Effects of an Aquaporin 4 Inhibitor, TGN-020, on Murine Diabetic Retina
Effects of an Aquaporin 4 Inhibitor, TGN-020, on Murine Diabetic Retina
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DOI：
10.3390/ijms21072324
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发表时间：
2020-04-01
影响因子：
5.6
通讯作者：
Ikeda, Tsunehiko
中科院分区：
文献类型：
--
作者：
Oosuka, Shou;Kida, Teruyo;Ikeda, Tsunehiko
目的：目的探讨水通道蛋白4（AQP 4）抑制剂2-烟酰胺-1，3，4-噻二唑（TGN-020）对糖尿病视网膜血管内皮生长因子（VEGF）表达、活性氧（ROS）产生及视网膜水肿的影响。方法：对链脲佐菌素诱导的糖尿病大鼠玻璃体内注射贝伐单抗、TGN-020或磷酸盐缓冲液（PBS）。对视网膜切片进行抗胶质细胞酸性蛋白（GFAP）、抗AQP 4和抗VEGF免疫染色。通过Western印迹分析测定来自收集的视网膜的VEGF蛋白水平。此外，在存在或不存在TGN-020的情况下，在糖尿病大鼠的平板视网膜中观察到伊文思蓝的视网膜血管渗漏。在生理和高葡萄糖条件下，在存在或不存在TGN-020或贝伐珠单抗的情况下，使用乙锭荧光的流式细胞术分析测定大鼠视网膜Muller细胞（TR-MUL 5;转基因大鼠Muller细胞）的体积变化和细胞内ROS水平。结果：TGN-020可抑制糖尿病视网膜VEGF的免疫反应性和蛋白水平。AQP 4免疫反应性高于对照组，且与GFAP共定位。与VEGF类似，AQP 4和GFAP也被TGN-020抑制。在伊文思蓝测定中，TGN-020减少了糖尿病视网膜中的渗漏。在培养的Muller细胞中，暴露于TGN-020与暴露于贝伐珠单抗一样抑制了高糖条件下细胞体积和细胞内ROS产生的增加。结论：TGN-020对糖尿病视网膜水肿有抑制作用。
Purpose: To investigate the effect of a selective aquaporin 4 (AQP4) inhibitor, 2-(nicotinamide)-1,3,4-thiadiazole (TGN-020), on the expression of vascular endothelial growth factor (VEGF) and reactive oxygen species (ROS) production, as well as on the retinal edema in diabetic retina. Methods: Intravitreal injections of bevacizumab, TGN-020, or phosphate-buffered saline (PBS) were performed on streptozotocin-induced diabetic rats. Retinal sections were immunostained for anti-glial fibrillary acidic protein (GFAP), anti-AQP4, and anti-VEGF. Protein levels of VEGF from collected retinas were determined by Western blot analysis. In addition, retinal vascular leakage of Evans Blue was observed in the flat-mounted retina from the diabetic rats in the presence or absence of TGN-020. Volumetric changes of rat retinal Muller cells (TR-MUL5; transgenic rat Muller cells) and intracellular levels of ROS were determined using flow cytometry analysis of ethidium fluorescence in the presence or absence of TGN-020 or bevacizumab under physiological and high glucose conditions. Results: In the diabetic retina, the immunoreactivity and protein levels of VEGF were suppressed by TGN-020. AQP4 immunoreactivity was higher than in the control retinas and the expressions of AQP4 were co-localized with GFAP. Similarly to VEGF, AQP4 and GFAP were also suppressed by TGN-020. In the Evans Blue assay, TGN-020 decreased leakage in the diabetic retinas. In the cultured Muller cells, the increase in cell volumes and intracellular ROS production under high glucose condition were suppressed by exposure to TGN-020 as much as by exposure to bevacizumab. Conclusion: TGN-020 may have an inhibitory effect on diabetic retinal edema.