Long non-coding RNA Mirt2 relieves lipopolysaccharide-induced injury in PC12 cells by suppressing miR-429

Abstract

Long non-coding RNAs (lncRNAs) and microRNAs (miRNAs) play important roles in the pathogenesis of spinal cord injury (SCI). This study investigated the effects of lncRNA Mirt2 and miR-429 on lipopolysaccharide (LPS)-induced injuries in PC12 cells. Serum samples were collected from 36 patients with SCI and the healthy controls. The expression of lncRNA Mirt2 in serum samples was measured by qRT-PCR. The in vitro model of SCI was established by treating PC12 cells with LPS. The effects of lncRNA Mirt2 and miR-429 on the cell model were evaluated by CCK-8 assay, flow cytometry, western blot, qRT-PCR, and ELISA. Further, the activation of NF-κB and p38MAPK pathways was tested by western blot. LPS induced obvious cell injuries in PC12 cells, as cell viability was reduced, apoptosis rate was increased, caspase-3 and -9 were cleaved, and the release of TNF-α and IL-6 was induced. lncRNA Mirt2 was up-regulated in LPS-stimulated PC12 cells and serum samples derived from SCI patients. Overexpression of lncRNA Mirt2 protected PC12 cells against LPS-induced injuries. Further studies found that lncRNA Mirt2 acted as the molecular sponge of miR-429 and miR-34a-5p. lncRNA Mirt2 did not protect PC12 cells when miR-429 was overexpressed. Moreover, the inhibitory effects of lncRNA Mirt2 on NF-κB and p38MAPK pathways were abolished when miR-429 was overexpressed. lncRNA Mirt2 exerts protective effects in an in vitro model of SCI by down-regulating miR-429. This study shed light on the treatment of SCI by using the lncRNA-miRNA regulation network.

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

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Change history

  • 30 September 2020

    Editor's Note: Concerns have been raised about the integrity of the data reported in this article. This is currently being investigated. Further editorial action may be taken as appropriate once the investigation into the concerns is complete and all parties have been given an opportunity to respond in full.

References

  1. 1.

    Alizadeh A, Dyck SM, Karimi-Abdolrezaee S (2019) Traumatic spinal cord injury: an overview of pathophysiology, models and acute injury mechanisms. Front Neurol 10:282. https://doi.org/10.3389/fneur.2019.00282

    Article  PubMed  PubMed Central  Google Scholar 

  2. 2.

    Bartel DP (2004) MicroRNAs: genomics, biogenesis, mechanism, and function. Cell 116:281–297

    CAS  Article  Google Scholar 

  3. 3.

    Bernhard M, Gries A, Kremer P, Bottiger BW (2005) Spinal cord injury (SCI)--prehospital management. Resuscitation 66:127–139. https://doi.org/10.1016/j.resuscitation.2005.03.005

    Article  PubMed  Google Scholar 

  4. 4.

    Bethea JR, Castro M, Keane RW, Lee TT, Dietrich WD, Yezierski RP (1998) Traumatic spinal cord injury induces nuclear factor-kappaB activation. J Neurosci 18:3251–3260

    CAS  Article  Google Scholar 

  5. 5.

    Burke DA, Linden RD, Zhang YP, Maiste AC, Shields CB (2001) Incidence rates and populations at risk for spinal cord injury: a regional study. Spinal Cord 39:274–278. https://doi.org/10.1038/sj.sc.3101158

    CAS  Article  PubMed  Google Scholar 

  6. 6.

    Cao MX, Jiang YP, Tang YL, Liang XH (2017) The crosstalk between lncRNA and microRNA in cancer metastasis: orchestrating the epithelial-mesenchymal plasticity. Oncotarget 8:12472–12483. https://doi.org/10.18632/oncotarget.13957

    Article  PubMed  Google Scholar 

  7. 7.

    David S, Kroner A (2011) Repertoire of microglial and macrophage responses after spinal cord injury. Nat Rev Neurosci 12:388–399. https://doi.org/10.1038/nrn3053

    CAS  Article  PubMed  Google Scholar 

  8. 8.

    Deng G, Gao Y, Cen Z, He J, Cao B, Zeng G, Zong S (2018) miR-136-5p regulates the inflammatory response by targeting the IKKbeta/NF-kappaB/A20 pathway after spinal cord injury. Cell Physiol Biochem 50:512–524. https://doi.org/10.1159/000494165

    CAS  Article  PubMed  Google Scholar 

  9. 9.

    Di Gesualdo F, Capaccioli S, Lulli M (2014) A pathophysiological view of the long non-coding RNA world. Oncotarget 5:10976–10996. https://doi.org/10.18632/oncotarget.2770

    Article  PubMed  PubMed Central  Google Scholar 

  10. 10.

