Skip to main content

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

Journal of Biosciences volume 45, Article number: 42 (2020) Cite this article

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.

This is a preview of subscription content, access via your institution.

Access options

Buy single article

Instant access to the full article PDF.

USD 39.95

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6

References

  • Abbate A and Narula J 2012 Role of apoptosis in adverse ventricular remodeling. Heart Fail. Clin. 8 79–86

    Article  Google Scholar 

  • Boateng S and Sanborn T 2013 Acute myocardial infarction. Disease-a-Month 59 83–96

    Article  Google Scholar 

  • Bonnefont-Rousselot D 2016 Resveratrol and Cardiovascular Diseases. Nutrients 8 250

    Google Scholar 

  • Boon RA and Dimmeler S 2015 MicroRNAs in myocardial infarction. Nat. Rev. Cardiol. 12 135–142

    CAS  Article  Google Scholar 

  • Chen CJ, Yu W, Fu YC, Wang X, Li JL and Wang W 2009 Resveratrol protects cardiomyocytes from hypoxia-induced apoptosis through the SIRT1-FoxO1 pathway. Biochem. Biophys. Res. Commun. 378 389–393

    CAS  Article  Google Scholar 

  • Chen Y, Zhao Y, Chen W, Xie L, Zhao ZA, Yang J, Chen Y, Lei W et al. 2017 MicroRNA-133 overexpression promotes the therapeutic efficacy of mesenchymal stem cells on acute myocardial infarction. Stem Cell. Res. Ther 8 268

    Article  Google Scholar 

  • Fan Y, Liu L, Fang K, Huang T, Wan L, Liu Y, Zhang S, Yan D et al. 2016 Resveratrol Ameliorates cardiac hypertrophy by down-regulation of mir-155 through activation of breast cancer type 1 susceptibility protein. J. Am. Heart Assoc. 5 e002648

    PubMed  PubMed Central  Google Scholar 

  • Fu BC, Lang JL, Zhang DY, Sun L, Chen W, Liu W, Liu KY, Ma CY et al. 2017 Suppression of miR-34a expression in the myocardium protects against ischemia-reperfusion injury through SIRT1 protective pathway. Stem Cells Dev. 26 1270–1282

    CAS  Article  Google Scholar 

  • Han D, Li X, Li S, Su T, Fan L, Fan WS, Qiao HY, Chen JW et al. 2017 Reduced silent information regulator 1 signaling exacerbates sepsis-induced myocardial injury and mitigates the protective effect of a liver X receptor agonist. Free Radic. Biol. Med. 113 291–303

    CAS  Article  Google Scholar 

  • Hernandez-Jimenez M, Hurtado O, Cuartero MI, Ballesteros I, Moraga A, Pradillo JM, McBurney MW, Lizasoain I et al. 2013 Silent information regulator 1 protects the brain against cerebral ischemic damage. Stroke 44 2333–2337

    CAS  Article  Google Scholar 

  • Hsu CP, Zhai P, Yamamoto T, Maejima Y, Matsushima S, Hariharan N, Shao D, Takagi H et al. 2010 Silent information regulator 1 protects the heart from ischemia/reperfusion. Circulation 122 2170–2182

    Article  Google Scholar 

  • Hwang JT, Kwon DY, Park OJ and Kim MS 2008 Resveratrol protects ROS-induced cell death by activating AMPK in H9c2 cardiac muscle cells. Genes Nutr 2 323–326

    CAS  Article  Google Scholar 

  • Jia K, Shi P, Han X, Chen T, Tang H and Wang J 2016 Diagnostic value of miR-30d-5p and miR-125b-5p in acute myocardial infarction. Mol. Med. Rep. 14 184–194

    CAS  Article  Google Scholar 

  • Ko JH, Sethi G, Um JY, Shanmugam MK, Arfuso F, Kumar AP, Bishayee A and Ahn KS 2017 The Role of Resveratrol in Cancer Therapy. Int. J. Mol. Sci. 18 2589

    Article  Google Scholar 

  • Li X, Du N, Zhang Q, Li J, Chen X, Liu X, Hu Y, Qin W et al. 2014 MicroRNA-30d regulates cardiomyocyte pyroptosis by directly targeting foxo3a in diabetic cardiomyopathy. Cell Death Dis. 5 e1479

    CAS  Article  Google Scholar 

  • Li YG, Zhu W, Tao JP, Xin P, Liu MY, Li JB and Wei M 2013 Resveratrol protects cardiomyocytes from oxidative stress through SIRT1 and mitochondrial biogenesis signaling pathways. Biochem. Biophys. Res. Commun. 438 270–276

    CAS  Article  Google Scholar 

  • Ma L and Li Y 2015 SIRT1: role in cardiovascular biology. Clin Chim Acta 440 8–15

    CAS  Article  Google Scholar 

  • Meng X, Tan J, Li M, Song S, Miao Y and Zhang Q 2017 Sirt1: Role Under the Condition of Ischemia/Hypoxia. Cell. Mol. Neurobiol. 37 17–28

    CAS  Article  Google Scholar 

  • Michaille JJ, Piurowski V, Rigot B, Kelani H, Fortman EC and Tili E 2018 MiR-663, a MicroRNA linked with inflammation and cancer that is under the influence of resveratrol. Medicines 5 74

