Thioredoxin 1 overexpression attenuated diabetes-induced endoplasmic reticulum stress in Müller cells via apoptosis signal-regulating kinase 1.

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.

Upregulation of thioredoxin contributes to inhibiting diabetic hearing impairment.

AIMS: Hair cell reduction was related to diabetes-induced hearing loss. Oxidative stress, endoplasmic reticulum stress, and autophagy participate in this process. Thioredoxin (Trx) is a protein with many biological functions which can regulate them. In this study, aiming to clarify protective effect of Trx on diabetic hearing loss and to identify an early potential therapeutic target for diabetic hearing impairment in the future. METHODS: Trx transgenic (Tg) mice were used to establish a diabetic model by intraperitoneally injecting streptozotocin (STZ) and with/without SF or PX12 treatment. Succinate dehydrogenase (SDH) staining was used to evaluate the loss of hair cells. The relative expression of related proteins and genes was detected using western blotting and qRT-PCR. RESULTS: In vivo, loss of outer hair cells was observed. However, it can be delayed Trx overexpression. Moreover, the expression of PGC-1α, bcl-2 and LC3 was increased in Tg(+)-DM mice compared with Tg(-)-DM mice. The expression of ASK1, Txnip, GRP78, CHOP and p62 was decreased in Tg(+)-DM mice compared with Tg(-)-DM mice. CONCLUSIONS: Upregulation of Trx protects diabetes-induced cochlear hair cells reduction. The underlying mechanisms were related to the regulation of ER stress through ASK1 and the mitochondrial pathway or autophagy via Txnip.

Stimulation of AMPK Prevents Diabetes-Induced Photoreceptor Cell Degeneration.

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.

Ginkgo biloba delays light-induced photoreceptor degeneration through antioxidant and antiapoptotic properties.

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.

Sulforaphane delays diabetes-induced retinal photoreceptor cell degeneration.

Diabetic retinopathy (DR) is a serious neurodegenerative disease that is induced by hyperglycaemia. Oxidative stress, inflammation and endoplasmic reticulum (ER) stress are involved in the development of DR. Sulforaphane (SF) is widely found in cruciferous plants and has a protective effect against retinal neurodegeneration in diabetes, but the mechanism is unclear. In this study, we investigated the mechanism by which SF protects against photoreceptor degeneration in diabetes. In vivo, a mouse model of diabetes was established by streptozotocin (STZ) injection, and the mice were treated with/without SF. Electroretinography (ERG) and H&E staining were used to evaluate retinal function and morphology. In vitro, 661w cells were treated with AGEs with/without SF. Cell viability and apoptosis were analysed by CCK-8 assay and flow cytometry. The expression of proteins and genes was assessed by western blot and qRT-PCR. The amplitude of the a-wave was decreased and the morphology was changed in the diabetic mice, and these changes were delayed by SF treatment. The percentage of apoptotic cells was increased and the cell viability was decreased after the treatment of 661w cells with AGEs. Moreover, the expression of GRP78, Txnip and TNFα was increased, however, this increased expression was reversed by SF treatment via AMPK pathway activation. Taken together, these data show that SF can delay photoreceptor degeneration in diabetes, and the underlying mechanism is related to the inhibition of ER stress, inflammation and Txnip expression through the activation of the AMPK pathway.