Quercetin attenuated necroptosis and apoptosis caused by LPS-induced mitochondrial function dysfunction through the METTL3-mediated PTEN m6A methylation/PI3K/AKT signaling in broiler livers

Abstract

Background

Quercetin (QUE), a natural flavonoid, offered an efficient protection against organism injury. N6-methyladenosine (m⁶A) methylation is considered to be the most prevalent and abundant modifications involved in various diseases.

Purpose

We sought to explore protective roles of QUE in mitigating necroptosis and apoptosis triggered by LPS-induced imbalances in mitochondria dynamic and energy metabolism in broiler livers, with a focus on m⁶A methylation modulation.

Study design/Methods

We used LPS as a stimulus and treated with QUE to establish this in vivo and in vitro. In addition, we treated LMH cells with siMETTL3 (80 nM) to determine its detailed mechanism.

Results

Our findings revealed QUE significantly decreased METTL3 expression, leading to a decrease in PTEN m⁶A methylation and factors related to mitochondria fission, necroptosis, and apoptosis in the QUE+LPS group. In contrast, QUE treatment promoted the expression levels of marker factors for mitochondria fusion, energy metabolism, anti-apoptosis, and PI3K/AKT compared with the LPS group. Additionally, an increase of ΔΨm, ATP content, and ATPase activity was observed. AO/EB staining, Flow cytometry and TUNEL assays confirmed QUE inhibited LPS-induced apoptosis and necroptosis. Molecular docking analysis and cellular thermal shift assay supported an interaction between QUE and METTL3.

Conclusion

In summary, QUE mitigated necroptosis and apoptosis triggered by LPS-induced disorders of mitochondrial kinetic and metabolic processes in broiler livers through its interaction with METTL3, regulating PTEN m⁶A methylation/PI3K/AKT signaling pathway. This study enhances our understanding of biological functions for QUE and lays a theoretical foundation for developing new therapeutic interventions, highlighting its potential value.