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Published on: June 15, 2018
Overexpression of microRNA-133b reduces myocardial injuries in children with viral myocarditis by targeting Rab27B
1Wuhan Children's Hospital (Wuhan Maternal and Child Healthcare Hospital), Tongji Medical College, Huazhong University of Science & Technology, Wuhan 430000, P.R. China.
Abstract:
The present study is to measure the expression of microRNA (miRNA or miR)-133b in circulating blood of children with viral myocarditis before and after drug treatment, and to investigate its relationship with the severity of myocardial lesions. A total of 36 children patients with viral myocarditis who received treatments at our hospital between June 2014 and June 2016 were enrolled in the present study, including 21 boys and 15 girls (age range, 9 months - 16 years).Quantitative real-time polymerase chain reaction was used to determine the expression of miR-133b in peripheral blood of patients and cardiomyocytes infected with CVB3. CCK-8 assay was used to test the proliferation of cardiomyocytes. ELISA was used to determine the levels of creatine kinase (CK-MB) and lactate dehydrogenase (LDH) in peripheral blood and cardiomyocyte culture supernatants. Western blotting and ELISA were performed to measure the levels of tumor necrosis factor-α and interleukin-6 in cardiomyocytes infected by CVB3 and cell culture supernatants. Bioinformatics was used to predict the target gene of miR-133b. Silencing of Rab27B gene was achieved by transfection with its small-interfering RNA. Dual luciferase reporter assay was carried out to test whether miR-133b directly targets Rab27B. Reduced expression of miR-133b in peripheral blood was possibly correlated with myocardial injuries in viral myocarditis miR-133b. Expression of miR-133b was significantly reduced in cardiomyocytes infected with CVB3 virus. Overexpression of miR-133b inhibited cardiomyocyte injuries caused by CVB3 virus infection, and the enhanced production and release of cytokines TNF-α and IL-6 by cardiomyocytes infected with CVB3 virus. Rab27B promoted injuries of cardiomyocytes induced by CVB3 infection and facilitated the synthesis and release of cytokines TNF-α and IL-6 by cardiomyocytes. miR-133b was able to bind to the 3'-untranslated region seeding region of Rab27B mRNA. The present study demonstrates that expression of miR-133b in peripheral blood from children with viral myocarditis is reduced, and negatively correlated with myocardial injuries. miR-133b inhibits the proliferation of cardiomyocytes and the release of cytokines TNF-α and IL-6, and alleviates CVB3 infection-induced myocardial injuries by targeting Rab27B.
Insights
Reduced microRNA-133b (miR-133b) in children with viral myocarditis correlates with heart injury. Restoring miR-133b levels alleviates viral myocarditis by targeting Rab27B, reducing inflammation and cardiomyocyte damage.
Area of Science:
- Cardiology
- Molecular Biology
- Pediatrics
Background:
- Viral myocarditis is a serious condition in children.
- MicroRNAs play crucial roles in cardiovascular diseases.
- Understanding molecular mechanisms is key to effective treatment.
Purpose of the Study:
- To measure miR-133b levels in children with viral myocarditis.
- To investigate the relationship between miR-133b and myocardial injury severity.
- To explore the therapeutic potential of miR-133b in viral myocarditis.
Main Methods:
- Quantitative real-time polymerase chain reaction for miR-133b expression.
- CCK-8 assay for cardiomyocyte proliferation.
- ELISA and Western blotting for inflammatory markers (TNF-α, IL-6) and cardiac enzymes (CK-MB, LDH).
- Bioinformatics and dual luciferase reporter assay to identify and validate miR-133b targets (Rab27B).
Main Results:
- miR-133b expression was significantly reduced in children with viral myocarditis and in CVB3-infected cardiomyocytes.
- Reduced miR-133b levels correlated with myocardial injury severity.
- Overexpression of miR-133b inhibited CVB3-induced cardiomyocyte injury, inflammation (TNF-α, IL-6), and proliferation.
- miR-133b directly targets Rab27B, and Rab27B promotes CVB3-induced cardiomyocyte injury.
Conclusions:
- Circulating miR-133b is a potential biomarker for viral myocarditis severity in children.
- miR-133b exerts protective effects against viral myocarditis by targeting Rab27B.
- Modulating miR-133b levels offers a promising therapeutic strategy for viral myocarditis.
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