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Downregulation of circ_0035292 Alleviates LPS-Induced WI-38 Cell Injury via Targeting miR-494-3p/TLR4 Pathway in
Zhenzhao Dai1, Jiansheng Hu2, Zhiying Luo2
1Department of Pediatrics, Affiliated Hospital of Jinggangshan University, Ji'an, Jiangxi, China.
Abstract:
Circular RNAs (circRNAs) have been confirmed to mediate infantile pneumonia development. In this, we investigated the role and new mechanism of circ_0035292 regulating infantile pneumonia progression. Lipopolysaccharide (LPS)-treated WI-38 cells were used to mimic infantile pneumonia cell injury models. Quantitative real-time PCR was used to measure circ_0035292, microRNA (miR)-494-3p and toll-like receptor 4 (TLR4). Cell proliferation and apoptosis were assessed by MTT assay, EdU assay, and flow cytometry. Protein expression was tested using western blot analysis. Inflammation and oxidative stress were evaluated by measuring IL-6, IL-1β, MDA and SOD levels using ELISA assay and corresponding kits. RNA interaction was confirmed by dual-luciferase reporter assay and RIP assay. Circ_0035292 had elevated expression in infantile pneumonia patients and LPS-induced WI-38 cells. Silenced circ_0035292 could enhance WI-38 cell proliferation, while suppress apoptosis, inflammation and oxidative stress under LPS treatment. Mechanically, circ_0035292 targeted miR-494-3p to positively regulate TLR4. The rescue experiments indicated that miR-494-3p inhibitor abolished the function of circ_0035292 knockdown, and TLR4 overexpression reversed the inhibitory effect of miR-494-3p on LPS-induced WI-38 cell injury. Circ_0035292 might be a potential target for infantile pneumonia treatment, which knockdown could relieve LPS-induced cell injury via the regulation of miR-494-3p/TLR4 axis.
Insights
Circular RNA circ_0035292 promotes infantile pneumonia by increasing inflammation and oxidative stress. Silencing circ_0035292 may offer a therapeutic strategy for infantile pneumonia by targeting the miR-494-3p/TLR4 pathway.
Area of Science:
- Molecular Biology
- Cell Biology
- Immunology
Background:
- Circular RNAs (circRNAs) play a role in infantile pneumonia.
- Understanding the specific mechanisms of circRNAs in infantile pneumonia is crucial for developing effective treatments.
Purpose of the Study:
- To investigate the role and mechanism of circ_0035292 in infantile pneumonia progression.
- To explore the potential of circ_0035292 as a therapeutic target.
Main Methods:
- Used lipopolysaccharide (LPS)-induced WI-38 cells as an infantile pneumonia model.
- Quantified gene expression (circ_0035292, miR-494-3p, TLR4) using qRT-PCR.
- Assessed cell proliferation, apoptosis, inflammation, and oxidative stress markers.
- Confirmed RNA interactions using dual-luciferase reporter and RIP assays.
Main Results:
- Circ_0035292 expression was elevated in infantile pneumonia patients and LPS-treated cells.
- Silencing circ_0035292 reduced inflammation and oxidative stress while enhancing cell proliferation and suppressing apoptosis.
- Circ_0035292 targets miR-494-3p to positively regulate toll-like receptor 4 (TLR4).
- Knockdown of circ_0035292 alleviated LPS-induced cell injury via the miR-494-3p/TLR4 axis.
Conclusions:
- Circ_0035292 acts as a key regulator in infantile pneumonia.
- The circ_0035292/miR-494-3p/TLR4 axis is a potential therapeutic target for infantile pneumonia.
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