Related Experiment Video
Updated: Aug 2, 2026
Isolation and Flow Cytometric Analysis of Immune Cells from the Ischemic Mouse Brain
Published on: February 12, 2016
MicroRNA-4443 regulates monocyte activation by targeting tumor necrosis factor receptor associated factor 4 in
1Department of Neurology, Affiliated ZhongDa Hospital, School of Medicine, Southeast University, Nanjing, China.
Background And Purpose:
MicroRNAs (miRNAs) have been demonstrated to play crucial roles in the early stage of acute ischaemic stroke (AIS). The purpose of this study was to investigate the expression patterns of miRNAs in peripheral blood mononuclear cells (PBMCs) from AIS patients and further explore related molecular mechanisms in stroke-induced immunodeficiency syndrome (SIDS).
Methods:
The miRNA expression patterns of PBMCs were detected by miRNA microarray and validated by quantitative real-time polymerase chain reaction (qRT-PCR) in AIS patients and healthy controls. Bioinformatics methods and luciferase reporter assays were used to detect the downstream target genes. Following stimulation with lipopolysaccharide and interleukin-4, the expression of miR-4443, tumor necrosis factor receptor associated factor 4 (TRAF4) and the nuclear factor kappa B (NF-κB) pathway were evaluated. Furthermore, transfection with miR-4443 mimic or inhibitor in the monocytes was carried out to gain insight into the mechanisms in SIDS.
Results:
Interleukin-10 in AIS patients was significantly higher than that of healthy controls. The miRNA microarray analysis and qRTPCR validation showed that only miR-4443 was upregulated expressed in PBMCs from AIS patients, especially in monocytes. miR-4443 was shown to directly interact with the 3' untranslated regions of TRAF4, resulting in suppression of TRAF4 protein expression. Furthermore, the expression of miR-4443 and TRAF4 was regulated by stimulation with lipopolysaccharide or interleukin-4. Additionally, overexpression of miR-4443 suppressed the TRAF4/Iκα/NF-κB signaling pathway to activate the expression of anti-inflammatory cytokines in monocytes.
Conclusions:
The increased expression of miR-4443 induced monocyte dysfunction by targeting TRAF4, which may function as a crucial mediator in SIDS.
Insights
Increased miR-4443 in acute ischemic stroke (AIS) patients targets TRAF4, causing monocyte dysfunction and contributing to stroke-induced immunodeficiency syndrome (SIDS). This finding highlights miR-4443 as a potential mediator in SIDS.
Area of Science:
- Immunology
- Molecular Biology
- Genetics
Background:
- MicroRNAs (miRNAs) are critical in acute ischemic stroke (AIS) pathogenesis.
- Stroke-induced immunodeficiency syndrome (SIDS) involves complex molecular mechanisms.
- Understanding miRNA expression in AIS is vital for elucidating SIDS.
Purpose of the Study:
- To investigate miRNA expression patterns in peripheral blood mononuclear cells (PBMCs) of AIS patients.
- To explore the molecular mechanisms underlying SIDS.
- To identify specific miRNAs and their targets involved in stroke-induced immune dysfunction.
Main Methods:
- miRNA expression profiling using microarray and quantitative real-time polymerase chain reaction (qRT-PCR).
- Bioinformatics analysis and luciferase reporter assays to identify miRNA targets.
- In vitro studies involving cell stimulation and transfection to elucidate molecular pathways.
Main Results:
- miR-4443 was significantly upregulated in PBMCs of AIS patients, particularly in monocytes.
- miR-4443 directly targets and suppresses tumor necrosis factor receptor associated factor 4 (TRAF4) expression.
- Overexpression of miR-4443 inhibited the TRAF4/IκB/NF-κB pathway, promoting anti-inflammatory cytokine expression.
Conclusions:
- Upregulated miR-4443 in AIS patients induces monocyte dysfunction by targeting TRAF4.
- miR-4443 acts as a key mediator in the development of SIDS.
- Targeting miR-4443 may offer a therapeutic strategy for SIDS.
More Related Videos
Related Concept Videos
01:22MicroRNAs
01:22MicroRNAs