大鼠视网膜Müller细胞在高糖、氧化应激、缺氧中的表现

The characteristics of rat retinal Müller cells in high glucose, oxidative stress and hypoxia

目的观察Sprague-Dawley(SD)大鼠视网膜Müller细胞在高糖、氧化应激、缺氧三种病理环境下的特征,为糖尿病视网膜病变的防治提供新的研究依据。方法体外培养SD大鼠视网膜Müller细胞,并用谷氨酰胺合成酶(GS)鉴定。将传2代的细胞随机分为四组:对照组、高糖组、氧化应激组、缺氧组。倒置显微镜观察细胞形态学改变,免疫荧光染色法观察细胞GS和α-平滑肌肌动蛋白(α-SMA)的变化,Transwell小室观察细胞迁移能力。结果体外成功培养并鉴定鼠Müller细胞,四组Müller细胞功能均受损,氧化应激组以及缺氧组结构受损明显,失去正常形态。缺氧组中α-SMA的表达呈阳性,并且在Transwell小室中发生迁移的细胞数明显多于其他组,差异有统计学意义(P < 0.05)。结论高糖、氧化应激、缺氧均可导致Müller细胞功能受损,缺氧可诱导Müller细胞转化为具有迁移能力的肌纤维样细胞,提示Müller细胞参与了增生性糖尿病视网膜病变的发病过程。

Objective To observe the characteristics of Sprague-Dawley (SD) rats retinal Müller cells under three pathological conditions including high glucose, oxidative stress and hypoxia, and to provide new research basis for the prevention and treatment of diabetic retinopathy. Methods SD rat retinal Müller cells were cultured in vitro and identified by glutathione synthetase (GS). The second generation of cells were divided into control group, high glucose group, oxidative stress group and hypoxia group randomly. The morphological changes of the cells were observed under inverted microscope, the changes of GS and α-smooth muscle actin (α-SMA) were observed by immunofluorescence staining, and the cell migration ability was observed by Transwell chamber. Results The Müller cells were successfully cultured and identified in vitro. In all groups, the function of Müller cells was impaired, and in oxidative stress group and hypoxia group, the structure was damaged, and the normal morphology was lost. In hypoxia group, the expression of α-SMA was positive, and the number of cells in the Transwell chamber was significantly higher than that in other groups, the differences were statistically significant (P < 0.05). Conclusion High glucose, oxidative stress and hypoxia can destroy the function of Müller cells. Hypoxia can induce transforming into myofibroblast cells with migratory ability, which indicates that Müller cells are involved in the pathogenesis of proliferative diabetic retinopathy.