    Donnelly DJ, Popovich PG (2008) Inflammation and its role in neuroprotection, axonal regeneration and functional recovery after spinal cord injury. Exp Neurol 209:378–388. https://doi.org/10.1016/j.expneurol.2007.06.009

    CAS  Article  PubMed  Google Scholar 

  11. 11.

    Du M, Yuan L, Tan X, Huang D, Wang X, Zheng Z, Mao X, Li X, Yang L, Huang K, Zhang F, Wang Y, Luo X, Huang D, Huang K (2017) The LPS-inducible lncRNA Mirt2 is a negative regulator of inflammation. Nat Commun 8:2049. https://doi.org/10.1038/s41467-017-02229-1

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  12. 12.

    Fu S, Zhang J (2018) Knockdown of miR-429 attenuates abeta-induced neuronal damage by targeting SOX2 and BCL2 in mouse cortical neurons. 43:2240–2251. https://doi.org/10.1007/s11064-018-2643-3

  13. 13.

    Genovese T, Mazzon E, Crisafulli C, Di Paola R, Muia C, Esposito E, Bramanti P, Cuzzocrea S (2008) TNF-alpha blockage in a mouse model of SCI: evidence for improved outcome. Shock (Augusta, Ga) 29:32–41. https://doi.org/10.1097/shk.0b013e318059053a

    CAS  Article  Google Scholar 

  14. 14.

    Kaur H, Sarmah D, Saraf J, Vats K, Kalia K, Borah A, Yavagal DR, Dave KR, Ghosh Z, Bhattacharya P (2018) Noncoding RNAs in ischemic stroke: time to translate. Ann N Y Acad Sci 1421:19–36. https://doi.org/10.1111/nyas.13612

    CAS  Article  PubMed  Google Scholar 

  15. 15.

    Li F, Zhou MW (2019) MicroRNAs in contusion spinal cord injury: pathophysiology and clinical utility. https://doi.org/10.1007/s13760-019-01076-9

  16. 16.

    Li G, Chen T, Zhu Y, Xiao X, Bu J, Huang Z (2018) MiR-103 alleviates autophagy and apoptosis by regulating SOX2 in LPS-injured PC12 cells and SCI rats. Iran J Basic Med Sci 21:292–300. https://doi.org/10.22038/ijbms.2018.25980.6392

    Article  PubMed  PubMed Central  Google Scholar 

  17. 17.

    Li JW, Kuang Y, Chen L, Wang JF (2018) LncRNA ZNF667-AS1 inhibits inflammatory response and promotes recovery of spinal cord injury via suppressing JAK-STAT pathway. Eur Rev Med Pharmacol Sci 22:7614–7620. https://doi.org/10.26355/eurrev_201811_16375

    Article  PubMed  Google Scholar 

  18. 18.

    Li R, Yin F, Guo Y, Ruan Q, Zhu Q (2018) Angelica polysaccharide protects PC-12 cells from lipopolysaccharide-induced injury via down-regulating microRNA-223. Biomed Pharmacother 108:1320–1327. https://doi.org/10.1016/j.biopha.2018.09.147

    CAS  Article  PubMed  Google Scholar 

  19. 19.

    Lv HR (2017) lncRNA-Map2k4 sequesters miR-199a to promote FGF1 expression and spinal cord neuron growth. Biochem Biophys Res Commun 490:948–954. https://doi.org/10.1016/j.bbrc.2017.06.145

    CAS  Article  PubMed  Google Scholar 

  20. 20.

    Ni H, Jin W, Zhu T, Wang J, Yuan B, Jiang J, Liang W, Ma Z (2015) Curcumin modulates TLR4/NF-kappaB inflammatory signaling pathway following traumatic spinal cord injury in rats. J Spinal Cord Med 38:199–206. https://doi.org/10.1179/2045772313y.0000000179

    Article  PubMed  PubMed Central  Google Scholar 

  21. 21.

    Saklatvala J (2004) The p38 MAP kinase pathway as a therapeutic target in inflammatory disease. Curr Opin Pharmacol 4:372–377. https://doi.org/10.1016/j.coph.2004.03.009

    CAS  Article  PubMed  Google Scholar 

  22. 22.

    Wang J, Su B, Zhu H, Chen C, Zhao G (2016) Protective effect of geraniol inhibits inflammatory response, oxidative stress and apoptosis in traumatic injury of the spinal cord through modulation of NF-kappaB and p38 MAPK. Exp Ther Med 12:3607–3613. https://doi.org/10.3892/etm.2016.3850

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  23. 23.

