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Resveratrol protects H9c2 cells against hypoxia-induced apoptosis through miR-30d-5p/SIRT1/NF-κB axis

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Abstract

Previous studies have demonstrated the cardioprotective role of resveratrol (Res). However, the underlying molecular mechanisms involved in the protective role of Res are still largely unknown. H9c2 cells were distributed into five groups: normal condition (Control), DMSO, 20 mMRes (dissolved with DMSO), hypoxia (Hyp), and Res+Hyp. Cell apoptosis was evaluated using flow cytometry and protein analysis of cleaved caspase 3 (cle-caspase 3). qRT-PCR assay was performed to measure the expression of microRNA-30d-5p (miR-30d-5p). MTT assay was performed to evaluate the cell proliferation. The relationship between miR-30d-5p and silent information regulator 1 (SIRT1) was confirmed by luciferase reporter, RNA immunoprecipitation (RIP), and western blot assays. Western blot was performed to analyze NF-κB/p65 and I-κBα expressions. Our data showed that hypoxia enhanced apoptosis andNF-κB signaling pathway, which was alleviated by Res treatment. Hypoxia increased the expression of miR-30d-5p while decreased the SIRT1expression, which was also attenuated by Res treatment. Furthermore, miR-30d-5p depletion inhibited the proliferation, reduced apoptosis and decreased the expression of cle-caspase 3 in H9c2 cells with hypoxia treatment. Luciferase reporter, RIP, and western blot assays further confirmed that miR-30d-5p negatively regulated the expression of SIRT1. Interestingly, the rescue-of-function experiments further indicated that knockdown of SIRT1 attenuated the effect of miR-30d-5p depletion on proliferation, apoptosis NF-κB signaling pathway inH9c2 cells with hypoxia treatment. In addition, the suppression of NF-κB signaling pathway increased cell viability while decreased cell apoptosis in hypoxia-mediatedH9c2 cells. Our data suggested Res mayprotectH9c2 cells against hypoxia-induced apoptosis through miR-30d-5p/SIRT1/NF-κB axis.

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Author information

Correspondence to Zeng Tang Zhang or Tao Li.

Additional information

Corresponding editor: BJ Rao

Communicated by BJ RAO.

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Han, X., Zhang, L., Liu, Y. et al. Resveratrol protects H9c2 cells against hypoxia-induced apoptosis through miR-30d-5p/SIRT1/NF-κB axis. J Biosci 45, 42 (2020). https://doi.org/10.1007/s12038-020-9997-9

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Keywords

  • Apoptosis
  • hypoxia
  • miR-30d-5p/SIRT1/NF-κB axis
  • resveratrol