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Updated: May 23, 2026

Analysis of Combinatorial miRNA Treatments to Regulate Cell Cycle and Angiogenesis
Published on: March 30, 2019
miR-146a, an IL-1β responsive miRNA, induces vascular endothelial growth factor and chondrocyte apoptosis by
Jing Li1, Jingang Huang, Liming Dai
1Key Laboratory of Stem Cell Biology, Institute of Health Sciences, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences & Shanghai Jiao Tong University School of Medicine, China, 200025.
Introduction:
miR-146a is one of the first identified miRNAs expressed differentially in osteoarthritis (OA) cartilage. However, the role it plays in OA pathogenesis is not clear. The aim of this study is to identify a molecular target of miR-146a, thereby elucidating its function in chondrocytes during OA pathogenesis.
Methods:
Primary chondrocytes from Sprague-Dawley rats were treated with IL-1β before the expression levels of miR-146a, Smad4 and vascular endothelial growth factor (VEGF) were quantified by real-time PCR and/or western blotting. The effect of miR-146a on cellular response to transforming growth factor (TGF)-β1 was quantified by a luciferase reporter harboring TGF-β1 responsive elements and by extracellular signal-regulated kinase assay. The effect of miR-146a on apoptosis was quantified by the TUNEL assay. OA pathogenesis was surgically induced with joint instability in rats, evaluated by histopathological analysis with safranin O staining, and the expression levels of miR-146a, Smad4, and VEGF were quantified using real-time PCR and/or immunohistochemistry.
Results:
IL-1β treatment of chondrocytes increased the expression levels of miR-146a and VEGF and decreased the levels of Smad4 in a time-dependent manner. miR-146a upregulated VEGF expression and downregulated Smad4 expression in chondrocytes, while a miR-146a inhibitor acted in a converse manner. Smad4, a common mediator of the TGF-β pathway, is identified as a direct target of miR-146a by harboring a miR-146a binding sequence in the 3'-UTR region of its mRNA. Mutation of the binding sequence significantly relieved the inhibition of the Smad4 reporter activity by miR-146a. Furthermore, miR-146a upregulation of VEGF is mediated by Smad4. Expression of miR-146a led to a reduction of cellular responsiveness to TGF-β and an increase of apoptosis rate in chondrocytes. In vivo, cartilage from surgically induced OA rats displayed higher levels of miR-146a and VEGF compared with the sham group. In contrast, Smad4 expression level was lower in the OA group than the sham group.
Conclusion:
IL-1β responsive miR-146a is overexpressed in an experimentally induced OA model, accompanied by upregulation of VEGF and downregulation of Smad4 in vivo. miR-146a may contribute to OA pathogenesis by increasing VEGF levels and by impairing the TGF-β signaling pathway through targeted inhibition of Smad4 in cartilage.
Insights
Osteoarthritis (OA) involves increased miR-146a, which upregulates VEGF and downregulates Smad4, impairing TGF-β signaling. This microRNA contributes to OA pathogenesis by promoting cartilage degradation and apoptosis.
Area of Science:
- Molecular Biology
- Biochemistry
- Pathology
Background:
- Osteoarthritis (OA) is characterized by differential expression of microRNAs (miRNAs), including miR-146a, in cartilage.
- The precise role of miR-146a in the pathogenesis of OA remains unclear.
- Identifying miR-146a targets is crucial for understanding its function in chondrocytes during OA development.
Purpose of the Study:
- To identify a molecular target of miR-146a.
- To elucidate the function of miR-146a in chondrocytes during OA pathogenesis.
- To investigate the role of miR-146a in the transforming growth factor (TGF)-β signaling pathway and apoptosis.
Main Methods:
- Primary chondrocytes and surgically induced OA rat models were utilized.
- Expression levels of miR-146a, Smad4, and vascular endothelial growth factor (VEGF) were quantified using real-time PCR and western blotting/immunohistochemistry.
- Luciferase reporter assays, extracellular signal-regulated kinase (ERK) assays, and TUNEL assays were performed to assess miR-146a's effects on TGF-β signaling and apoptosis.
Main Results:
- Interleukin-1 beta (IL-1β) treatment increased miR-146a and VEGF, while decreasing Smad4 in chondrocytes.
- Smad4 was identified as a direct target of miR-146a, mediating miR-146a's upregulation of VEGF.
- miR-146a expression reduced chondrocyte responsiveness to TGF-β1 and increased apoptosis; OA cartilage showed elevated miR-146a and VEGF, with decreased Smad4.
Conclusions:
- miR-146a is overexpressed in an experimental OA model and is responsive to IL-1β.
- miR-146a contributes to OA pathogenesis by upregulating VEGF and inhibiting the TGF-β pathway via Smad4 targeting.
- Targeting miR-146a may offer a therapeutic strategy for OA.
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