Related Experiment Video
Updated: Feb 11, 2026
Bronchoalveolar Lavage Exosomes in Lipopolysaccharide-induced Septic Lung Injury
Published on: May 21, 2018
MicroRNA-326 aggravates acute lung injury in septic shock by mediating the NF-κB signaling pathway
Chun-Ting Wu1, Yan Huang1, Zhen-Ye Pei1
1Department of Pulmonary and Critical Care Medicine, Beijing Anzhen Hospital, Capital Medical University, Beijing Institute of Heart, Lung and Blood Vessel Diseases, Beijing, 100029, PR China.
Abstract:
Our previous studies have demonstrated that the activation of the nuclear factor-kappa B (NF-κB) signaling pathway contributes to the development of lipopolysaccharide (LPS)-induced acute lung injury (ALI) as well as an inflammatory reaction, and its inhibition may provide future therapeutic values. Thereby, this study aims to explore the effects of miR-326 on inflammatory response and ALI in mice with septic shock via the NF-κB signaling pathway. The study included normal mice and LPS-induced mouse models of septic shock with ALI. Modeled mice were transfected with the blank plasmid, miR-326 mimic, miR-326 inhibitor, si-BCL2A1 and miR-326 inhibitor + si-BCL2A1. Mean arterial pressure (MAP), airway pressure (AP), heart rate (HR) and lung wet dry (W/D) ratio were determined. Serum levels of interleukin (IL)-6, IL-10, IL-1β, and tumor necrosis factor-α (TNF-α) were detected using ELISA. Reverse transcription quantitative polymerase chain reaction (RT-qPCR) and Western blot analysis were performed to detect the miR-326 expression and expression levels of BCL2A1, related genes of inflammatory response and the NF-κB signaling pathway in lung tissues. Cell viability and apoptosis were measured using the CCK-8 assay and flow cytometry, respectively. Compared to the ALI models and those transfected with blank plasmid, the up-regulated miR-326 expression and silenced BCL2A1 lead to decreased levels of MAP, increased AP, HR and lung W/D, increased serum levels of IL-6, IL-10, IL-1β and TNF-α, increased expressions of IL-6, IL-1β, TNF-α, NF-κB p65 (p-NF-κB p65), and iNOS with decreased expressions of BCL2A1s as well as inhibition of cell viability and enhanced cell apoptosis; the down-regulated miR-326 expression reversed the aforementioned situation. MiR-326 targeting the BCL2A1 gene activated the NF-κB signaling pathway, resulting in aggravated inflammatory response and lung injury of septic shock with ALI in mice.
Insights
MicroRNA-326 (miR-326) exacerbates lipopolysaccharide-induced acute lung injury (ALI) by targeting BCL2A1 and activating the NF-κB pathway. Inhibiting miR-326 may offer a therapeutic strategy for septic shock-related ALI.
Area of Science:
- Molecular Biology
- Immunology
- Pathology
Background:
- Nuclear factor-kappa B (NF-κB) pathway activation is implicated in lipopolysaccharide (LPS)-induced acute lung injury (ALI) and inflammation.
- Inhibition of the NF-κB pathway holds potential therapeutic value for ALI.
- MicroRNAs (miRNAs) play critical roles in regulating inflammatory responses and disease pathogenesis.
Purpose of the Study:
- To investigate the role of miR-326 in LPS-induced septic shock with ALI in mice.
- To elucidate the underlying mechanism involving the NF-κB signaling pathway and its potential target, BCL2A1.
- To assess the therapeutic potential of modulating miR-326 in ALI.
Main Methods:
- Establishment of LPS-induced septic shock with ALI mouse models.
- Transfection with miR-326 mimic, miR-326 inhibitor, si-BCL2A1, and combined treatments.
- Assessment of physiological parameters (MAP, AP, HR, W/D ratio).
- Measurement of serum inflammatory cytokines (IL-6, IL-10, IL-1β, TNF-α) via ELISA.
- Analysis of gene and protein expression (miR-326, BCL2A1, NF-κB pathway components, iNOS) using RT-qPCR and Western blot.
- Evaluation of cell viability and apoptosis using CCK-8 assay and flow cytometry.
Main Results:
- Upregulated miR-326 and silenced BCL2A1 aggravated ALI, indicated by altered physiological parameters and increased inflammatory cytokine levels.
- Increased expression of inflammatory mediators (IL-6, IL-1β, TNF-α, p-NF-κB p65, iNOS) and decreased BCL2A1 expression were observed with upregulated miR-326.
- Downregulation of miR-326 reversed these pathological changes, suggesting a protective role.
- MiR-326 directly targets BCL2A1, leading to NF-κB pathway activation and exacerbation of inflammation and lung injury.
Conclusions:
- MiR-326 targeting of BCL2A1 activates the NF-κB signaling pathway, worsening inflammatory responses and lung injury in septic shock with ALI.
- Modulating miR-326 expression presents a potential therapeutic avenue for managing ALI associated with septic shock.
- Further research into miR-326 as a therapeutic target is warranted for ALI treatment.
Related Concept Videos
02:26NF-κB-dependent Signaling Pathway
NF-κB-dependent Signaling Mechanism
The...
03:14Notch Signaling Pathway
The Notch gene came into the limelight in 1914 after the discovery that its mutation in Drosophila melanogaster leads to a serrated (or "notched") wing margin phenotype. It was not...
02:33Hedgehog Signaling Pathway
01:19Interactions Between Signaling Pathways
Convergence and divergence, and cross-talk between signaling pathways
Two distinct signaling pathways can converge on a single functional unit, which may either be a single protein or a complex of proteins. The response is either functionally distinct or synergistic between the two pathways but different from the response...
01:22MicroRNAs
01:41Non-Canonical Wnt Signaling Pathways