open access

Vol 76, No 6 (2018)
Original articles
Published online: 2018-01-26
Submitted: 2017-08-17
Accepted: 2018-01-26
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MiR-130a inhibition protects rat cardiac myocytes from hypoxia-triggered apoptosis by targeting Smad4

Yuanshi Li, Yingrong Du, Junxian Cao, Qianping Gao, Hongjuan Li, Yangjun Chen, Nihong Lu
DOI: 10.5603/KP.a2018.0040
·
Kardiol Pol 2018;76(6):993-1001.

open access

Vol 76, No 6 (2018)
Original articles
Published online: 2018-01-26
Submitted: 2017-08-17
Accepted: 2018-01-26

Abstract

Background: Cardiomyocyte death facilitates the pathological process underlying ischaemic heart diseases, such as myocardial infarction. Emerging evidence suggests that microRNAs play a critical role in the pathological process underlying myocardial infarction by regulating cardiomyocyte apoptosis. However, the relevance of miR-130a in regulating cardiomyocyte apoptosis and the underlying mechanism are still uncertain.

Aim: We sought to explore the regulatory effect of miR-130a on hypoxic cardiomyocyte apoptosis.

Methods: The expression of miR-130a was measured by quantitative reverse transcription polymerase chain reaction (qRT-PCR). Cell survival was determined by the MTT assay. The lactate dehydrogenase (LDH) assay was performed to deter­mine the severity of hypoxia-induced cell injury. Apoptosis was assessed via caspase-3 analysis. Protein expression level was determined by Western blotting. The genes targeted by miR-130a were predicted using bioinformatics and were validated via the dual-luciferase reporter assay system.

Results: We found that miR-130a expression was greatly increased in hypoxic cardiac myocytes, and that the downregulation of miR-130a effectively shielded cardiac myocytes from hypoxia-triggered apoptosis. In bioinformatic analysis the Smad4 gene was predicted to be the target of miR-130a. This finding was validated through the Western blot assay, dual-luciferase reporter gene assay, and qRT-PCR. MiR-130a inhibition significantly promoted the activation of Smad4 in hypoxic cardiomyocytes. Inter­estingly, knockdown of Smad4 markedly reversed the protective effects induced by miR-130a inhibition. Moreover, we found that the inhibition of miR-130a promoted the activation of transforming growth factor-b1 signalling. Blocking of Smad4 signal­ling significantly abrogated the protective effects of miR-130a inhibition.

Conclusions: The findings indicate that inhibition of miR-130a, which targets the Smad4 gene, shields cardiac myocytes from hypoxic apoptosis. This study offers a novel perspective on the molecular basis of hypoxia-induced cardiomyocyte apoptosis and suggests a possible drug target for the treatment of myocardial infarction.

Abstract

Background: Cardiomyocyte death facilitates the pathological process underlying ischaemic heart diseases, such as myocardial infarction. Emerging evidence suggests that microRNAs play a critical role in the pathological process underlying myocardial infarction by regulating cardiomyocyte apoptosis. However, the relevance of miR-130a in regulating cardiomyocyte apoptosis and the underlying mechanism are still uncertain.

Aim: We sought to explore the regulatory effect of miR-130a on hypoxic cardiomyocyte apoptosis.

Methods: The expression of miR-130a was measured by quantitative reverse transcription polymerase chain reaction (qRT-PCR). Cell survival was determined by the MTT assay. The lactate dehydrogenase (LDH) assay was performed to deter­mine the severity of hypoxia-induced cell injury. Apoptosis was assessed via caspase-3 analysis. Protein expression level was determined by Western blotting. The genes targeted by miR-130a were predicted using bioinformatics and were validated via the dual-luciferase reporter assay system.

Results: We found that miR-130a expression was greatly increased in hypoxic cardiac myocytes, and that the downregulation of miR-130a effectively shielded cardiac myocytes from hypoxia-triggered apoptosis. In bioinformatic analysis the Smad4 gene was predicted to be the target of miR-130a. This finding was validated through the Western blot assay, dual-luciferase reporter gene assay, and qRT-PCR. MiR-130a inhibition significantly promoted the activation of Smad4 in hypoxic cardiomyocytes. Inter­estingly, knockdown of Smad4 markedly reversed the protective effects induced by miR-130a inhibition. Moreover, we found that the inhibition of miR-130a promoted the activation of transforming growth factor-b1 signalling. Blocking of Smad4 signal­ling significantly abrogated the protective effects of miR-130a inhibition.

Conclusions: The findings indicate that inhibition of miR-130a, which targets the Smad4 gene, shields cardiac myocytes from hypoxic apoptosis. This study offers a novel perspective on the molecular basis of hypoxia-induced cardiomyocyte apoptosis and suggests a possible drug target for the treatment of myocardial infarction.

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Keywords

apoptosis, cardiomyocyte, hypoxia, miR-130a, Smad4

About this article
Title

MiR-130a inhibition protects rat cardiac myocytes from hypoxia-triggered apoptosis by targeting Smad4

Journal

Kardiologia Polska (Polish Heart Journal)

Issue

Vol 76, No 6 (2018)

Pages

993-1001

Published online

2018-01-26

DOI

10.5603/KP.a2018.0040

Bibliographic record

Kardiol Pol 2018;76(6):993-1001.

Keywords

apoptosis
cardiomyocyte
hypoxia
miR-130a
Smad4

Authors

Yuanshi Li
Yingrong Du
Junxian Cao
Qianping Gao
Hongjuan Li
Yangjun Chen
Nihong Lu

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