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Updated: Jul 23, 2025
Bronchoalveolar Lavage Exosomes in Lipopolysaccharide-induced Septic Lung Injury
Published on: May 21, 2018
Molecular mechanism of miRNA-23a in sepsis-induced lung injury
Pingli Zhang1, Qikun Huang1, Peiyuan Liao1
1Department of Pediatric, Qilu Hospital (Qingdao), Cheeloo College of Medicine, Shandong University Qingdao 266035, Shandong, China.
Objective:
MicroRNA-23a-3p (miR-23a) is a novel gene regulator involved in inflammation. This study aimed to explore the molecular mechanism of miR-23a in sepsis-induced lung injury both in vitro and in vivo.
Methods:
Lipopolysaccharide (LPS)- and ATP-stimulated human myeloid leukemia mononuclear cells (THP-1) and Human Bronchial Epithelial Cells (BEAS-2B) cell lines were used, while cecal ligation and puncture (CLP)-induced sepsis BABL/c mice were constructed. The mRNA expression levels of interleukin (IL)-18, IL-1β, and miR-23a were determined, and Western blotting was used to measure CXCR4/PTEN/PI3K/AKT signaling. The concentrations of cytokines and Nod-like receptor family pyrin domain-containing 3 (NLRP3) were determined using an enzyme-linked immunosorbent assay. Lung tissue of mice was subjected to hematoxylin-eosin staining for examining myocardial injury.
Results:
MiR-23a inhibited NLRP3 inflammasome activation in LPS- and ATP-stimulated THP-1 and BEAS-2B cells (P<0.05). Overexpression of miR-23a decreased the lactate dehydrogenase release rate in the cells (P<0.05). Meanwhile, miR-23a overexpression decreased the concentration and gene expression of IL-1β and IL-18 in CXCR4 positive cells (P<0.05). Conversely, miR-23a knockdown increased the concentration and gene expression of IL-1β and IL-18 (P<0.05). Additionally, PTEN and p53 proteins were up-regulated in miR-23a mimic group and down-regulated in miR-23a inhibitor group (P<0.05). Furthermore, miR-23a expression was decreased in sepsis-induced lung injury mice (P<0.05). MiR-23a overexpression reduced the sepsis-induced lung injury probably by inhibiting acetylcholinesterase activity and expression levels of IL-1β, IL-18, capase-1, and NLRP3 (P<0.05).
Conclusion:
miR-23a can significantly alleviate sepsis-induced lung injury in CLP-induced septic mice and LPS-stimulated cell lines by suppressing NLRP3 inflammasome activation and inflammatory response, while promoting the CXCR4/PTEN/PI3K/AKT pathway.
Insights
MicroRNA-23a-3p (miR-23a) mitigates sepsis-induced lung injury by inhibiting the NLRP3 inflammasome and inflammatory response. This study reveals miR-23a
Area of Science:
- Molecular Biology
- Immunology
- Pathology
Background:
- MicroRNA-23a-3p (miR-23a) is recognized as a key regulator of inflammatory processes.
- Sepsis-induced lung injury (ALI) is a severe condition characterized by excessive inflammation.
- Understanding the role of microRNAs in ALI pathogenesis is crucial for developing targeted therapies.
Purpose of the Study:
- To elucidate the molecular mechanisms underlying the function of miR-23a in sepsis-induced ALI.
- To investigate the impact of miR-23a on the NLRP3 inflammasome pathway and inflammatory cytokine production.
- To evaluate the therapeutic potential of miR-23a in both in vitro and in vivo models of ALI.
Main Methods:
- Utilized lipopolysaccharide (LPS)- and ATP-stimulated THP-1 and BEAS-2B cell lines for in vitro studies.
- Established a cecal ligation and puncture (CLP)-induced sepsis model in BABL/c mice for in vivo investigations.
- Quantified mRNA and protein expression of key inflammatory mediators (IL-1β, IL-18, NLRP3) and signaling pathway components (CXCR4, PTEN, PI3K, AKT) using RT-qPCR and Western blotting.
- Assessed lung injury through hematoxylin-eosin staining and measured cytokine levels via ELISA.
Main Results:
- miR-23a overexpression significantly inhibited NLRP3 inflammasome activation and reduced IL-1β and IL-18 levels in stimulated cells.
- miR-23a knockdown led to increased IL-1β and IL-18 expression, exacerbating inflammatory responses.
- In CLP-induced sepsis mice, miR-23a expression was downregulated, and its overexpression ameliorated lung injury by suppressing inflammatory markers and NLRP3 inflammasome activation.
- miR-23a promoted the CXCR4/PTEN/PI3K/AKT pathway, suggesting a protective role.
Conclusions:
- miR-23a plays a critical protective role in sepsis-induced lung injury.
- The therapeutic effects of miR-23a involve the suppression of NLRP3 inflammasome activation and inflammatory responses.
- Modulation of the CXCR4/PTEN/PI3K/AKT pathway by miR-23a contributes to its protective function in ALI.
