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Updated: Sep 1, 2025
Bronchoalveolar Lavage Exosomes in Lipopolysaccharide-induced Septic Lung Injury
Published on: May 21, 2018
MiR-135a is highly expressed and aggravates inflammatory response in sepsis by targeting MYOM1
Sheng Chen1, Han Zhang2, Haoming Li2
11Department of Cardiovascular Surgery, Fujian Medical University, Union Hospital, Fuzhou, 350001, China; 2Key Laboratory of Cardio-Thoracic Surgery, Fujian Medical University, Fujian Province University, Fuzhou, 350001, China.
Background:
Our research attempted to explore the effect of miR-135a on lipopolysaccharide (LPS)-induced THP-1cells damage and its potential mechanism.
Methods:
LPS (1 μg/mL) was selected to mimic the injury of sepsis in vitro. qRT-PCR was used to detect miR-135a expression. The association between miR-135a and Myomesin 1 (MYOM1) was speculated bythe predication website and confirmed by the dual-luciferase assay. MYOM1 expression was observed by qRT-PCR and western blotting assays. The biological properties of THP-1cells were analyzed by cell counting kit-8 and flow cytometry assays. The concentration of TNF-α, IL-6 and IL-8 in cell supernatant was calculated by enzyme-linked immunosorbent assay.
Results:
A marked augmentation of miR-135a was observed in LPS-induced THP-1 cells. Moreover, depletion of miR-135a alleviated the LPS-induced THP-1 cells injury from the perspective of improving cell viability and reducing cell apoptosis. Importantly, MYOM1, which was under expressed in LPS-induced THP-1 cells, was identified as a target of miR-135a and negatively regulated by miR-135a. Additionally, the mitigative impact of miR-135a inhibitor on THP-1cells damage triggered by LPS was suppressed by MYOM1 depletion.
Conclusions:
Our study suggested that the protective effect of miR-135a inhibitor on LPS-induced THP-1 cells injury was realized by regulation of MYOM1, which might afford a pair of novel molecules for sepsis clinical diagnosis.
Insights
MicroRNA-135a (miR-135a) protects against lipopolysaccharide (LPS)-induced cell damage by regulating Myomesin 1 (MYOM1). This finding offers potential new biomarkers for sepsis diagnosis.
Area of Science:
- Molecular Biology
- Cell Biology
- Biochemistry
Background:
- Lipopolysaccharide (LPS) induces damage in THP-1 cells, mimicking sepsis.
- Investigating the role of microRNA-135a (miR-135a) in this cellular injury model.
Purpose of the Study:
- To explore the effect of miR-135a on LPS-induced THP-1 cell damage.
- To elucidate the underlying molecular mechanism involving Myomesin 1 (MYOM1).
Main Methods:
- LPS stimulation of THP-1 cells in vitro.
- Quantitative real-time PCR (qRT-PCR) for gene expression analysis.
- Dual-luciferase assay to confirm miR-135a and MYOM1 interaction.
- Cell viability (CCK-8) and apoptosis assays (flow cytometry).
- Enzyme-linked immunosorbent assay (ELISA) for cytokine levels (TNF-α, IL-6, IL-8).
Main Results:
- miR-135a expression was significantly upregulated in LPS-treated THP-1 cells.
- Downregulation of miR-135a mitigated LPS-induced cell injury, improving viability and reducing apoptosis.
- MYOM1 was identified as a direct target of miR-135a, with its expression being downregulated by LPS.
- MYOM1 depletion reversed the protective effects of miR-135a inhibition on LPS-induced cell damage.
Conclusions:
- miR-135a plays a protective role against LPS-induced THP-1 cell injury.
- The protective mechanism involves the negative regulation of MYOM1 by miR-135a.
- miR-135a and MYOM1 represent potential novel molecular targets for sepsis diagnosis and treatment.