    Wang Y, Zhao X, Xie H (2019) Quality of life and its predictors in people with traumatic spinal cord injury in mainland China. Spinal Cord. https://doi.org/10.1038/s41393-019-0279-z

  24. 24.

    Xiao J, Tang J, Chen Q, Tang D, Liu M, Luo M, Wang Y, Wang J, Zhao Z, Tang C, Wang D, Mo Z (2015) miR-429 regulates alveolar macrophage inflammatory cytokine production and is involved in LPS-induced acute lung injury. Biochem J 471:281–291. https://doi.org/10.1042/bj20131510

    CAS  Article  PubMed  Google Scholar 

  25. 25.

    Yan XT, Zhao Y, Cheng XL, He XH, Wang Y, Zheng WZ, Chen H, Wang YL (2018) Inhibition of miR-200b/miR-429 contributes to neuropathic pain development through targeting zinc finger E box binding protein-1. J Cell Physiol 233:4815–4824. https://doi.org/10.1002/jcp.26284

    CAS  Article  PubMed  Google Scholar 

  26. 26.

    Yang G, Tang WY (2017) Resistance of interleukin-6 to the extracellular inhibitory environment promotes axonal regeneration and functional recovery following spinal cord injury. Int J Mol Med 39:437–445. https://doi.org/10.3892/ijmm.2017.2848

    CAS  Article  PubMed  Google Scholar 

  27. 27.

    Zhang B, Li H, Li D, Sun H, Li M, Hu H (2019) Long noncoding RNA Mirt2 upregulates USP10 expression to suppress hepatic steatosis by sponging miR-34a-5p. Gene 700:139–148. https://doi.org/10.1016/j.gene.2019.02.096

    CAS  Article  PubMed  Google Scholar 

  28. 28.

    Zhang H, Wang Y (2016) Identification of molecular pathway changes after spinal cord injury by microarray analysis. J Orthop Surg Res 11:101. https://doi.org/10.1186/s13018-016-0437-3

    Article  PubMed  PubMed Central  Google Scholar 

  29. 29.

    Zhang Y, Cruickshanks N, Pahuski M, Yuan F, Dutta A, Schiff D, Purow B, Abounader R (2017) Noncoding RNAs in glioblastoma. In: De Vleeschouwer S (ed) Glioblastoma. Codon Publications Copyright. The Authors, Brisbane (AU). https://doi.org/10.15586/codon.glioblastoma.2017.ch6

    Google Scholar 

  30. 30.

    Zhao J, Wang L, Li Y (2017) Electroacupuncture alleviates the inflammatory response via effects on M1 and M2 macrophages after spinal cord injury. Acupunct Med 35:224–230. https://doi.org/10.1136/acupmed-2016-011107

    Article  PubMed  Google Scholar 

  31. 31.

    Zhou HJ, Wang LQ, Wang DB, Yu JB, Zhu Y, Xu QS, Zheng XJ, Zhan RY (2018) Long noncoding RNA MALAT1 contributes to inflammatory response of microglia following spinal cord injury via the modulation of a miR-199b/IKKbeta/NF-kappaB signaling pathway. Am J Physiol Cell Physiol 315:C52–c61. https://doi.org/10.1152/ajpcell.00278.2017

    CAS  Article  PubMed  Google Scholar 

Download references

Author information

Affiliations

Authors

Contributions

Haibo Li and Yu Xu are co-first authors.

Corresponding author

Correspondence to Huawei Ni.

Ethics declarations

Written consent was obtained from each adult volunteer before serum collection. The Ethics Committee of Affiliated Hospital of Hangzhou Normal University granted approval of this study.

Conflict of interest

The authors declare that they have no conflict of interest.

Additional information

Publisher’s note

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

Haibo Li and Yu Xu co-first authors

Statement

Shaoxing People’s Hospital (Shaoxing Hospital of Zhejiang University), Shenzhen University General Hospital, and The Affiliated Hospital of Hangzhou Normal University did not provide our authors the institutional email addresses. So we can only use our personal email address to submit the paper. We confirm that they are not fake email addresses. The authors can be reached at their email addresses.

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Li, H., Xu, Y., Wang, G. et al. Long non-coding RNA Mirt2 relieves lipopolysaccharide-induced injury in PC12 cells by suppressing miR-429. J Physiol Biochem 75, 403–413 (2019). https://doi.org/10.1007/s13105-019-00691-7

Download citation

Keywords

  • Mirt2
  • Spinal cord injury
  • miR-429
  • Inflammation
  • Apoptosis