Abstract
This study aims to investigate the role of targeting lncRNA myocardial infarction-associated transcript (MIAT) in protection against hypoxia/reoxygenation (H/R) injury in H9c2 cells in vitro and myocardial ischemia/reperfusion (I/R) injury in vivo by regulating expression of NF-kB and p53 upregulated modulator of apoptosis (PUMA). H9C2 cells were infected with lentivirus expressing the short-hairpin RNA direct against human MIAT gene (Lv-MIAT shRNA) or lentivirus expressing scrambled control (Lv-NC shRNA) or PUMA siRNA or p65 siRNA or their control siRNA respectively. Then the H9c2 cells were infected with Lv-shRNA to 2 hours of hypoxia (H) and 24 hour of reoxygenation (R). 100 ul of Lv-MIAT shRNA (1 × 108 PFU) or Lv-NC shRNA was transfected into mouse hearts, then the hearts were subjected to I/R (1h/72 h). We discovered targeting MIAT remarkably enhanced H9c2 cell viability, decreased H/R-induced cell apoptosis and LDH leakage and significantly decreased I/R-induced myocardial infarct size, reduced myocardial apoptosis and enhanced the heart function. Targeting MIAT downregulated p65 nuclear translocation, NF-κB activity and anti-apoptotic protein cleaved-caspase-3, Bax, and upregulated anti-apoptotic protein Bcl-2 induced by H/R or I/R. Our study suggests that targeting MIAT may protect against H9c2 cardiomyoblasts H/R injury or myocardial I/R injury via inhibition of cell apoptosis, mediated by NF-κB and PUMA signal pathway.
Keywords:
Hypoxia/reoxygenation; apoptosis; ischaemia-reperfusion; lncRNA myocardial infarction-associated transcript; nuclear factor kappa B; p53 upregulated modulator of apoptosis.
MeSH terms
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Animals
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Apoptosis / genetics
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Apoptosis Regulatory Proteins / antagonists & inhibitors
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Apoptosis Regulatory Proteins / genetics*
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Apoptosis Regulatory Proteins / metabolism
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Caspase 3 / genetics
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Caspase 3 / metabolism
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Cell Hypoxia
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Cell Line
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Cell Survival
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Gene Expression Regulation
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Humans
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L-Lactate Dehydrogenase / metabolism
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Mice
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Myocardial Reperfusion Injury / genetics*
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Myocardial Reperfusion Injury / metabolism
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Myocardial Reperfusion Injury / pathology
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Myocardial Reperfusion Injury / prevention & control
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Myocardium / metabolism*
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Myocardium / pathology
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Myocytes, Cardiac / metabolism*
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Myocytes, Cardiac / pathology
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Protein Transport
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Proto-Oncogene Proteins c-bcl-2 / genetics
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Proto-Oncogene Proteins c-bcl-2 / metabolism
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RNA, Long Noncoding / antagonists & inhibitors
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RNA, Long Noncoding / genetics*
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RNA, Long Noncoding / metabolism
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RNA, Small Interfering / genetics
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RNA, Small Interfering / metabolism
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Rats
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Signal Transduction
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Transcription Factor RelA / antagonists & inhibitors
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Transcription Factor RelA / genetics*
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Transcription Factor RelA / metabolism
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Tumor Suppressor Proteins / antagonists & inhibitors
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Tumor Suppressor Proteins / genetics*
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Tumor Suppressor Proteins / metabolism
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bcl-2-Associated X Protein / genetics
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bcl-2-Associated X Protein / metabolism
Substances
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Apoptosis Regulatory Proteins
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Bax protein, mouse
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Miat long non-coding RNA
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PUMA protein, mouse
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Proto-Oncogene Proteins c-bcl-2
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RNA, Long Noncoding
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RNA, Small Interfering
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Rela protein, mouse
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Transcription Factor RelA
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Tumor Suppressor Proteins
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bcl-2-Associated X Protein
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Bcl2 protein, mouse
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L-Lactate Dehydrogenase
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Casp3 protein, mouse
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Caspase 3