Related Experiment Video
Updated: Sep 5, 2025
Characterization and Isolation of Mouse Primary Microglia by Density Gradient Centrifugation
Published on: February 16, 2018
Transcriptome sequencing analysis revealed the molecular mechanism of podoplanin neutralization inhibiting
Shuang Qian1,2, Lei Qian3, Ye Yang1,2
1Jiangsu Institute of Hematology, Key Laboratory of Thrombosis and Hemostasis of the Ministry of Health, The First Affiliated Hospital of Soochow University, Suzhou, China.
Background:
Anti-podoplanin antibody (α-PDPN, clone 8.1.1) reduces microglia-mediated inflammation and decreases cerebral infarct volume in mice with stroke. However, the molecular mechanism by which this occurs is unknown. This study sought to systematically analyze the molecular mechanism of α-PDPN treatment on ischemia/reperfusion (I/R)-injured microglia.
Methods:
Microglia BV2 cells were pre-cultured with α-PDPN and then exposed to oxygen-glucose deprivation and reoxygenation (OGD-R) insult. The differentially expressed genes (DEGs) underwent a transcriptome sequencing technology analysis, followed by Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses. Quantitative real-time polymerase chain reaction (PCR) was performed to confirm the transcriptional expression of some DEGs.
Results:
The results showed that α-PDPN downregulated 338 genes and upregulated 340 genes in the BV2 cells. The GO items of the downregulated DEGs mainly involved biological processes, such as the response to the interferon (IFN), lipopolysaccharide-mediated signaling pathway, and the regulation of cell chemotaxis and migration. The upregulated molecular function mainly involved glucocorticoid-receptor binding. Further, the KEGG pathway analysis indicated that the enriched categories for the upregulated DEGs mainly involved the adenosine triphosphate (ATP) binding cassette transporters. However, the interleukin-17 signaling pathway, IFN signaling pathway, tumor necrosis factor signaling pathway, transforming growth factor beta (TGF-ꞵ) signaling pathway, nucleotide-binding and oligomerization domain (NOD)-like receptor signaling pathway, cytokine-cytokine receptor interaction, and chemokine signaling pathway were downregulated by the α-PDPN treatment.
Conclusions:
Numerous inflammation-related signaling pathways were regulated by the α-PDPN treatment in the OGD-R injured BV2 cells. This study provided further insights into the protective mechanism of α-PDPN treatment in ischemic stroke.
Insights
Anti-podoplanin antibody treatment reduces inflammation and brain damage after stroke by altering gene expression in microglia. This study reveals key molecular pathways involved in this protective effect, offering insights into stroke treatment strategies.
Area of Science:
- Neuroscience
- Immunology
- Molecular Biology
Background:
- Anti-podoplanin antibody (α-PDPN) shows promise in reducing stroke-related brain damage and inflammation.
- The precise molecular mechanisms underlying α-PDPN's protective effects on microglia remain unclear.
Purpose of the Study:
- To systematically investigate the molecular mechanisms of α-PDPN in protecting against ischemia/reperfusion (I/R)-induced microglial injury.
- To identify key signaling pathways and gene expression changes modulated by α-PDPN in microglia.
Main Methods:
- Transcriptome sequencing analyzed gene expression changes in BV2 microglia treated with α-PDPN and subjected to oxygen-glucose deprivation/reoxygenation (OGD-R).
- Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses identified enriched biological processes and pathways.
- Quantitative real-time PCR validated the expression of selected differentially expressed genes (DEGs).
Main Results:
- α-PDPN significantly altered gene expression, downregulating 338 genes and upregulating 340 genes in OGD-R-injured microglia.
- Downregulated DEGs were associated with interferon responses, lipopolysaccharide signaling, and cell migration.
- Upregulated DEGs were linked to glucocorticoid-receptor binding and ATP binding cassette transporters, while key inflammatory pathways (e.g., IL-17, TNF, TGF-β) were downregulated.
Conclusions:
- α-PDPN treatment modulates numerous inflammation-related signaling pathways in microglia following ischemic injury.
- This study elucidates the molecular underpinnings of α-PDPN's neuroprotective effects in ischemic stroke.
- Findings provide a foundation for developing targeted therapies for stroke based on α-PDPN's mechanism of action.
More Related Videos
08:23Isolation and Flow Cytometric Assessment of Neuroimmune Interactions in a Mini-Stroke Murine Model
Published on: June 20, 2025
07:15Mechanism of Kemeng Fang's Inhibition of Podocyte Apoptosis in Rats with Membranous Nephropathy through the PI3K/AKT Signaling Pathway
Published on: August 23, 2024