外，NBP还能降低高糖诱导的Müller细胞的增殖、迁移、抑制VEGF以及内皮细胞血管生成。综上，NBP对高糖刺激的Müller细胞具有抗炎症、抗氧化、抗增殖、抗迁移、抗焦亡的作用，对血管内皮细胞具有抗血管新生的作用，对糖尿病视网膜的发生发展具有保护作用，NBP可能成为糖尿病视网膜病治疗的潜在新靶点。