Skip to main content
SpringerLink
Log in

The Mitochondrial-Derived Peptide MOTS-c Attenuates Oxidative Stress Injury and the Inflammatory Response of H9c2 Cells Through the Nrf2/ARE and NF-κB Pathways

  • Original Article
  • Published:
Cardiovascular Engineering and Technology Aims and scope Submit manuscript

Abstract

Aim

Oxidative stress and the inflammatory response contribute to the progression of cardiovascular disease. The present study aimed to investigate whether the mitochondrial-derived peptide MOTS-c could alleviate H2O2-induced oxidative stress and inflammatory status in H9c2 cells through activation of nuclear factor erythroid 2-related Factor 2 (Nrf2)/antioxidative response element (ARE) and inhibition of the NF-κB pathway.

Methods

Rat H9c2 cardiomyocytes were obtained, and 10, 20 or 50 μM MOTS-c was pretreated for 24 h before treatment with H2O2. Then, the cell was treated with 100 μM H2O2 for 1 h to induce oxidative stress. An inhibition model of sh-Nrf2 was constructed via a lentivirus expression system, and an activation model of NF-κB was achieved using phorbol 12-myristate-13-acetate (PMA). Cell viability was determined using a Cell Counting kit-8 assay. Relative measurement of relative protein and mRNA expression used western blotting and qRT-PCR, respectively. Intracellular reactive oxygen species (ROS) levels were detected using dichlorodihydrofluorescein diacetate, and malondialdehyde (MDA) and superoxide dismutase (SOD) levels were determined via commercial kits. The protein expression and distribution in the cells were visualized by immunofluorescence analysis. Enzyme-linked immunosorbent assay was used to detect the levels of inflammatory cytokines, including TNF-α, IL-6 and IL-1β.

Results

We found that H2O2 treatment significantly decreased cell viability and the level of SOD, increased the levels of ROS and MDA, and upregulated the expression of inflammatory cytokines, including TNF-α, IL-6 and IL-1β, in H9c2 cells. The expression levels of Nrf2, HO-1 and NQO-1 were significantly downregulated in the H2O2, while the phosphorylation of NF-κBp65 was promoted by H2O2. However, pretreatment with MOTS-c significantly reversed H2O2-induced damage in H9c2 cells. Moreover, both inhibition of the Nrf2/ARE pathway and activation of the NF-κB pathway significantly decreased the effects of MOTS-c, suggesting that MOTS-c might play a role in alleviating oxidative damage via the Nrf2/ARE and NF-κB pathways.

Conclusions

Our investigation indicated that MOTS-c could protect against H2O2-induced inflammation and oxidative stress in H9c2 cells by inhibiting NF-κB and activating the Nrf2/ARE pathways.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

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

Instant access to the full article PDF.

Institutional subscriptions

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5

Similar content being viewed by others

References

  1. Bundy, J. D., and J. He. Hypertension and related cardiovascular disease burden in China. Ann. Glob. Health. 82(2):227–233, 2016.

    Article  PubMed  Google Scholar 

  2. Chang, H., C. Li, K. Huo, Q. Wang, L. Lu, Q. Zhang, et al. Luteolin prevents H2O2-induced apoptosis in H9C2 cells through modulating Akt-P53/Mdm2 signaling pathway. Biomed. Res. Int. 2016:5125836, 2016.

    Article  PubMed  PubMed Central  Google Scholar 

  3. Chen, Q. M., and A. J. Maltagliati. Nrf2 at the heart of oxidative stress and cardiac protection. Physiol. Genomics. 50(2):77–97, 2018.

    Article  CAS  PubMed  Google Scholar 

  4. Del Olmo-Garcia, M. I., and J. F. Merino-Torres. GLP-1 receptor agonists and cardiovascular disease in patients with type 2 diabetes. J. Diabetes Res. 2018:4020492, 2018.

    PubMed  PubMed Central  Google Scholar 

  5. Dias, A. T., A. S. Cintra, J. C. Frossard, Z. Palomino, D. E. Casarini, I. B. Gomes, et al. Inhibition of phosphodiesterase 5 restores endothelial function in renovascular hypertension. J. Transl. Med. 12:250, 2014.

    Article  PubMed  PubMed Central  Google Scholar 

  6. Ebrahimi, B., J. A. Crane, B. E. Knudsen, S. I. Macura, J. P. Grande, and L. O. Lerman. Evolution of cardiac and renal impairment detected by high-field cardiovascular magnetic resonance in mice with renal artery stenosis. J. Cardiovasc. Magn. Reson. 15(1):98, 2013.

    Article  PubMed  PubMed Central  Google Scholar 

  7. Fang, M., W. H. Zhong, W. L. Song, Y. Y. Deng, D. M. Yang, B. Xiong, et al. Ulinastatin ameliorates pulmonary capillary endothelial permeability induced by sepsis through protection of tight junctions via inhibition of TNF-α and Related pathways. Front. Pharmacol. 9:823, 2018.

