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Purification and Transplantation of Myogenic Progenitor Cell Derived Exosomes to Improve Cardiac Function in Duchenne Muscular Dystrophic Mice
Published on: April 10, 2019
Effect of miR-412-5p-loaded exosomes in H9c2 cardiomyocytes via the MAPK pathway
1Department of Anesthesiology and Pain Research Institute, Yonsei University College of Medicine, 50 Yonsei-ro, Seodaemun-gu, Seoul 03722, Republic of Korea.
Objectives:
MicroRNAs (miRNAs) are small non-coding RNAs that function in all biological processes. Recent findings suggest that exosomes, which are small vesicles abundantly secreted by various cell types, can transport miRNAs to target cells. Here, we elucidated the effect of miRNA-loaded exosomes on lipopolysaccharide (LPS)-induced inflammation in H9c2 cardiomyocytes.
Materials And Methods:
Exosomes were isolated from mesenchymal stem cells (MSC) and loaded with miR-412-5p. Additionally, the effect of the miR-412-5p-loaded exosomes on LPS-induced inflammation in H9c2 cardiomyocytes was evaluated by assessing the levels of nitric oxide (NO), reactive oxygen species (ROS), and prostaglandin E2 (PGE2). The expression of cyclooxygenase-2 (COX-2), inducible nitric oxide synthase (iNOS), inflammatory cytokines, and mitogen-activated protein kinase (MAPK) signaling factors was evaluated using reverse transcription-quantitative PCR and western blotting.
Results:
miR-412-5p-loaded exosomes inhibited LPS-induced secretion of inflammatory mediators (NO, PGE2, and ROS), pro-inflammatory cytokines (IL-1β and IL-6), and COX-2 and iNOS expression. Additionally, miR-412-5p-loaded exosomes significantly decreased the expression of MAPK signaling molecules, including p-extracellular signal-regulated kinase (ERK), p-p38, and p-Jun kinase (JNK), in H9c2 cardiomyocytes.
Conclusion:
These findings showed that miR-412-5p-loaded exosomes ameliorated LPS-induced inflammation in H9c2 cardiomyocytes by inhibiting COX-2 and iNOS expression, inflammatory mediators, and pro-inflammatory cytokines via the MAPK pathway. The findings indicate that miR-412-5p-loaded exosomes may be effective for the prevention of myocardial injury.
Insights
Exosomes loaded with miR-412-5p reduce inflammation in heart cells by inhibiting inflammatory mediators and pathways. This suggests potential for preventing heart injury.
Area of Science:
- Cardiovascular Biology
- Molecular Biology
- Cell Biology
Background:
- MicroRNAs (miRNAs) are key regulators of biological processes.
- Exosomes facilitate intercellular communication by transporting miRNAs.
- Lipopolysaccharide (LPS) induces inflammation in H9c2 cardiomyocytes.
Purpose of the Study:
- To investigate the anti-inflammatory effects of miR-412-5p-loaded exosomes in LPS-induced H9c2 cardiomyocyte inflammation.
- To elucidate the underlying molecular mechanisms involving inflammatory mediators and signaling pathways.
Main Methods:
- Exosomes were isolated from mesenchymal stem cells and loaded with miR-412-5p.
- H9c2 cardiomyocytes were treated with LPS and miR-412-5p-loaded exosomes.
- Levels of nitric oxide (NO), reactive oxygen species (ROS), and prostaglandin E2 (PGE2) were measured.
- Expression of COX-2, iNOS, inflammatory cytokines (IL-1β, IL-6), and MAPK signaling factors was assessed via RT-qPCR and Western blotting.
Main Results:
- miR-412-5p-loaded exosomes significantly inhibited LPS-induced NO, PGE2, and ROS secretion.
- Exosomes reduced the expression of COX-2, iNOS, IL-1β, and IL-6.
- Downregulation of MAPK signaling molecules (p-ERK, p-p38, p-JNK) was observed.
Conclusions:
- miR-412-5p-loaded exosomes effectively ameliorate LPS-induced inflammation in H9c2 cardiomyocytes.
- The anti-inflammatory effect is mediated through inhibition of COX-2, iNOS, inflammatory cytokines, and the MAPK pathway.
- These exosomes show promise for preventing myocardial injury.
