Oxygen glucose deprivation/re-oxygenation-induced neuronal cell death is associated with Lnc-D63785 m6A methylation and miR-422a accumulation

Abstract

Oxygen glucose deprivation/re-oxygenation (OGD/R) induces neuronal injury via mechanisms that are believed to mimic the pathways associated with brain ischemia. In SH-SY5Y cells and primary murine neurons, we report that OGD/R induces the accumulation of the microRNA miR-422a, leading to downregulation of miR-422a targets myocyte enhancer factor-2D (MEF2D) and mitogen-activated protein kinase kinase 6 (MAPKK6). Ectopic miR-422a inhibition attenuated OGD/R-induced cell death and apoptosis, whereas overexpression of miR-422a induced significant neuronal cell apoptosis. In addition, OGD/R decreased the expression of the long non-coding RNA D63785 (Lnc-D63785) to regulate miR-422a accumulation. Lnc-D63785 directly associated with miR-422a and overexpression of Lnc-D63785 reversed OGD/R-induced miR-422a accumulation and neuronal cell death. OGD/R downregulated Lnc-D63785 expression through increased methyltransferase-like protein 3 (METTL3)-dependent Lnc-D63785 m6A methylation. Conversely METTL3 shRNA reversed OGD/R-induced Lnc-D63785 m6A methylation to decrease miR-422a accumulation. Together, Lnc-D63785 m6A methylation by OGD/R causes miR-422a accumulation and neuronal cell apoptosis.

In Silico Identification and Mechanism Exploration of Active Ingredients against Stroke from An-Gong-Niu-Huang-Wan (AGNHW) Formula

“…By comprehensively considering the network-based predicted ranking, chemical structure, physicochemical properties, availability, and accessibility, these promising ingredients were selected for antistroke efficacy evaluation ( Figure 5 ). Subsequently, the OGD/R injury model, a recognized in vitro model in ischemic stroke research, was applied for pharmacological validation [ 46 , 47 ]. As shown in Figure 6 , the OGD/R insult group showed significantly lowered PC12 cell viability ( P < 0.01), while CA (6.25, 12.5, and 25 μ M), SCU (1, 2, and 4 μ M), chrisin (1, 2, and 4 μ M), fisetin (12 μ M), gallic acid (6.25 μ M), imperatorin (1, 2, and 4 μ M), and palmatine (3.125, 6.25, and 12.5 μ M) increased the viability of PC12 cells ( ∗ P < 0.05, ∗∗ P < 0.01, and ∗∗∗ P < 0.001, respectively).…”

Transcriptome-Wide High-Throughput m6A Sequencing of Differential m6A Methylation Patterns in the Human Rheumatoid Arthritis Fibroblast-Like Synoviocytes Cell Line MH7A

“…5 , 7 Various m6A demethylases are called “erasers”, including fat mass and obesity-associated protein (FTO) and ALKB family member 5 (ALKBH5), which function is to reduce the modified RNA to the original RNA. 8 In addition, m6A binding protein acts as a “reader” and mainly includes the YT521-B homology (YTH) and heterogeneous nuclear ribonucleoprotein (HNRNP) families, such as HNRNPC, YTHDC1, YTHDC2, YTHDF1 and YTHDF2, which recognize m6A-modified RNA and thus regulate mRNA metabolism and function. 9 , 10 …”

m6A Modification in Mammalian Nervous System Development, Functions, Disorders, and Injuries

“…The expression of m 6 A erasers, ALKBH5 and FTO, are decreased but not writers. Overexpression of m 6 A erasers can alleviate neuronal damage induced by I/R injury ( Xu S. et al, 2020 ). A third study found that oxygen-glucose deprivation/re-oxygenation (OGD/R) increased METTL3-dependent m 6 A modification of long non-coding RNA D63785 (lnc-D63785), thus causing reduced expression of lnc-D63785 ( Xu S. et al, 2020 ).…”

“…Overexpression of m 6 A erasers can alleviate neuronal damage induced by I/R injury ( Xu S. et al, 2020 ). A third study found that oxygen-glucose deprivation/re-oxygenation (OGD/R) increased METTL3-dependent m 6 A modification of long non-coding RNA D63785 (lnc-D63785), thus causing reduced expression of lnc-D63785 ( Xu S. et al, 2020 ). Downregulation of lnc-D63785 further induces accumulation of miR-422a, which results in neuronal cell apoptosis ( Xu S. et al, 2020 ).…”