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14 July 2016 : Animal Research  

Inhibition of MiR-92a May Protect Endothelial Cells After Acute Myocardial Infarction in Rats: Role of KLF2/4

Hongxia LiuBCDE, Guofen LiBD, Wenxue ZhaoAF, Yibo HuBCDE

DOI: 10.12659/MSM.897266

Med Sci Monit 2016; 22:2451-2462

Abstract

BACKGROUND: This study was designed to investigate the effects of microRNA-92 (miR-92), Kruppel-like factor 2 (KLF2), and Kruppel-like factor 4 (KLF4) on endothelial injury after acute myocardial infarction (AMI).

MATERIAL AND METHODS: Blood samples were collected from 50 AMI patients for detection of cardiac troponin I (cTnI), heart-type fatty acid-binding protein (H-FABP), and von Willebrand factor (vWF). The Sprague-Dawley rat models of AMI (n=30) were established by ligating their left anterior descending coronary artery. The cardiac markers of AMI patients and rat models were analyzed with enzyme-linked immunosorbent assay and immunohistochemistry. Human umbilical vein endothelial cells were processed into 5 groups: control, negative control, miR-92a inhibitors, miR-92a inhibitors + KLF2 small interfering RNA (siRNA), and miR-92a inhibitors + KLF4 siRNA. Cell proliferation and apoptosis were detected using MTT assay and flow cytometry. RT-PCR and Western blot were conducted to analyze KLF2 and KLF4 expressions.

RESULTS: AMI patients exhibited significantly higher expression of both endothelial injury markers (e.g., cTnI, H-FABP, vWF) and miR-92a in blood samples, when compared with controls (P<0.05). Model rats also had similar expressional tendencies, along with lower KLF2 and KLF4 expressions (P<0.05). Further, it could be observed in cellular experiments that treatment of miR-92a mimics can further upregulate endothelial injury markers, and miR-92a and both KLF2 and KLF4 were downregulated by miR-92a mimics (all, P<0.05). Also, the luciferase activity assay confirmed the direct binding of miR-92a to 3’ UTR of KLF2/4.

CONCLUSIONS: MiR-92a was involved in the endothelial injury process after AMI and was able to suppress KLF2 and KLF4 expression.

Keywords: Aged, 80 and over, Endothelial Cells - metabolism, Fatty Acid-Binding Proteins - metabolism, Human Umbilical Vein Endothelial Cells, Kruppel-Like Transcription Factors - metabolism, MicroRNAs - genetics, Myocardial Infarction - genetics, Rats, Rats, Sprague-Dawley, Signal Transduction, Troponin I - metabolism, von Willebrand Factor - metabolism

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Medical Science Monitor eISSN: 1643-3750
Medical Science Monitor eISSN: 1643-3750