Lipopolysaccharide-induced lung cell inflammation and apoptosis are enhanced by circ_0003420/miR-424-5p/TLR4 axis via inactivating the NF-κB signaling pathway.

BACKGROUND

Circular RNAs (circRNAs) can regulate disease progression, including sepsis-induced acute lung injury (ALI). This research aimed at investigating the function of circ_0003420 in lipopolysaccharide (LPS)-treated lung cells, as well as the functional mechanism.

METHODS

Enzyme-linked immunosorbent assay was used for inflammation analysis. Cell viability and proliferation were examined using Cell Counting Kit-8 assay and EdU assay. Cell apoptosis was measured by flow cytometry. Western blot was used for protein detection. Reverse transcription-quantitative polymerase chain reaction assay was performed for quantification of circ_0003420, microRNA-424-5p (miR-424-5p) or toll-like receptor (TLR4). The interaction between miR-424-5p and circ_0003420 or TLR4 was conducted through dual-luciferase reporter assay and RNA immunoprecipitation (RIP) assay.

RESULTS

Lung cell inflammation and apoptosis were promoted, but cell viability and proliferation were inhibited by LPS. Silence of circ_0003420 attenuated the LPS-mediated lung cell injury. Circ_0003420 could interact with miR-424-5p. The protective function by knockdown of si-circ_0003420 was relieved by miR-424-5p inhibition in LPS-treated cells. TLR4 served as a downstream target of miR-424-5p. Overexpression of miR-424-5p repressed inflammatory and apoptotic damages in LPS-treated lung cells via downregulating TLR4. Circ_0003420 upregulated the TLR4 level by targeting miR-424-5p and circ_0003420 regulated the NF-κB signaling pathway through the miR-424-5p/TLR4 axis.

CONCLUSION

These results uncovered that circ_0003420 contributed to the LPS-induced lung cell injury via activating the miR-424-5p/TLR4-related NF-κB signaling pathway. Circ_0003420 might be a therapeutic target in sepsis-induced ALI.