Ophthalmology Unit, Department of Health Sciences, Azienda Ospedaliera Universitaria Maggiore della Carità, University of East Piedmont, Novara, Italy.
Lab. Physiology/Experimental Surgery, Department of Translational Medicine, University of East Piedmont, Novara, Italy.
Acta Ophthalmol. 2019 Jun;97(4):e559-e567. doi: 10.1111/aos.13995. Epub 2018 Dec 26.
Subthreshold micropulse laser (SMPL) has been increasingly used for the treatment of different retinal and choroidal macular disorders. However, the exact mechanisms of action have not yet been clearly defined. Therefore, we aimed to examine the role of SMPL treatment in the modulation of oxidant/antioxidant systems, apoptosis and autophagy in the mice eyes.
A specific laser contact lens for retina was positioned on the cornea of 40 mice (20 young and 20 old) in order to focus the laser on the eye fundus for SMPL treatment. Within 6 months, 20 animals received one treatment only, whereas the others were treated three times. Eye specimens underwent histological analysis and were used for thiobarbituric acid reactive substances (TBARS) and glutathione (GSH) quantification, as well as for the superoxide dismutase 1 (SOD1) and the selenoprotein thioredoxin reductase 1 (TrxR1) expression evaluation. Western blot was performed for nitric oxide synthase (NOS) subtypes detection and to examine changes in apoptotic/autophagy proteins expression.
SMPL treatment reduced TBARS and increased GSH and SOD1 in the mice eyes. It also reduced cytochrome c, caspase 3 expression and activity and cleaved caspase 9, and increased Beclin 1, p62 and LC3β. The effects were more relevant in the elderly animals.
Our results showed that SMPL therapy restored the oxidant/antioxidant balance within retinal layers and modulated programmed forms of cell death. Further studies may confirm these data and could evaluate their relevance in clinical practice.
阈下微脉冲激光(SMPL)已越来越多地用于治疗不同的视网膜和脉络膜黄斑疾病。然而,其确切的作用机制尚未明确。因此,我们旨在研究 SMPL 治疗在调节氧化应激/抗氧化系统、凋亡和自噬方面在小鼠眼中的作用。
将特定的用于视网膜的激光接触镜放置在 40 只小鼠(20 只年轻和 20 只年老)的角膜上,以便将激光聚焦在眼底进行 SMPL 治疗。在 6 个月内,20 只动物仅接受一次治疗,而其余动物则接受三次治疗。对眼部标本进行组织学分析,并用于测定硫代巴比妥酸反应物质(TBARS)和谷胱甘肽(GSH)的含量,以及超氧化物歧化酶 1(SOD1)和硒蛋白硫氧还蛋白还原酶 1(TrxR1)的表达评估。通过 Western blot 检测一氧化氮合酶(NOS)亚型的表达,并检测凋亡/自噬蛋白表达的变化。
SMPL 治疗降低了小鼠眼内的 TBARS 并增加了 GSH 和 SOD1。它还降低了细胞色素 c、半胱天冬酶 3 的表达和活性以及切割的半胱天冬酶 9,并增加了 Beclin 1、p62 和 LC3β。这些作用在老年动物中更为明显。
我们的结果表明,SMPL 治疗恢复了视网膜各层的氧化应激/抗氧化平衡,并调节了程序性细胞死亡。进一步的研究可能会证实这些数据,并评估其在临床实践中的相关性。