UNASSIGNED: Hematoxylin and eosin staining were used to observe the morphology of the retina. Immunofluorescence was used to detect positive (Nrf2+ and VEGF+) cells. Immunohistochemistry was used to detect the level of nuclear expression of PCNA in retinal tissue. Transmission electron microscope (TEM) was used to observe the morphology and structure of pigment cells. qRT-PCR was used to assay the expression of miR-23a-3p, Nrf2, and HO-1. Western blotting was used to detect the expression of Nrf2, HO-1, β-actin, and Lamin B1.
UNASSIGNED: Melatonin or miR-23a-3p antagomir treatment could ameliorate the Oxygen-induced pathological changes, increased the expression of Nrf2 and HO-1, SOD, and GSH-Px, and decreased the expression of VEGF, miR-23a-3p, MDA and the apoptosis in the ROP model. Further target prediction and luciferase reporter assays confirmed the targeted binding relationship between miR-23a-3p and Nrf2.
UNASSIGNED: Our study showed that melatonin could ameliorate H2O2-induced apoptosis and oxidative stress injury in RGC cells by mediating miR-23a-3p/Nrf2 signaling pathway, thereby improving retinal degeneration.
■苏木精和伊红染色用于观察视网膜的形态。免疫荧光法检测阳性(Nrf2+和VEGF+)细胞。免疫组化法检测视网膜组织中PCNA的核表达水平。透射电镜(TEM)观察色素细胞的形态和结构。qRT-PCR用于检测miR-23a-3p的表达,Nrf2和HO-1。蛋白质印迹法检测Nrf2、HO-1、β-肌动蛋白的表达,还有LaminB1.
■褪黑素或miR-23a-3pantagomir治疗可以改善氧诱导的病理变化,Nrf2和HO-1,SOD的表达增加,和GSH-Px,并降低VEGF的表达,miR-23a-3p,ROP模型中MDA和细胞凋亡。进一步的靶标预测和荧光素酶报告基因测定证实了miR-23a-3p和Nrf2之间的靶向结合关系。
■我们的研究表明,褪黑素可以通过介导miR-23a-3p/Nrf2信号通路改善H2O2诱导的RGC细胞凋亡和氧化应激损伤,从而改善视网膜变性。