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Updated: Sep 26, 2025
A Quantitative Detection Method for MicroRNAs in the Kidney of an Ischemic Kidney Injury Mouse Model
Published on: September 11, 2020
Inhibition of microRNA-543 alleviates sepsis-induced acute kidney injury via targeting Bcl-2
1Department of Critical Care Medicine, Yantai Affiliated Hospital of Binzhou Medical University, Yantai, China. vikyz1@163.com.
Objective:
Sepsis has a high morbidity and mortality and is prone to cause acute kidney injury (AKI). Here, we aimed to demonstrate the function and molecular mechanism of microRNA-543 (miR-543) in septic AKI.
Materials And Methods:
MiR-543 inhibitor or NC was transfected into LPS-treated HK-2 cells to observe lipopolysaccharide (LPS)-induced inflammation and apoptosis. The detection of inflammation and apoptosis of HK-2 cells relies on Western blot, quantitative Reverse-Transcription Polymerase Chain Reaction (qRT-PCR), enzyme-linked immunosorbent assay (ELISA), Cell Counting Kit-8 (CCK-8) assay, flow cytometry, and terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end labeling (TUNEL) staining.
Results:
MiR-543 expression was increased in LPS-treated HK-2 cells. By transfecting miR-543 inhibitor into HK-2 cells, miR-543 expression was dramatically reduced. The downregulation of miR-543 remarkably inhibited the inflammation and apoptosis, which was manifested by the reduction of inflammatory cytokines (TNF-α, IL-6, IL-1β), the reversal of apoptosis-related proteins expression (Bcl-1, Bax), the increase of cell viability and the decrease of the proportion of apoptotic cells. The result of Luciferase activity assay demonstrated that miR-543 directly targets Bcl-2.
Conclusions:
MiR-543 expression was increased in LPS-treated HK-2 cells, and silencing miR-543 could inhibit LPS-induced inflammation and apoptosis in HK-2 cells via targeting Bcl-2.
Insights
MicroRNA-543 (miR-543) is upregulated in sepsis-induced acute kidney injury (AKI). Silencing miR-543 reduces inflammation and apoptosis in kidney cells by targeting Bcl-2.
Area of Science:
- Nephrology
- Molecular Biology
- Biochemistry
Background:
- Sepsis is a life-threatening condition associated with high morbidity and mortality.
- Sepsis frequently leads to acute kidney injury (AKI), a critical complication.
- Understanding the molecular mechanisms underlying septic AKI is crucial for developing effective treatments.
Purpose of the Study:
- To investigate the role and molecular mechanism of microRNA-543 (miR-543) in sepsis-induced acute kidney injury (AKI).
- To elucidate the functional impact of miR-543 on kidney cells exposed to lipopolysaccharide (LPS).
Main Methods:
- Lipopolysaccharide (LPS)-induced HK-2 cell model of septic AKI.
- Transfection with miR-543 inhibitors or negative control (NC).
- Assessment of inflammation and apoptosis using Western blot, qRT-PCR, ELISA, CCK-8 assay, flow cytometry, and TUNEL staining.
- Luciferase activity assay to determine direct targeting.
Main Results:
- miR-543 expression was significantly increased in LPS-treated HK-2 cells.
- Downregulation of miR-543 markedly inhibited LPS-induced inflammation and apoptosis.
- Inhibition of miR-543 reduced pro-inflammatory cytokines (TNF-α, IL-6, IL-1β) and apoptosis markers (Bax), while increasing cell viability and Bcl-2 expression.
- miR-543 was confirmed to directly target Bcl-2.
Conclusions:
- miR-543 expression is elevated in septic AKI.
- Silencing miR-543 mitigates LPS-induced inflammation and apoptosis in kidney cells.
- The protective effect of miR-543 inhibition is mediated through targeting Bcl-2, highlighting a novel therapeutic target for septic AKI.
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