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Updated: Jun 28, 2025
Bronchoalveolar Lavage Exosomes in Lipopolysaccharide-induced Septic Lung Injury
Published on: May 21, 2018
Macrophage-derived exosomal miRNA-141 triggers endothelial cell pyroptosis by targeting NLRP3 to accelerate sepsis
Feng Zhan1, Jun Zhang1, Ping He1
1Department of Emergency Medicine, Hainan General Hospital, Hainan Affiliated Hospital of Hainan Medical University, Haikou, China.
Abstract:
Sepsis, critical condition marked by severe organ dysfunction from uncontrolled infection, involves the endothelium significantly. Macrophages, through paracrine actions, play a vital role in sepsis, but their mechanisms in sepsis pathogenesis remain elusive. Objective: We aimed to explore how macrophage-derived exosomes with low miR-141 expression promote pyroptosis in endothelial cells (ECs). Exosomes from THP-1 cell supernatant were isolated and characterized. The effects of miR-141 mimic/inhibitor on apoptosis, proliferation, and invasion of Human Umbilical Vein Endothelial Cells (HUVECs) were assessed using flow cytometry, CCK-8, and transwell assays. Key pyroptosis-related proteins, including caspase-1, IL-18, IL-1β, NLR Family Pyrin Domain Containing 3 (NLRP3), ASC, and cleaved-GSDMD, were analyzed via Western blot. The interaction between miR-141 and NLRP3 was studied using RNAhybrid v2.2 and dual-Luciferase reporter assays. The mRNA and protein level of NLRP3 after exosomal miR-141 inhibitor treatment was detected by qPCR and Western blot, respectively. Exosomes were successfully isolated. miR-141 mimic reduced cell death and pyroptosis-related protein expression in HUVECs, while the inhibitor had opposite effects, increasing cell death, and enhancing pyroptosis protein expression. Additionally, macrophage-derived exosomal miR-141 inhibitor increased cell death and pyroptosis-related proteins in HUVECs. miR-141 inhibits NLRP3 transcription. Macrophages facilitate sepsis progression by secreting miR-141 decreased exosomes to activate NLRP3-mediated pyroptosis in ECs, which could be a potentially valuable target of sepsis therapy.
Insights
Macrophages release exosomes low in miR-141, promoting pyroptosis in endothelial cells and worsening sepsis. Inhibiting this process offers a potential sepsis therapy target.
Area of Science:
- Cell Biology
- Immunology
- Pathophysiology
Background:
- Sepsis involves significant endothelial dysfunction and macrophage paracrine actions.
- Mechanisms of macrophage involvement in sepsis pathogenesis are not fully understood.
- Endothelial cell pyroptosis is implicated in sepsis progression.
Purpose of the Study:
- To investigate how macrophage-derived exosomes with low miR-141 expression induce pyroptosis in endothelial cells (ECs).
- To explore the therapeutic potential of targeting the miR-141/NLRP3 axis in sepsis.
Main Methods:
- Isolation and characterization of exosomes from THP-1 cells.
- Assessment of miR-141 mimic/inhibitor effects on Human Umbilical Vein Endothelial Cells (HUVECs) using flow cytometry, CCK-8, and transwell assays.
- Analysis of pyroptosis-related proteins (caspase-1, IL-18, IL-1β, NLRP3, ASC, cleaved-GSDMD) via Western blot and qPCR.
- Investigation of miR-141 and NLRP3 interaction using RNAhybrid and dual-luciferase reporter assays.
Main Results:
- Macrophage-derived exosomes were successfully isolated.
- miR-141 mimic reduced HUVEC pyroptosis, while miR-141 inhibitor increased it.
- Exosomal miR-141 inhibitor treatment enhanced pyroptosis in HUVECs.
- miR-141 was found to directly inhibit NLRP3 transcription.
Conclusions:
- Macrophages contribute to sepsis by secreting exosomes with reduced miR-141, activating NLRP3-mediated pyroptosis in endothelial cells.
- Targeting the NLRP3 inflammasome or modulating exosomal miR-141 levels presents a promising therapeutic strategy for sepsis.
