Project description:Brain microvascular endothelial cell (BMEC) injury can affect neuronal survival by modulating immune responses through the microenvironment. We used microarrays to detail the miRNAs expression in the exosomes from normal and oxygen glucose deprivation (OGD)-cultured BMECs.
Project description:The present study has used whole-rat genome microarray expression profiling to identify genes whose expression is significantly altered in hippocampal neuronal cultures submitted to oxygen and glucose deprivation (OGD), an established in vitro model for cerebral global ischemia that is suitable for investigations at the molecular level. To do so, total RNA was extracted from hippocampal neuronal cultures at an early (7h) and delayed (24h) time point after OGD, as well as from control neurons. Analysis of gene ontology showed that OGD followed by 7h or 24h of recovery induces changes in the expression levels of genes related with inflammation, response to oxidative stress, metabolism, apoptosis, synaptic proteins and ion channels and, importantly, genes that show different expression levels are mainly specific to one of the two time points of recovery analyzed. The expression levels of several genes were confirmed by qPCR and were in good agreement with the microarray data, showing that the combined use of the OGD model and the microarray technology can be a useful tool for the study molecular mechanisms contributing to the neuronal demise after transient global ischemia. Ischemia induced gene expression in primary hippocampal neuronal rat cultures was measured at 7 and 24 hours after exposure to Oxygen and Glucose deprivation (OGD). Three independent experiments were performed at each time (7 or 24 hours) as independent biological replicates.
Project description:Oxygen and glucose metabolism plays a pivotal role in many (patho)physiological conditions. In particular, oxygen and glucose deprivation (OGD) occurs during ischemia and stroke, resulting in extensive tissue injury and cell death. We applied time-resolved ribosome profiling technique to assess early events at the level of gene expression in rat pheochromocytoma PC12 cells during short-term OGD. Most substantial alterations in transcripts levels and their translation were seen to occur in the first 20 minutes of OGD. The rapid adaptation of translation apparatus to OGD is global and involves altered elongation and initiation rates. We also observed salient and reproducible alterations in ribosome densities of individual mRNAs such as increased translation of particular upstream Open Reading Frames (uORFs); induced site-specific arrests of the ribosomes and synthesis of extended protein isoforms. Ribosome profiling (with mRNA-seq sequencing) was carried out at 0,20,40 and 60 minutes of OGD. Two biological replicates were used.
Project description:Ischemia‑reperfusion (I/R) injury is a key pathological process in myocardial infarction, for which effective therapies are still lacking. Phillyrin has been reported to exert cardioprotective effects, but its molecular mechanisms remain unclear. HL‑1 mouse cardiomyocytes were subjected to oxygen‑glucose deprivation/reoxygenation (OGD/R) to mimic I/R injury. Cells were divided into two groups: OGD/R alone (IR) and OGD/R with Phillyrin treatment (IR+Phil). Total RNA was extracted from 3 biological replicates per group and sequenced using Illumina NovaSeq 6000. Transcriptomic profiling identified differentially expressed genes and pathways regulated by OGD/R and Phillyrin treatment. The processed data file contains raw read counts and FPKM values for all genes across all samples. This dataset provides a comprehensive transcriptomic resource to explore the molecular mechanisms of Phillyrin‑mediated cardioprotection against I/R injury.
Project description:Background: Cerebral infarction leads to blood-brain barrier (BBB) disruption, exacerbating brain injury through edema, inflammation, and neuronal death. Although BBB damage is a critical event in stroke pathology, the underlying molecular mechanisms and reliable biomarkers remain poorly understood. This study aimed to identify key biomarkers associated with BBB injury following cerebral infarction using an in vitro model and transcriptomic approaches. Human cerebral microvascular endothelial cells (hCMEC/D3) were subjected to oxygen-glucose deprivation (OGD) and OGD/reoxygenation (OGD/R) to simulate ischemic and reperfusion injury. Cell viability, inflammatory cytokines, LDH release, and angiogenesis were assessed. Transcriptomic sequencing, weighted gene co-expression network analysis (WGCNA), and random forest algorithms were employed to identify differentially expressed genes and key biomarkers. OGD treatment significantly increased IL-1β, IL-6, TNF-α, and LDH levels, which were partially reversed by OGD/R. Transcriptomic analysis identified 1229 and 800 differentially expressed genes respectively in OGD and OGD/R comparisons. Enrichment analysis highlighted ribosome, endoplasmic reticulum, and mitochondrial pathways. Six core genes were screened, including RPS7, RPL36A, RPS9, RSL24D1, RPL41, and OSTC, all of which were upregulated under OGD and normalized after reoxygenation. We identified novel ribosome-related genes as potential biomarkers of BBB injury in cerebral infarction. These findings enhance our understanding of BBB pathophysiology and offer new targets for diagnostic and therapeutic strategies in ischemic stroke.
Project description:Oxygen and glucose metabolism plays a pivotal role in many (patho)physiological conditions. In particular, oxygen and glucose deprivation (OGD) occurs during ischemia and stroke, resulting in extensive tissue injury and cell death. We applied time-resolved ribosome profiling technique to assess early events at the level of gene expression in rat pheochromocytoma PC12 cells during short-term OGD. Most substantial alterations in transcripts levels and their translation were seen to occur in the first 20 minutes of OGD. The rapid adaptation of translation apparatus to OGD is global and involves altered elongation and initiation rates. We also observed salient and reproducible alterations in ribosome densities of individual mRNAs such as increased translation of particular upstream Open Reading Frames (uORFs); induced site-specific arrests of the ribosomes and synthesis of extended protein isoforms.
Project description:Mouse Bone marrow-derived macrophages (BMDMs) and HT22 neuronal cells (subjected to 2-hour oxygen-glucose deprivation, OGD-2h) were used in this study. To investigate cell migration, a Transwell assay was performed. For pharmacological intervention, cells were treated with either the serotonin receptor 2B (5-HT2B) agonist BW723C86 or dimethyl sulfoxide (DMSO) as a vehicle control. Following treatment, global protein expression profiling was performed using mass spectrometry-based quantitative proteomics.
Project description:Oxygen-glucose deprivation (OGD) is a cellular phenomenon consistently observed upon occurrence of ischemic stroke which eventually results in neuronal death. Neuronal cell death after ischemia takes place via two distinct processes, necrosis and apoptosis. Apoptosis, programmed cell death, is denoted by chromatin condensation, nuclear blebbing, cellular shrinkage, and DNA fragmentation. Unlike apoptosis, cellular swelling and lysis is suggestive of necrosis. However, these two processes are related not only to the severity but also to the duration of ischemia. Microarray analysis was carried out using 20 Illumina mouse Ref8V1.1 genechip arrays. The assignment of the arrays was as follows: Controls (n=5); exposure to OGD for 10min, 5h, 8h, 15h and 24 h (n=3 respectively).
Project description:Migroglia cells were exposed to oxygen-glucose deprivation (OGD) for 3 h. The miRNA was isolated and the expression profiles of OGD-activated cells were compared with the profiles of resting cells.