Aldose reductase inhibitor fidarestat counteracts diabetes-associated cataract formation, retinal oxidative-nitrosative stress, glial activation, and apoptosis.
Aldose reductase inhibitor fidarestat counteracts diabetes-associated cataract formation, retinal oxidative-nitrosative stress, glial activation, and apoptosis.
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DOI：
10.3892/ijmm.21.6.667
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发表时间：
2008-06
影响因子：
5.4
通讯作者：
V. Drel;P. Pacher;T. K. Ali;Jeho Shin;U. Julius;A. El-Remessy;I. Obrosova
中科院分区：
文献类型：
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作者：
V. Drel;P. Pacher;T. K. Ali;Jeho Shin;U. Julius;A. El-Remessy;I. Obrosova
这项研究旨在评估与糖尿病相关的白内障形成以及视网膜氧化硝化应激，神经胶质激活和细胞凋亡的有效和特异性醛糖还原酶抑制剂Fidarestat。对照和链霉菌素糖尿病大鼠接受或没有Fidarestat（16 mg kg（-1）d（-1））的治疗，持续2周后10周，未经治疗。通过间接眼镜检查和便携式缝隙灯评估镜头变化。通过免疫组织化学评估硝基酪氨酸，聚（ADP-核糖）和神经胶质纤维酸性蛋白表达。通过免疫过氧化物酶染色，通过TUNEL分析在扁平的视网膜中定量凋亡率。为了剖析视网膜微血管细胞中高葡萄糖暴露的作用，将原发性牛视网膜周周细胞和内皮细胞在5或30 mM葡萄糖中培养，有或没有Fidarestat（10 microm）3-14天。通过TUNEL分析，硝基酪氨酸和聚（ADP-核糖）通过免疫细胞化学评估凋亡，并通过蛋白质印迹分析评估BAX和BCL-2表达。 Fidarestat治疗可防止糖尿病性白内障形成和抵消视网膜亚硝化应激，以及聚（ADP-核糖）聚合酶激活以及神经胶质激活。在糖尿病大鼠与对照组（207 +/- 33 vs. 49 +/- 4，p <0.01）中，TUNEL阳性核（平均+/- SEM）的数量增加了约4倍，并且这种增加是部分增加的Fidarestat（106 +/- 34，p <0.05 vs.未经治疗的糖尿病组）预防。凋亡细胞数量随着周细胞和内皮细胞暴露至高葡萄糖水平的延长而增加。 Fidarestat在两种细胞类型中应对亚硝基酪氨酸和聚（ADP-核糖）积累和凋亡。 Fidarestat在高葡萄糖暴露于视网膜周细胞中的抗凋亡作用与BAX抑制或Bcl-2表达的增加无关。总之，研究结果i）支持醛糖还原酶在糖尿病相关的白内障形成中的重要作用，以及视网膜氧化硝化应激，神经胶质激活和凋亡以及ii）为发展醛糖糖还原酶抑制剂的发展提供了理由。特别是，用于预防和治疗糖尿病眼并发症。
This study was aimed at evaluating the potent and specific aldose reductase inhibitor fidarestat, on diabetes-associated cataract formation, and retinal oxidative-nitrosative stress, glial activation, and apoptosis. Control and streptozotocin-diabetic rats were treated with or without fidarestat (16 mg kg(-1)d(-1)) for 10 weeks after an initial 2-week period without treatment. Lens changes were evaluated by indirect ophthalmoscopy and portable slit lamp. Nitrotyrosine, poly(ADP-ribose), and glial fibrillary acidic protein expression were assessed by immunohistochemistry. The rate of apoptosis was quantified in flat-mounted retinas by TUNEL assay with immunoperoxidase staining. To dissect the effects of high glucose exposure in retinal microvascular cells, primary bovine retinal pericytes and endothelial cells were cultured in 5 or 30 mM glucose, with or without fidarestat (10 microM) for 3-14 days. Apoptosis was assessed by TUNEL assay, nitrotyrosine and poly(ADP-ribose) by immunocytochemistry, and Bax and Bcl-2 expression by Western blot analyses. Fidarestat treatment prevented diabetic cataract formation and counteracted retinal nitrosative stress, and poly(ADP-ribose) polymerase activation, as well as glial activation. The number of TUNEL-positive nuclei (mean +/- SEM) was increased approximately 4-fold in diabetic rats vs. controls (207+/-33 vs. 49+/-4, p<0.01), and this increase was partially prevented by fidarestat (106+/-34, p<0.05 vs. untreated diabetic group). The apoptotic cell number increased with the prolongation of exposure of both pericytes and endothelial cells to high glucose levels. Fidarestat counteracted nitrotyrosine and poly(ADP-ribose) accumulation and apoptosis in both cell types. Antiapoptotic effect of fidarestat in high glucose-exposed retinal pericytes was not associated with the inhibition of Bax or increase in Bcl-2 expression. In conclusion, the findings, i) support an important role for aldose reductase in diabetes-associated cataract formation, and retinal oxidative-nitrosative stress, glial activation, and apoptosis, and ii) provide a rationale for the development of aldose reductase inhibitors, and, in particular, fidarestat, for the prevention and treatment of diabetic ocular complications.