    Article  PubMed  PubMed Central  Google Scholar 

  8. Gaspari, T., H. Liu, I. Welungoda, Y. Hu, R. E. Widdop, L. B. Knudsen, et al. A GLP-1 receptor agonist liraglutide inhibits endothelial cell dysfunction and vascular adhesion molecule expression in an ApoE-/- mouse model. Diab. Vasc. Dis. Res. 8(2):117–124, 2011.

    Article  PubMed  Google Scholar 

  9. Ghorpade, D. S., L. Ozcan, Z. Zheng, S. M. Nicoloro, Y. Shen, E. Chen, et al. Hepatocyte-secreted DPP4 in obesity promotes adipose inflammation and insulin resistance. Nature. 555(7698):673–677, 2018.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Hu, B. T., and W. Z. Chen. MOTS-c improves osteoporosis by promoting osteogenic differentiation of bone marrow mesenchymal stem cells via TGF-β/Smad pathway. Eur. Rev. Med. Pharmacol. Sci. 22(21):7156–7163, 2018.

    PubMed  Google Scholar 

  11. Li, D., X. Wang, Q. Huang, S. Li, Y. Zhou, and Z. Li. Cardioprotection of CAPE-oNO(2) against myocardial ischemia/reperfusion induced ROS generation via regulating the SIRT1/eNOS/NF-κB pathway in vivo and in vitro. Redox Biol. 15:62–73, 2018.

    Article  CAS  PubMed  Google Scholar 

  12. Liu, C., E. K. Gidlund, A. Witasp, A. R. Qureshi, M. Söderberg, A. Thorell, et al. Reduced skeletal muscle expression of mitochondrial-derived peptides humanin and MOTS-C and Nrf2 in chronic kidney disease. Am. J. Physiol. Renal. Physiol. 317(5):F1122–F1131, 2019.

    Article  CAS  PubMed  Google Scholar 

  13. Muzumdar, R. H., D. M. Huffman, G. Atzmon, C. Buettner, L. J. Cobb, S. Fishman, et al. Humanin: a novel central regulator of peripheral insulin action. PLoS ONE. 4(7):e6334, 2009.

    Article  PubMed  PubMed Central  Google Scholar 

  14. North, B. J., and D. A. Sinclair. The intersection between aging and cardiovascular disease. Circ. Res. 110(8):1097–1108, 2012.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Priestley, J. R. C., K. E. Fink, J. M. McCord, and J. H. Lombard. NRF2 activation with Protandim attenuates salt-induced vascular dysfunction and microvascular rarefaction. Microcirculation. 26(7):e12575, 2019.

    Article  PubMed  PubMed Central  Google Scholar 

  16. Qin, Q., S. Delrio, J. Wan, R. Jay Widmer, P. Cohen, L. O. Lerman, et al. Downregulation of circulating MOTS-c levels in patients with coronary endothelial dysfunction. Int. J. Cardiol. 254:23–27, 2018.

    Article  PubMed  Google Scholar 

  17. Qiu, Z., Y. He, H. Ming, S. Lei, Y. Leng, and Z. Y. Xia. Lipopolysaccharide (LPS) aggravates high glucose- and hypoxia/reoxygenation-induced injury through activating ROS-dependent NLRP3 inflammasome-mediated pyroptosis in H9C2 cardiomyocytes. J. Diabetes Res. 2019:8151836, 2019.

    Article  PubMed  PubMed Central  Google Scholar 

  18. Ran, N., C. Lin, L. Leng, G. Han, M. Geng, Y. Wu, et al. MOTS-c promotes phosphorodiamidate morpholino oligomer uptake and efficacy in dystrophic mice. EMBO Mol. Med. 13(2):e12993, 2021.

    Article  CAS  PubMed  Google Scholar 

  19. Sajadimajd, S., and M. Khazaei. Oxidative stress and cancer: the role of Nrf2. Curr. Cancer Drug Targets. 18(6):538–557, 2018.

    Article  CAS  PubMed  Google Scholar 

  20. Simonson, B., and S. Das. MicroRNA therapeutics: the next magic bullet? Mini Rev. Med. Chem. 15(6):467–474, 2015.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Sternik, L., S. Samee, H. V. Schaff, K. J. Zehr, L. O. Lerman, D. R. Holmes, et al. C-reactive protein relaxes human vessels in vitro. Arterioscler. Thromb. Vasc. Biol. 22(11):1865–1868, 2002.

    Article  CAS  PubMed  Google Scholar 

  22. Steven, S., K. Frenis, M. Oelze, S. Kalinovic, M. Kuntic, M. T. Bayo Jimenez, et al. Vascular inflammation and oxidative stress: major triggers for cardiovascular disease. Oxid. Med. Cell Longev. 2019:7092151, 2019.

