KLF14 restrained NLRP3-mediated pyroptosis in sepsis-induced acute lung injury by activating DTX3L.

INTRODUCTION

Acute lung injury (ALI) is characterized by inflammation of the lungs, leading to impaired gas exchange and respiratory distress. This study aimed to investigate the mechanisms underlying the role of Krüppel-like factor 14 (KLF14) in ALI pathogenesis.

MATERIAL AND

METHODS

Protein and gene expression levels were quantified using quantitative reverse transcription polymerase chain reaction (qRT-PCR) and western blot. Cell viability and pyroptosis were assessed by Cell Counting Kit-8 (CCK-8) and flow cytometry. Interleukin (IL)-18 and IL-1 β levels were measured by enzyme-linked immunosorbent assay (ELISA), while lactate dehydrogenase (LDH) activity was evaluated with a commercial assay kit. The interaction between Deltex E3 ubiquitin ligase 3-like (DTX3L) and KLF14 or nucleotide-binding domain like receptor 3 (NLRP3) was analyzed using chromatin immunoprecipitation (ChIP), co-immunoprecipitation (Co-IP), and dual-luciferase reporter assay.

RESULTS

BEAS-2B cells treated with lipopolysaccharide (LPS) exhibited reduced viability and elevated pyroptosis-related markers. Notably, KLF14 upregulation suppressed NLRP3-mediated pyroptosis in LPS-induced cells. Similarly, DTX3L overexpression attenuated pyroptosis in LPS-treated BEAS-2B cells. Mechanistically, KLF14 enhanced DTX3L transcription, and DTX3L promoted NLRP3 degradation via ubiquitination. Furthermore, KLF14 upregulation inhibited NLRP3-driven pyroptosis by inducing DTX3L expression.

CONCLUSIONS

Upregulation of KLF14 inhibited NLRP3-mediated pyroptosis through DTX3L activation, thereby improving sepsis-induced acute lung injury.