No abstract

Diabetic retinopathy (DR) is a kind of severe retinal neurodegeneration. The advanced glycation end products (AGEs) affect autophagy, and mitochondrial function is involved in DR. Adenosine-activated protein kinase (AMPK) is an important metabolic sensor that can regulate energy homeostasis in cells. However, the effect of AMPK in DR is still not fully understood. In this study, we investigated the effect of AMPK on diabetes-induced photoreceptor cell degeneration. In vivo, a diabetic mouse model was established by streptozotocin (STZ) injection. Haematoxylin-eosin (HE) staining was used to observe retinal morphology and measure the thicknesses of different layers in the retina. Electroretinogram (ERG) was used to evaluate retinal function. In vitro, 661w cells were treated with AGEs with/without an AMPK agonist (metformin) or AMPK inhibitor (compound C). Flow cytometry and CCK-8 assays were used to analyse apoptosis. Mitochondrial membrane potential was analysed by JC-1. Western blotting and qRT-PCR were used to examine the expression of related proteins and genes, respectively. The wave amplitude and the thickness of the outer nuclear layer were decreased in diabetic mice. The expression of rhodopsin and opsin was also decreased in diabetic mice. In vitro, the percentage of apoptotic cells was increased, the expression of the apoptosis-related protein Bax was increased, and Bcl-2 was decreased after AGE treatment in 661w cells. The expression of the autophagy-related protein LC3 was decreased, and p62 was increased. The mitochondrial-related gene expression and membrane potential were decreased, and mitochondrial morphology was abnormal, as observed by TEM. However, AMPK stimulation ameliorated this effect. These results indicate that AMPK stimulation can delay diabetes-induced photoreceptor degeneration by regulating autophagy and mitochondrial function.

Intense exposure to artificial bright light increases the risk of retinal damage resulting in blurred vision and blindness. Long-term exposure to bright light elevates oxidative stress-induced apoptosis, which results in photoreceptor cell degeneration. However, to the best of our knowledge, the molecular mechanism associated with light-induced retinopathy remains unclear. In the present study, the mechanisms involved in light-induced oxidative stress and apoptosis were investigated along with the protective effects of Ginkgo biloba (EGb 761) in photoreceptor cell degeneration. EGb 761 was administered to mice at a dose of 50 or 100 mg/kg for 7 days prior to exposure to bright light (5,000 lux for 24 h). Furthermore, photoreceptor cell disorders were evaluated using electroretinogram (ERG) and H&E staining analyses. The expression levels of antioxidant genes and proteins ERK, thioredoxin (Trx) and nuclear factor erythroid 2-related factor 2 (Nrf-2) and the induction of apoptosis cytochrome c (Cyc), cleaved caspase-3 and Bax, were determined by reverse transcription-quantitative PCR and western blotting. ERG and histological analysis revealed that exposure to bright light induced functional and morphological changes to the photoreceptor cells. Exposure to bright light increased the levels of Cyc, cleaved caspase-3 and Bax, and decreased the levels of phosphorylated (p-) Erk, Nrf-2 and thioredoxin (Trx). However, treatment of mice with EGb 761 increased the expression levels of antiapoptotic (Bcl-2) and antioxidant (p-Erk, Trx and Nrf-2) proteins and decreased the expression levels of the apoptotic genes (Cyc, cleaved caspase-3 and Bax). Based on these findings, the present study suggested that prolonged exposure to light induces photoreceptor cell degeneration, where EGb 761 treatment may serve a therapeutic effect on the development of photoreceptor cell degeneration.

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As one of the common and serious chronic complications of diabetes mellitus (DM), the related mechanism of diabetic retinopathy (DR) has not been fully understood. Müller cell reactive gliosis is one of the early pathophysiological features of DR. Therefore, exploring the manner to reduce diabetes‐induced Müller cell damage is essential to delay DR. Thioredoxin 1 (Trx1), one of the ubiquitous redox enzymes, plays a vital role in redox homeostasis via protein–protein interactions, including apoptosis signal‐regulating kinase 1 (ASK1). Previous studies have shown that upregulation of Trx by some drugs can attenuate endoplasmic reticulum stress (ERS) in DR, but the related mechanism was unclear. In this study, we used DM mouse and high glucose (HG)‐cultured human Müller cells as models to clarify the effect of Trx1 on ERS and the underlying mechanism. The data showed that the diabetes‐induced Müller cell damage was increased significantly. Moreover, the expression of ERS and reactive gliosis was also upregulated in diabetes in vivo and in vitro. However, it was reversed after Trx1 overexpression. Besides, ERS‐related protein expression, reactive gliosis, and apoptosis were decreased after transfection with ASK1 small‐interfering RNA in stable Trx1 overexpression Müller cells after HG treatment. Taken together, Trx1 could protect Müller cells from diabetes‐induced damage, and the underlying mechanism was related to inhibited ERS via ASK1.