Angiotensin II upregulated the expression of microRNA-224 but not microRNA-21 in adult rat cardiac fibroblasts.

The role of microRNA-21 (miRNA-21, miR-21) in cardiac fibrosis remains controversial, while the role of microRNA-224 (miRNA-224, miR-224) in cardiac fibroblasts has not been reported. Angiotensin II (Ang II) is known to play a pivotal role in the pathogenesis of cardiac fibrosis. The aim of this study was to confirm whether the expression of miR-21 and miR-224 is regulated by Ang II in adult rat cardiac fibroblasts. Semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) and quantitative PCR (qPCR) were performed to measure the levels of miR-21 and miR-224 in Ang II-treated or untreated adult rat cardiac fibroblasts. The RT-PCR, qPCR and previous miRNA array results demonstrated that treatment with Ang II (100 nM) for 24 h did not induce the increase of miR-21 in cardiac fibroblasts, although the level of miR-21 in cardiac fibroblasts was not considered as low. The results of the present study also demonstrated that Ang II significantly upregulated the expression of miR-224 in adult rat cardiac fibroblasts. Bioinformatic analysis revealed that the potential target genes of miR-224 included SMAD4, SMAD5, cyclin-dependent kinase 9 and early growth response 1/2. In previous studies, it was reported that miR-224 was upregulated in tumors by promoting cell proliferation and targeting SMAD4. Those results indicated the potential roles of miR-224 in cardiac fibroblasts and cardiac fibrosis. In conclusion, results of the present study demonstrated that miR-21 was not induced by Ang II, whereas Ang II upregulated miR-224 expression in adult rat cardiac fibroblasts, a finding that may provide a starting point for the investigation of the potential role of miR-224 in cardiac fibrosis.