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MiR-539-5p inhibits the inflammatory injury in septic H9c2 cells by regulating IRAK3
1Department of Emergency, Children's Hospital of Nanjing Medical University, No. 72 Guangzhou Road, Gulou District, Nanjing, Jiangsu, 210008, China.
Background:
MicroRNAs (miRNAs) have been confirmed to play a potential role in sepsis, but little is known about their role in sepsis-induced cardiomyopathy (SIC).
Methods:
The model of septic cardiomyopathy was constructed with H9c2 cells induced by lipopolysaccharide (LPS), and the expression of miR-539-5p was detected by qRT-PCR assay. ELISA, CCK-8, EdU TUNEL analysis were performed to evaluate the role of miR-539-5p in inflammation response, viability, proliferation and apoptosis of LPS-treated H9c2 cells. Moreover, miRWalk and TargetScan prediction, and dual-luciferase reporter gene assays were carried out to predict and confirm the target of miR-539-5p. Furthermore, the effects of target on inflammation response, proliferation and apoptosis of LPS-induced H9c2 cells mediated by miR-539-5p was further explored.
Results:
The expression of miR-539-5p was obviously down-regulated in LPS-induced H9c2 cells. In addition, over-expression of miR-539-5p significantly inhibited the inflammation response, promoted viability and proliferation, and suppressed apoptosis of LPS-treated H9c2 cells. Moreover, interleukin-1 receptor-associated kinase 3 (IRAK3) was verified as a target of miR-539-5p by dual-luciferase reporter gene assay. Besides, IRAK3 was highly expressed in H9c2 cells transfected with miR-539-5p inhibitor detected with qRT-PCR and western blot assays. Furthermore, over-expression of IRAK3 partially weakened the effects of miR-539-5p mimic on the inflammation response, proliferation and apoptosis of LPS-induced H9c2 cells.
Conclusions:
MiR-539-5p potentially plays an important role in the pathogenesis of LPS-induced sepsis by targeting IRAK3, suggesting that miR-539-5p may be a potential new target for the treatment of LPS-induced sepsis.
Insights
MicroRNA-539-5p is downregulated in sepsis-induced cardiomyopathy. Upregulating miR-539-5p protects against inflammation and cell death by targeting IRAK3, suggesting it as a potential therapeutic target.
Area of Science:
- Cardiology
- Molecular Biology
- Genetics
Background:
- Sepsis-induced cardiomyopathy (SIC) is a serious complication of sepsis.
- The role of microRNAs (miRNAs) in SIC is not well understood.
- This study investigates the specific role of miR-539-5p in LPS-induced cardiomyopathy.
Purpose of the Study:
- To investigate the role of miR-539-5p in lipopolysaccharide (LPS)-induced H9c2 cell injury.
- To identify the target gene of miR-539-5p and explore its mechanism in LPS-induced cardiomyopathy.
- To evaluate miR-539-5p as a potential therapeutic target for SIC.
Main Methods:
- Constructed a model of septic cardiomyopathy using LPS-treated H9c2 cells.
- Quantified miR-539-5p expression using qRT-PCR.
- Assessed cell viability, proliferation, and apoptosis via CCK-8, EdU, and TUNEL assays.
- Utilized dual-luciferase reporter assays to confirm IRAK3 as a direct target of miR-539-5p.
Main Results:
- miR-539-5p expression was significantly downregulated in LPS-induced H9c2 cells.
- Overexpression of miR-539-5p inhibited inflammation, promoted cell viability and proliferation, and suppressed apoptosis.
- Interleukin-1 receptor-associated kinase 3 (IRAK3) was confirmed as a direct target of miR-539-5p.
- IRAK3 overexpression partially reversed the protective effects of miR-539-5p.
Conclusions:
- miR-539-5p plays a protective role in LPS-induced cardiomyopathy by targeting IRAK3.
- miR-539-5p may serve as a novel therapeutic target for sepsis-induced cardiomyopathy.
