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miR-374b-5p is increased in deep vein thrombosis and negatively targets IL-10
Yunhong Zhang1, Xiuming Miao2, Zhen Zhang3
1School of Medicine and Life Sciences, University of Jinan-Shandong Academy of Medical Sciences, 18877 Jingshi Road, Jinan 250062, Shandong, China; Laboratory for Molecular Immunology, Institute of Basic Medicine, Shandong Provincial Hospital Affiliated to Shandong First Medical University, 18877 Jingshi Road, Jinan 250062, Shandong, China.
Insights
Increased miR-374b-5p downregulates interleukin-10 (IL-10) expression, promoting deep venous thrombosis (DVT) formation. This microRNA presents a potential diagnostic marker and therapeutic target for DVT.
Area of Science:
- Molecular Biology
- Cardiovascular Research
- Epigenetics
Background:
- Deep venous thrombosis (DVT) is a significant cardiovascular complication.
- Decreased interleukin-10 (IL-10) levels are implicated in DVT pathogenesis.
- The precise molecular mechanisms behind IL-10 downregulation in DVT remain unclear.
Purpose of the Study:
- To investigate the role of post-transcriptional epigenetic modifications in IL-10 regulation in DVT.
- To identify specific microRNAs (miRNAs) involved in controlling IL-10 expression in DVT.
- To explore the therapeutic potential of targeting these molecular interactions in DVT.
Main Methods:
- miRNA expression profiling in DVT patients using microarray analysis.
- In silico prediction and experimental validation of miRNA targets for IL-10.
- Quantitative real-time PCR (qRT-PCR) and enzyme-linked immunoassay for IL-10 and miR-374b-5p.
- Dual luciferase reporter assays to confirm direct miRNA-mRNA interaction.
- Murine DVT models to assess the in vivo effects of miR-374b-5p modulation.
Main Results:
- IL-10 expression was significantly decreased, while miR-374b-5p levels were elevated in DVT patients.
- A strong negative correlation was observed between miR-374b-5p and IL-10 expression.
- Overexpression of miR-374b-5p reduced IL-10, whereas its inhibition increased IL-10 levels.
- In vivo studies confirmed that elevated miR-374b-5p aggravated DVT, while its inhibition ameliorated the condition.
- Direct binding of miR-374b-5p to the IL10 gene was experimentally verified.
Conclusions:
- Elevated miR-374b-5p promotes DVT by suppressing IL-10 expression.
- miR-374b-5p represents a promising diagnostic biomarker for DVT.
- Targeting miR-374b-5p offers a potential therapeutic strategy for managing DVT.
Background:
Deep venous thrombosis (DVT) is one of the most common venous thromboembolic (VTE) disorders and the third leading cardiovascular complication. Accumulating evidence has shown that decreased interleukin-10 (IL-10) was involved in DVT. However, the underlying molecular mechanisms are still largely unknown. Here, we proposed that the epigenetic modification of IL-10 at the post-transcriptional level may be a crucial trigger for IL-10 down-regulation in DVT.
Methods:
miRNA expression in DVT was profiled by miRNA microarray analysis. The upstream miRNA regulators of IL-10 were predicted by in silico target prediction tools. The expression of IL-10 mRNA and miR-374b-5p were examined by quantitative real-time PCR (qRT-PCR) and the protein expression of IL-10 was detected by enzyme-linked immunoassay. Dual luciferase reporter assay was used to identify the interaction between miR-374b-5p and IL10. A murine model of DVT was developed and the localization of miR-374b-5p was visualized in vitro by fluorescence in situ hybridization. The biological effects of miR-374b-5p on IL-10 was examined both in vitro and in vivo.
Results:
Microarray and qRT-PCR results showed that the IL-10 expression was decreased while miR-374b-5p level was increased substantially in peripheral blood mononuclear cells of DVT patients, and there was significant negative correlation between miR-374b-5p and IL-10. Experiments in vitro showed that overexpressed miR-374b-5p reduced IL-10 expression, while miR-374b-5p knockdown increased IL-10 expression. Moreover, in vivo studies revealed that DVT mice with anti-IL-10 antibody or agomiR-374b-5p delivery resulted in decreased IL-10 expression and aggravated DVT formation, whereas antagomiR-374b-5p acted inversely. Dual luciferase reporter assay identified direct binding between miR-374b-5p and IL10.
Conclusions:
These findings suggest that increased miR-374b-5p promotes DVT formation by downregulating IL-10 expression. miR-374b-5p may be explored as a promising diagnostic marker and therapeutic target for DVT.
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