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Updated: Jan 15, 2026
Pseudomonas aeruginosa Induced Lung Injury Model
Published on: October 29, 2014
Circ-PRKCI attenuates acute lung injury secondary to sepsis by targeting the miR-106b-5p/GAB1 axis
Yingjie Yin1, Shouqing Zhang2, Jiebing Zheng3
1Department of Critical Care Medicine, The First Affiliated Hospital of Ningbo University, Ningbo, 315010, Zhejiang, China. fyyinyingjie@nbu.edu.cn.
Background:
Acute lung injury (ALI) secondary to sepsis increases the risk of mortality in septic patients. However, the molecular mechanism of alveolar epithelial cell damage in ALI secondary to sepsis remains incompletely understood. Studies demonstrate that circular RNA (circRNA) modulates the progression of ALI secondary to sepsis. This study aims to explore the function and mechanisms of circ-PRKCI in ALI associated with sepsis.
Methods:
The in vitro ALI model was constructed by treating MLE-12 cells with LPS. The expression of circ-PRKCI, miR-106b-5p, GRB2-associated binder 1 (GAB1), and inflammatory cytokines was measured via RT-qPCR. The expression of apoptosis-related proteins and GAB1 protein was analyzed via Western blot. The subcellular localization of circ-PRKCI was determined using immunofluorescence. Cell viability was measured by the CCK-8 assay, and apoptosis was evaluated using flow cytometry. ELISA was employed to measure pro-inflammatory cytokine concentrations in cells and lung tissue samples. The content of oxidative stress markers was measured utilizing corresponding assay kits. The binding relationship between miR-106b-5p and circ-PRKCI or GAB1 was validated through dual-luciferase reporter assays and RNA RIP experiments. The in vivo ALI model was constructed via cecal ligation and puncture (CLP) surgery. HE staining and the W/D ratio test were performed on mouse lung tissues to evaluate the extent of lung injury.
Results:
The findings revealed that circ-PRKCI attenuated apoptosis, inflammation, and oxidative stress in ALI-induced cells, while also suppressing pulmonary inflammation, edema, and apoptotic responses in the murine ALI model. However, miR-106b-5p was capable of reversing the function of circ-PRKCI. Mechanistically, circ-PRKCI modulated the expression of GAB1 by competitively binding to miR-106b-5p.
Conclusion:
Our findings indicated that circ-PRKCI attenuates ALI secondary to sepsis by targeting the miR-106b-5p/GAB1 axis, suggesting that circ-PRKCI holds potential as a therapeutic target for ALI secondary to sepsis.
Insights
Circular RNA PRKCI (circ-PRKCI) reduces acute lung injury (ALI) in sepsis by inhibiting inflammation and apoptosis. It targets the miR-106b-5p/GAB1 pathway, offering a potential therapeutic strategy for sepsis-induced ALI.
Area of Science:
- Molecular Biology
- Immunology
- Cell Biology
Background:
- Sepsis-induced acute lung injury (ALI) is a major cause of mortality.
- The precise molecular mechanisms of alveolar epithelial cell damage in sepsis-ALI are not fully understood.
- Circular RNAs (circRNAs) are implicated in the progression of sepsis-ALI.
Purpose of the Study:
- To investigate the functional role and underlying mechanisms of circ-PRKCI in sepsis-associated ALI.
- To explore circ-PRKCI's interaction with microRNA-106b-5p (miR-106b-5p) and GRB2-associated binder 1 (GAB1).
Main Methods:
- In vitro models using MLE-12 cells treated with lipopolysaccharide (LPS) to mimic ALI.
- In vivo models of ALI induced by cecal ligation and puncture (CLP) in mice.
- Assays included RT-qPCR, Western blot, CCK-8, flow cytometry, ELISA, dual-luciferase reporter assays, and RNA RIP experiments.
Main Results:
- Circ-PRKCI significantly attenuated apoptosis, inflammation, and oxidative stress in both in vitro and in vivo ALI models.
- MiR-106b-5p reversed the protective effects of circ-PRKCI.
- Circ-PRKCI competitively bound to miR-106b-5p, thereby modulating GAB1 expression.
Conclusions:
- Circ-PRKCI mitigates sepsis-induced ALI by targeting the miR-106b-5p/GAB1 axis.
- Circ-PRKCI demonstrates potential as a novel therapeutic target for ALI secondary to sepsis.
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