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Published on: March 7, 2022
CircTRHDE knockdown protects WI-38 cells against LPS-induced inflammatory injury
Lifang Sun1, Jingyan Liu1, Xu Sun1
1Department of Pediatrics, Zhengzhou Central Hospital, Zhengzhou University, Zhengzhou, Henan, China.
Background:
Circular RNAs (circRNAs) have been reported to be involved in the progression of infantile pneumonia. Here, we investigated the function of circTRHDE in lipopolysaccharide (LPS)-induced cell inflammatory injury to evaluate its role in infantile pneumonia progression.
Methods:
The circTRHDE, microRNA (miR)-381-3p and TNF-receptor associated factor 3 (TRAF3) expression were detected by quantitative real-time PCR. LPS-induced WI-38 cells were used to construct an inflammatory injury model. Cell viability, inflammation and apoptosis were measured by cell counting kit assay, ELISA assay and flow cytometry. Caspase3 activity, MDA level and SOD activity were analysed to assess cell apoptosis and oxidative stress. Protein levels were determined using western blot analysis. The interaction between miR-381-3p and circTRHDE or TRAF3 was confirmed by dual-luciferase activity assay and RNA pull-down assay.
Results:
CircTRHDE had increased expression in infantile pneumonia patients and LPS-induced WI-38 cells. LPS treatment inhibited WI-38 cell viability while promoting inflammation, apoptosis and oxidative stress. However, knockdown of circTRHDE remitted LPS-triggered WI-38 cell injury. CircTRHDE could sponge miR-381-3p to positively regulate TRAF3 expression. MiR-381-3p suppressed LPS-induced WI-38 cell inflammatory injury, and this effect was revoked by TRAF3 overexpression. Also, LPS-induced WI-38 cell inflammatory injury restrained by circTRHDE knockdown also were reversed by miR-318-3p inhibitor or TRAF3 overexpression.
Conclusion:
Our findings demonstrated that circTRHDE might be a target for infantile pneumonia treatment, which relieved LPS-induced cell inflammatory injury by the regulation of the miR-318-3p/TRAF3 axis.
Insights
Circular RNA circTRHDE exacerbates infantile pneumonia by promoting inflammation and apoptosis. Targeting circTRHDE may offer a novel therapeutic strategy for infantile pneumonia by modulating the miR-381-3p/TRAF3 pathway.
Area of Science:
- Molecular Biology
- Immunology
- Cell Biology
Background:
- Circular RNAs (circRNAs) are implicated in infantile pneumonia pathogenesis.
- This study investigates the role of circTRHDE in lipopolysaccharide (LPS)-induced inflammatory injury in the context of infantile pneumonia.
Purpose of the Study:
- To elucidate the function of circTRHDE in LPS-induced cell inflammatory injury.
- To evaluate the potential of circTRHDE as a therapeutic target for infantile pneumonia.
Main Methods:
- Quantitative real-time PCR to measure circTRHDE, miR-381-3p, and TRAF3 expression.
- Cell viability, inflammation, apoptosis, and oxidative stress assays in LPS-induced WI-38 cells.
- Western blot, dual-luciferase, and RNA pull-down assays to confirm molecular interactions.
Main Results:
- CircTRHDE expression was elevated in infantile pneumonia patients and LPS-treated cells.
- Knockdown of circTRHDE alleviated LPS-induced inflammatory injury, apoptosis, and oxidative stress.
- CircTRHDE positively regulated TRAF3 expression by sponging miR-381-3p; miR-381-3p suppressed inflammation, an effect reversed by TRAF3 overexpression.
Conclusions:
- CircTRHDE promotes LPS-induced cell inflammatory injury via the miR-381-3p/TRAF3 axis.
- CircTRHDE may serve as a potential therapeutic target for infantile pneumonia.
