Proliferation and migration of retinal endothelial cells (RECs) contribute to the development of diabetic retinopathy. PLAG1 (pleomorphic adenoma gene 1) functions as a zinc-finger transcription factor to participate in the development of lipoblastomas or pleomorphic adenomas of the salivary glands through regulation of cell proliferation and migration. The role of PLAG1 in diabetic retinopathy was investigated in this study. Firstly, RECs were induced under high glucose conditions, which caused reduction in viability and induction of apoptosis in the RECs. Indeed, PLAG1 was elevated in high glucosetreated RECs. Functional assays showed that silence of PLAG1 increased viability and suppressed apoptosis in high glucose-induced RECs, accompanied with up-regulation of Bcl-2 and down-regulation of Bax and cleaved caspase-3. Moreover, migration of RECs was promoted by high glucose conditions, while repressed by knockdown of PLAG1. High glucose also triggered angiogenesis of RECs through up-regulation of vascular endothelial growth factor (VEGF). However, interference of PLAG1 reduced VEGF expression to retard the angiogenesis. Silence of PLAG1 also attenuated high glucose-induced up-regulation of Wnt3a, β-catenin and c-Myc in RECs. Moreover, silence of PLAG1 ameliorated histopathological changes in the retina of STZ-induced diabetic rats through down-regulation of β-catenin. In conclusion, knockdown of PLAG1 suppressed high glucose-induced angiogenesis and migration of RECs, and attenuated diabetic retinopathy by inactivation of Wnt/ β-catenin signalling.
- MeSH
- beta-katenin metabolismus MeSH
- diabetická retinopatie * MeSH
- DNA vazebné proteiny metabolismus MeSH
- endoteliální buňky metabolismus MeSH
- experimentální diabetes mellitus * MeSH
- glukosa toxicita MeSH
- kaspasa 3 MeSH
- krysa rodu rattus MeSH
- patologická angiogeneze MeSH
- protein X asociovaný s bcl-2 MeSH
- retina metabolismus MeSH
- signální dráha Wnt MeSH
- transkripční faktory MeSH
- vaskulární endoteliální růstový faktor A MeSH
- zinek MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Diabetic or hyperglycaemic conditions stimulate the inflammatory response, excessive accumulation of extracellular matrix, and result in glomerulosclerosis, a scarring process of diabetic nephropathy. c-Jun activation domain-binding protein 1 (JAB1) functions as a regulator of pathways involved in cellular apoptosis and proliferation. The role of JAB1 in diabetic nephropathy was investigated in this study. Firstly, glomerular mesangial cells (GMCs) were treated with high glucose, and high glucose conditions induced up-regulation of JAB1 in the GMCs. Moreover, IL-6, TNF-α, MCP-1, and IL-1β were also elevated in high glucose-induced GMCs. Secondly, silencing of JAB1 reduced the levels of IL-6, TNF-α, MCP-1, and IL-1β in high glucose-induced GMCs. In addition, silencing of JAB1 attenuated the high glucose-induced decrease of superoxide dismutase (SOD) and the increase of reactive oxygen species (ROS) and malondialdehyde (MDA). The increased TGF-β1, collagen I, collagen IV, and fibronectin levels in high glucose-induced GMCs were restored by knockdown of JAB1. Thirdly, angiopoietin-like protein 2 (ANGPTL2) expression was reduced by JAB1. Over-expression of ANGPTL2 weakened the JAB1 silence-induced decrease of IL-6, TNF-α, MCP-1, IL-1β, TGF-β1, collagen I, collagen IV, and fibronectin. In conclusion, silencing of JAB1 reduced extracellular matrix deposition and suppressed inflammation in high glucose-induced GMCs through down-regulation of ANGPTL2.
