Circ-Bnc2 alleviates neuroinflammation in LPS-stimulated microglial cells to inhibit neuron cell apoptosis through regulating miR-497a-5p/HECTD1 axis

Background: Neuroinflammation caused by microglia cells activation and the apoptosis of neuron cells are associated with the occurrence of depression. Circ-Bnc2 has been shown to be significantly downregulated in depression mice, but its role in the progression of depression remains unclear.

Methods: Lipopolysaccharide (LPS) was used to treat BV2 microglial cells to induce neuroinflammation. The expression of circ-Bnc2, microRNA (miR)-497a-5p, and HECT domain E3 ubiquitin protein ligase 1 (HECTD1) was measured by quantitative real-time PCR. The protein levels of neuroinflammation markers, apoptosis markers, and HECTD1 were determined by western blot analysis. ELISA assay was used to examine the concentrations of inflammatory factors. After HT22 cells were cultured with the conditioned medium of LPS-induced BV2 cells, the proliferation and apoptosis of HT22 cells were assessed by cell counting kit 8 assay, EdU assay, and flow cytometry. In addition, the interaction between miR-497a-5p and circ-Bnc2 or HECTD1 was confirmed by dual-luciferase reporter assay, RIP assay, and RNA pull-down assay.

Results: Our data showed that circ-Bnc2 was lowly expressed in LPS-induced BV2 cells. Function experiments suggested that circ-Bnc2 could inhibit LPS-induced neuroinflammation in BV2 cells to repress HT22 cell apoptosis and promote proliferation. Circ-Bnc2 could sponge miR-497a-5p, and the neuroprotective function of circ-Bnc2 could be reversed by miR-497a-5p overexpression. Additionally, miR-497a-5p could target HECTD1. miR-497a-5p inhibitor could alleviate LPS-induced neuroinflammation in BV2 cells and reduce HT22 cell apoptosis, which also could be reversed by HECTD1 knockdown. Moreover, circ-Bnc2 had a positive regulation on HECTD1 expression by sponging miR-497a-5p.

Conclusion: In summary, our results confirmed that circ-Bnc2 could inhibit neuroinflammation and neuron cell apoptosis by regulating miR-497a-5p/HECTD1 axis, suggesting that circ-Bnc2 might be a potential target for depression treatment.