    CAS  Article  Google Scholar 

  • Moe GW and Marin-Garcia J 2016 Role of cell death in the progression of heart failure. Heart Fail. Rev. 21 157–167

    Article  Google Scholar 

  • QiNan W, XiaGuang G, XiaoTian L, WuQuan D, Ling Z and Bing C 2016 Par-4/NF-kappaB mediates the apoptosis of islet beta cells induced by glucolipotoxicity. J. Diabetes Res. 2016 4692478

    Article  Google Scholar 

  • Qiu G, Li X, Che X, Wei C, He S, Lu J, Jia Z, Pang K et al. 2015 SIRT1 is a regulator of autophagy: Implications in gastric cancer progression and treatment. FEBS Lett. 589 2034–2042

    CAS  Article  Google Scholar 

  • Teringova E and Tousek P 2017 Apoptosis in ischemic heart disease. J. Transl. Med. 15 87

    Article  Google Scholar 

  • Tili E and Michaille JJ 2016 Promiscuous effects of some phenolic natural products on inflammation at least in part arise from their ability to modulate the expression of global regulators, namely microRNAs. Molecules 21 1263

    Article  Google Scholar 

  • Wang J, Liew OW, Richards AM and Chen YT 2016 Overview of MicroRNAs in cardiac hypertrophy, fibrosis, and apoptosis. Int. J. Mol. Sci. 17 749

    Article  Google Scholar 

  • Wei C, Kim IK, Kumar S, Jayasinghe S, Hong N, Castoldi G, Catalucci D, Jones WK et al. 2013 NF-kappaB mediated miR-26a regulation in cardiac fibrosis. J. Cell. Physiol. 228 1433–1442

    CAS  Article  Google Scholar 

  • Wei C, Li L and Gupta S 2014 NF-kappaB-mediated miR-30b regulation in cardiomyocytes cell death by targeting Bcl-2. Mol. Cell. Biochem. 387 135–141

    CAS  Article  Google Scholar 

  • Wu Y, Meng X, Huang C and Li J 2015 Emerging role of silent information regulator 1 (SIRT1) in hepatocellular carcinoma: a potential therapeutic target. Tumour Biol. 36 4063–4074

    CAS  Article  Google Scholar 

  • Xiao J, Gao R, Bei Y, Zhou Q, Zhou Y, Zhang H, Jin M, Wei S et al. 2017 Circulating miR-30d predicts survival in patients with acute heart failure. Cell Physiol. Biochem. 41 865–874

    CAS  Article  Google Scholar 

  • Yang B, Ma S, Wang YB, Xu B, Zhao H, He YY, Li CW, Zhang J et al. 2016 Resveratrol exerts protective effects on anoxia/reoxygenation injury in cardiomyocytes via miR-34a/Sirt1 signaling pathway. Eur. Rev. Med. Pharmacol. Sci. 20 2734–2741

    CAS  PubMed  Google Scholar 

  • Yang Y, Duan W, Li Y, Jin Z, Yan J, Yu S and Yi D 2013 Novel role of silent information regulator 1 in myocardial ischemia. Circulation 128 2232–2240

    Article  Google Scholar 

  • Yu L, Liang H, Dong X, Zhao G, Jin Z, Zhai M, Yang Y, Chen W et al. 2015 Reduced silent information regulator 1 signaling exacerbates myocardial ischemia-reperfusion injury in type 2 diabetic rats and the protective effect of melatonin. J. Pineal. Res. 59 376–390

    CAS  Article  Google Scholar 

  • Zhang C, Lin G, Wan W, Li X, Zeng B, Yang B and Huang C 2012 Resveratrol, a polyphenol phytoalexin, protects cardiomyocytes against anoxia/reoxygenation injury via the TLR4/NF-kappaB signaling pathway. Int. J. Mol. Med. 29 557–563

    CAS  Article  Google Scholar 

  • Zhao F, Qu Y, Wang H, Huang L, Zhu J, Li S, Tong Y, Zhang L et al. 2017 The effect of miR-30d on apoptosis and autophagy in cultured astrocytes under oxygen-glucose deprivation. Brain Res. 1671 67–76

    CAS  Article  Google Scholar 

Download references

Author information

Author notes

  1. These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Cardiology, Jinan City People’s Hospital, Jinan, China

    Xia Han & Zeng Tang Zhang

  2. Department of Medicine, Laiwu People’s Hospital of Jinan, Laiwu, China

    Lu Zhang

  3. Department of Acupuncture, Jinan City People’s Hospital, Jinan, China

    Yingcai Liu

  4. Department of Neurology, Jinan City People’s Hospital, Jinan, China

    Menghai Wu

  5. Department of Pediatrics, Taihe Hospital, Hubei University of Medicine, Hubei, China

    Xingchao Li & Tao Li

Authors
  1. Xia Han
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Lu Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Yingcai Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Menghai Wu
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Xingchao Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Zeng Tang Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Tao Li
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Zeng Tang Zhang or Tao Li.

Additional information

Communicated by BJ RAO.

Corresponding editor: BJ Rao

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

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

Download citation

  • Received:

  • Accepted:

  • Published:

  • DOI: https://doi.org/10.1007/s12038-020-9997-9

Keywords

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