- MeSH
- angiopoetinu podobné proteiny metabolismus MeSH
- angiopoetinu podobný protein 2 MeSH
- diabetické nefropatie * metabolismus MeSH
- extracelulární matrix metabolismus MeSH
- fibronektiny metabolismus MeSH
- glukosa metabolismus toxicita MeSH
- interleukin-6 MeSH
- kolagen metabolismus MeSH
- lidé MeSH
- mesangiální buňky * metabolismus MeSH
- signální transdukce MeSH
- TNF-alfa metabolismus MeSH
- transformující růstový faktor beta1 metabolismus MeSH
- zánět MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
Diabetes increases the risk and worsens the progression of cognitive impairment. The hippocampus is an important domain for learning and memory. We previously showed that endothelin-1 (ET-1) reduced diabetes-induced inflammation in hippocampal neurons, suggesting a neuroprotective effect. Given that neurons and endothelial cells within the neurovascular unit depend on each other for proper function, we investigated the effect of ET-1 on brain-derived neurotrophic factor (BDNF) synthesis, a key neurotrophin and prosurvival factor, in neuronal (HT22 hippocampal neurons) and brain microvascular endothelial (BMEC-5i) cells under normal and diabetes-mimicking (high glucose plus palmitate) conditions. Cells were treated with exogenous ET-1 or ET receptor antagonists including ET(B) receptor selective antagonist BQ788 (1 microM) or dual-receptor antagonist bosentan (10 microM). Mature (m)BDNF, proBDNF and caspase-3 levels were measured by Western blotting. Diabetic conditions reduced the prosurvival mBDNF/proBDNF ratio in both HT22 and BMEC-5i cells. Addition of exogenous ET-1 had no effect on the BDNF system in HT22 cells in diabetic conditions. Both HT22 and BMEC-5i cells had an increase in the mBDNF/proBDNF ratio when grown in diabetes-simulating conditions in the presence of endothelin receptor inhibition. These data suggest that blockade of ET-1 may provide neuroprotection to hippocampal cells through the modulation of the BDNF system.
- MeSH
- endoteliální buňky účinky léků metabolismus MeSH
- endotelin-1 antagonisté a inhibitory metabolismus MeSH
- glukosa toxicita MeSH
- hipokampus cytologie účinky léků metabolismus MeSH
- mozkový neurotrofický faktor metabolismus MeSH
- myši MeSH
- neurony účinky léků metabolismus MeSH
- palmitany toxicita MeSH
- transformované buněčné linie MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Endothelial cells (ECs) are primary targets of glucose-induced tissue damage. As a result of hyperglycemia, endothelin-1 (ET-1) is upregulated in organs affected by chronic diabetic complications. The objective of the present study was to identify novel transcriptional mechanisms that influence ET-1 regulation in diabetes. We carried out the investigation in microvascular ECs using multiple approaches. ECs were incubated with 5 mM glucose (NG) or 25 mM glucose (HG) and analyses for DNA methylation, histone methylation, or long non-coding RNA- mediated regulation of ET-1 mRNA were then performed. DNA methylation array analyses demonstrated the presence of hypomethylation in the proximal promoter and 5' UTR/first exon regions of EDN1 following HG culture. Further, globally blocking DNA methylation or histone methylation significantly increased ET-1 mRNA expressions in both NG and HG-treated HRECs. While, knocking down the pathogenetic lncRNAs ANRIL, MALAT1, and ZFAS1 subsequently prevented the glucose-induced upregulation of ET-1 transcripts. Based on our past and present findings, we present a novel paradigm that reveals a complex web of epigenetic mechanisms regulating glucose-induced transcription of ET-1. Improving our understanding of such processes may lead to better targeted therapies.
- MeSH
- endoteliální buňky účinky léků metabolismus MeSH
- endotelin-1 agonisté fyziologie MeSH
- epigeneze genetická účinky léků fyziologie MeSH
- experimentální diabetes mellitus chemicky indukované metabolismus MeSH
- glukosa toxicita MeSH
- komplikace diabetu genetika metabolismus MeSH
- kultivované buňky MeSH
- lidé MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- MeSH
- biologický transport MeSH
- diabetes mellitus 2. typu patofyziologie prevence a kontrola MeSH
- glukosa krev metabolismus toxicita MeSH
- hyperglykemie etiologie MeSH
- inzulin krev sekrece MeSH
- lidé MeSH
- lipidy krev metabolismus toxicita MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- srovnávací studie MeSH
- Geografické názvy
- Spojené státy americké MeSH
- MeSH
- biologický transport MeSH
- diabetes mellitus 2. typu dietoterapie etiologie farmakoterapie MeSH
- glukosa krev metabolismus toxicita MeSH
- hyperglykemie MeSH
- inzulin krev sekrece MeSH
- lidé MeSH
- lipidy krev metabolismus toxicita MeSH
- Check Tag
- lidé MeSH
- Geografické názvy
- Spojené státy americké MeSH