    Article  PubMed  PubMed Central  Google Scholar 

  23. Sun, H. J., Z. Y. Wu, X. W. Nie, and J. S. Bian. Role of endothelial dysfunction in cardiovascular diseases: the link between inflammation and hydrogen sulfide. Front. Pharmacol. 10:1568, 2019.

    Article  CAS  PubMed  Google Scholar 

  24. Tahara, A., E. Kurosaki, M. Yokono, D. Yamajuku, R. Kihara, Y. Hayashizaki, et al. Effects of SGLT2 selective inhibitor ipragliflozin on hyperglycemia, hyperlipidemia, hepatic steatosis, oxidative stress, inflammation, and obesity in type 2 diabetic mice. Eur. J. Pharmacol. 715(1–3):246–255, 2013.

    Article  CAS  PubMed  Google Scholar 

  25. Wei, M., L. Gan, Z. Liu, L. Liu, J. R. Chang, D. C. Yin, et al. Mitochondrial-derived peptide MOTS-c attenuates vascular calcification and secondary myocardial remodeling via adenosine monophosphate-activated protein kinase signaling pathway. Cardiorenal. Med. 10(1):42–50, 2020.

    Article  CAS  PubMed  Google Scholar 

  26. Wojciechowska, A., A. Braniewska, and K. Kozar-Kamińska. MicroRNA in cardiovascular biology and disease. Adv. Clin. Exp. Med. 26(5):865–874, 2017.

    Article  PubMed  Google Scholar 

  27. Xu, W., J. Chen, J. Lin, D. Liu, L. Mo, W. Pan, et al. Exogenous H2S protects H9c2 cardiac cells against high glucose-induced injury and inflammation by inhibiting the activation of the NF-κB and IL-1β pathways. Int. J. Mol. Med. 35(1):177–186, 2015.

    Article  CAS  PubMed  Google Scholar 

  28. Yan, Z., S. Zhu, H. Wang, L. Wang, T. Du, Z. Ye, et al. MOTS-c inhibits osteolysis in the Mouse Calvaria by affecting osteocyte-osteoclast crosstalk and inhibiting inflammation. Pharmacol. Res. 147:104381, 2019.

    Article  CAS  PubMed  Google Scholar 

  29. Yen, K., C. Lee, H. Mehta, and P. Cohen. The emerging role of the mitochondrial-derived peptide humanin in stress resistance. J. Mol. Endocrinol. 50(1):R11–R19, 2013.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. Zhang, G., X. Zou, Y. Huang, F. Wang, S. Miao, G. Liu, et al. Mesenchymal stromal cell-derived extracellular vesicles protect against acute kidney injury through anti-oxidation by enhancing Nrf2/ARE activation in rats. Kidney Blood Press. Res. 41(2):119–128, 2016.

    Article  CAS  PubMed  Google Scholar 

  31. Zhu, H., Y. Li, M. X. Wang, J. H. Wang, W. X. Du, and F. Zhou. Analysis of cardiovascular disease-related NF-κB-regulated genes and microRNAs in TNFα-treated primary mouse vascular endothelial cells. J. Zhejiang Univ. Sci. B. 20(10):803–815, 2019.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Funding

This work was supported by Funding of the Plan of Science and Technology on Medical and Health in Zhejiang Province (Nos. 2021KY1002, 2022KY312), Ningbo province public welfare project (No. 20211JCGY020364) and Natural Science Foundation of Ningbo (No. 2021J266).

Conflict of interest

The authors ensure that they have no conflict of interests.

Author information

Author notes

  1. Caijie Shen and Jian Wang are co-first authors.

Authors and Affiliations

  1. Department of Cardiovascular Medicine, Ningbo First Hospital, No. 59, Liuting Street, Haishu District, Ningbo, 315000, Zhejiang, People’s Republic of China

    Caijie Shen, Jian Wang, Mingjun Feng, Xiangfeng Du, Huimin Chu & Xiaomin Chen

  2. Department of Cardiovascular Medicine, The Second Affiliated Hospital of University of South China, Hengyang, 315000, Hunan, People’s Republic of China

    Jianye Peng

Authors
  1. Caijie Shen
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Jian Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Mingjun Feng
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Jianye Peng
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Xiangfeng Du
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Huimin Chu
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Xiaomin Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Huimin Chu or Xiaomin Chen.

Additional information

Associate Editor Igor Efimov oversaw the review of this article.

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Shen, C., Wang, J., Feng, M. et al. The Mitochondrial-Derived Peptide MOTS-c Attenuates Oxidative Stress Injury and the Inflammatory Response of H9c2 Cells Through the Nrf2/ARE and NF-κB Pathways. Cardiovasc Eng Tech 13, 651–661 (2022). https://doi.org/10.1007/s13239-021-00589-w

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s13239-021-00589-w

Keywords

Search

Navigation