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Insulin ameliorates miR-1-induced injury in H9c2 cells under oxidative stress via Akt activation

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Abstract

Growing evidence indicates that aberrant upregulation of microRNA-1 (miR-1) occurs in ischemic myocardium. In addition, insulin elicits metabolism-independent cardioprotection against cardiovascular diseases. The aim of this study is to determine whether insulin ameliorates miR-1-induced injury in H9c2 cells under oxidative stress and to investigate the underlying mechanisms. By quantitative real-time RT-PCR (qRT-PCR), we show that miR-1 is upregulated in H9c2 cells after treatment with hydrogen peroxide (H2O2), and this effect is both dose- and time dependent. Furthermore, expression of miR-1 decreased significantly after insulin treatment (4.5 ± 0.1 vs. 3.0 ± 0.2, p < 0.05). To determine the potential role of miR-1 in cellular injury and gene regulation, adenovirus-mediated overexpression of miR-1 was used. Overexpression of miR-1 decreased cell viability by 28 ± 2 % (n = 6, p < 0.05) and damaged Akt activation with or without H2O2 treatment. To further investigate the effect of the phosphatidylinositol 3-kinase (PI3K)/Akt pathway in miR-1-induced injury, H9c2 cells were pretreated with LY294002 (10 μM LY, a specific inhibitor of PI3K) with or without insulin (100 nM) and subjected to H2O2 treatment. LY pretreatment-inhibited Akt activation, lead to increased reactive oxygen species (ROS), and further decreased cell viability induced by miR-1 (n = 6, p < 0.05, n = 9–10 cells/group, p < 0.05 and n = 6, p < 0.05) under oxidative stress. This effect was abolished by insulin. In summary, our findings suggest that miR-1expression is sensitive to H2O2 stimulation. In addition, insulin decreases miR-1 expression and induces a marked protective effect on miR-1-induced injury under oxidative stress, which may be mediated by the Akt-mediated pathway. These results provide an important, novel clue as to the mechanism of the cardiovascular action of insulin.

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Acknowledgments

This work was partly supported by the National Basic Research Program of China (973 Program: No. 2012CB51780X), National Science Fund for Distinguished Young Scholars (81025002 to Z.Y.), and Natural Science Foundation of China (30871043 to Z.Y.).

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Authors and Affiliations

  1. Department of Cardiovascular Medicine, First Affiliated Hospital of Medical School, Xi’an Jiaotong University, 277 Yanta West Road, Xi’an, 710061, Shaanxi, China

    Tao Chen & Zuyi Yuan

  2. Department of Pediatrics, Emory+Children’s Center for Cardiovascular Biology, Children’s Healthcare of Atlanta, Emory University School of Medicine, Atlanta, GA, 30322, USA

    Tao Chen, Guoliang Ding & Mary B. Wagner

  3. Key Laboratory of Environment and Genes Related to Diseases, Xi’an Jiaotong University, Ministry of Education, Xi’an, 710061, Shaanxi, China

    Zuyi Yuan

  4. School of Basic Medicine, Zhejiang Chinese Medical University, Hangzhou, 310053, Zhejiang, China

    Zhuqing Jin

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  1. Tao Chen
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  2. Guoliang Ding
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  3. Zhuqing Jin
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  4. Mary B. Wagner
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  5. Zuyi Yuan
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Correspondence to Mary B. Wagner or Zuyi Yuan.

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Chen, T., Ding, G., Jin, Z. et al. Insulin ameliorates miR-1-induced injury in H9c2 cells under oxidative stress via Akt activation. Mol Cell Biochem 369, 167–174 (2012). https://doi.org/10.1007/s11010-012-1379-7

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  • DOI: https://doi.org/10.1007/s11010-012-1379-7

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