MicroRNA‑29a/b/c targets iNOS and is involved in protective remote ischemic preconditioning in an ischemia‑reperfusion rat model of non‑alcoholic fatty liver disease

  • Authors:
    • Yun‑Fei Duan
    • Dong‑Lin Sun
    • Jing Chen
    • Feng Zhu
    • Yong An
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  • Published online on: January 18, 2017     https://doi.org/10.3892/ol.2017.5623
  • Pages: 1775-1782
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Abstract

Remote ischemic preconditioning (RIPC) protects against the injury that is incurred by ischemia and reperfusion (IR); however, the role of RIPC in liver IR injury in non‑alcoholic fatty liver disease (NAFLD) remains unclear. In this study, a NAFLD rat model was utilized in a series of different surgical procedures and molecular experiments. Rats of the IR group and the RIPC+IR group exhibited more severe injury than NAFLD control rats (in which the liver was prodded following a median‑incision laparotomy). The liver condition, measured by serum alanine transaminase and aspartate transaminase levels, of the RIPC+IR group was better than that of the IR group. In addition, alanine transaminase and aspartate transaminase levels were lower in the RIPC+IR group compared with the IR group (P<0.001). Flow cytometry revealed that the cell apoptosis ratio was significantly lower in the RIPC+IR group than in the IR group (P<0.001). Reverse transcription‑polymerase chain reaction (RT‑qPCR) was used to assess miR‑29a/b/c levels, revealing that they were significantly reduced in the RIPC and RIPC+IR groups, but did not vary in the IR group compared with the control group. RT‑qPCR also revealed that iNOS mRNA levels were not significantly different among any of the NAFLD groups; however, western blot analysis indicated that iNOS protein levels were increased in the RIPC group and the RIPC+IR group compared with the control and IR groups. A luciferase reporter assay demonstrated that transfection with miR‑29a/b/c mimics significantly decreased the luciferase activities of plasmids containing the wild‑type iNOS 3'‑untranslated region (UTR) (relative fluorescence intensity: 0.47±0.06 for miR‑29a, 0.36±0.07 for miR‑29b, 0.41±0.04 for miR‑29c; P<0.001), whereas the activities of plasmids containing the mutant iNOS 3'‑UTR sequence were not markedly affected [relative fluorescence intensity: 0.99±0.08 for miR‑29a (P=0.1349), 0.99±0.09 for miR‑29b (P=0.1607), 0.97±0.07 for miR‑29c (P=0.1824)]. This suggested that miR‑29a/b/c downregulates iNOS by directly targeting its 3'‑UTR. In summary, the results suggest that RIPC has a protective effect in NAFLD liver IR injury, which may be due to reduced miR‑29a/b/c levels in the skeletal muscle, leading to increased iNOS and, therefore, nitric oxide.
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March-2017
Volume 13 Issue 3

Print ISSN: 1792-1074
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Spandidos Publications style
Duan YF, Sun DL, Chen J, Zhu F and An Y: MicroRNA‑29a/b/c targets iNOS and is involved in protective remote ischemic preconditioning in an ischemia‑reperfusion rat model of non‑alcoholic fatty liver disease. Oncol Lett 13: 1775-1782, 2017
APA
Duan, Y., Sun, D., Chen, J., Zhu, F., & An, Y. (2017). MicroRNA‑29a/b/c targets iNOS and is involved in protective remote ischemic preconditioning in an ischemia‑reperfusion rat model of non‑alcoholic fatty liver disease. Oncology Letters, 13, 1775-1782. https://doi.org/10.3892/ol.2017.5623
MLA
Duan, Y., Sun, D., Chen, J., Zhu, F., An, Y."MicroRNA‑29a/b/c targets iNOS and is involved in protective remote ischemic preconditioning in an ischemia‑reperfusion rat model of non‑alcoholic fatty liver disease". Oncology Letters 13.3 (2017): 1775-1782.
Chicago
Duan, Y., Sun, D., Chen, J., Zhu, F., An, Y."MicroRNA‑29a/b/c targets iNOS and is involved in protective remote ischemic preconditioning in an ischemia‑reperfusion rat model of non‑alcoholic fatty liver disease". Oncology Letters 13, no. 3 (2017): 1775-1782. https://doi.org/10.3892/ol.2017.5623