Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

lncRNA NEAT1 aggravates sepsis-induced lung injury by regulating the miR-27a/PTEN axis

Abstract

Sepsis is an acute inflammatory reaction and a cause of acute respiratory distress syndrome (ARDS). In the present study, we explored the roles and underlying mechanism of the lncRNA Nuclear enriched abundant transcript 1 (NEAT1) in ARDS. The expression levels of genes, proteins and pro-inflammatory cytokines in patients with ARDS, LPS-stimulated cells and septic mouse models were quantified using qPCR, western blotting and ELISA assays, respectively. The molecular targeting relationship was validated by conducting a dual-luciferase reporter assay. Cell proliferation was assessed using the Cell Counting Kit-8 (CCK-8) assay. The cell cycle phase was determined by flow cytometry assay. The expression levels of NEAT1 and pro-inflammatory cytokines were higher in patients with ARDS and septic models than in controls. Knockdown of NEAT1 significantly increased cell proliferation and cycle progression and prolonged mouse survival in vitro and in vivo. Mechanistically, miR-27a was identified as a downstream target of NEAT1 and directly inhibited PTEN expression. Further rescue experiments revealed that inhibition of miR-27a impeded the promoting effects of NEAT1 silence on cell proliferation and cycle progression, whereas inhibition of PTEN markedly weakened the inhibitory effects of NEAT1 overexpression on cell proliferation and cycle progression. Altogether, our study revealed that NEAT1 plays a promoting role in the progression of ARDS via the NEAT1/miR-27a/PTEN regulatory network, providing new insight into the pathologic mechanism behind ARDS.

Access options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

Fig. 1: The expression levels of NEAT1 and inflammatory cytokines in ARDS patients and LPS-induced cells.
Fig. 2: Knockdown of NEAT1 attenuated LPS-induced lung cell injury.
Fig. 3: miR-27a was a downstream target of NEAT1.
Fig. 4: miR-27a reversed the effects of NEAT1 on the cell cycle and inflammatory response.
Fig. 5: PTEN functioned as an effector of miR-27a.
Fig. 6: PTEN participated in the regulatory effect of NEAT1 on LPS-induced cell injury.
Fig. 7: The role of NEAT1 knockdown in CLP-induced lung injury in mice.
Fig. 8

Data availability

The datasets used or analysed during the current study are available from the corresponding author on reasonable request.

References

  1. 1.

    Candel FJ, Borges Sa,M, Belda S, Bou G, Del Pozo JL, Estrada O, et al. Current aspects in sepsis approach. Turning things around. Rev Esp Quimioter. 2018;31:298–315.

    CAS  PubMed  PubMed Central  Google Scholar 

  2. 2.

    Schlichting D, McCollam JS. Recognizing and managing severe sepsis: a common and deadly threat. South Med J. 2007;100:594–600.

    Article  Google Scholar 

  3. 3.

    Yealy DM, Huang DT, Delaney A, Knight M, Randolph AG, Daniels R, et al. Recognizing and managing sepsis: what needs to be done? BMC Med. 2015;13:98.

    Article  Google Scholar 

  4. 4.

    Zhao H, Zhao M, Wang Y, Li F, Zhang Z. Glycyrrhizic acid prevents sepsis-induced acute lung injury and mortality in rats. J Histochem Cytochem. 2016;64:125–37.

    CAS  Article  Google Scholar 

  5. 5.

    Blumhagen RZ, Hedin BR, Malcolm KC, Burnham EL, Moss M, Abraham E, et al. Alternative pre-mRNA splicing of Toll-like receptor signaling components in peripheral blood mononuclear cells from patients with ARDS. Am J Physiol Lung Cell Mol Physiol. 2017;313:L930–l939.

    Article  Google Scholar 

  6. 6.

    Monaghan SF, Chung CS, Chen Y, Lomas-Neira J, Fairbrother WG, Heffernan DS, et al. Soluble programmed cell death receptor-1 (sPD-1): a potential biomarker with anti-inflammatory properties in human and experimental acute respiratory distress syndrome (ARDS). J Transl Med. 2016;14:312.

    Article  Google Scholar 

  7. 7.

    Liu X, Xiao ZD, Han L, Zhang J, Lee SW, Wang W, et al. LncRNA NBR2 engages a metabolic checkpoint by regulating AMPK under energy stress. Nat Cell Biol. 2016;18:431–42.

    CAS  Article  Google Scholar 

  8. 8.

    Chen Y, Lin Y, Bai Y, Cheng D, Bi Z. A long noncoding RNA (lncRNA)-associated competing endogenous RNA (ceRNA) network identifies eight lncRNA biomarkers in patients with osteoarthritis of the knee. Med Sci Monit. 2019;25:2058–65.

    CAS  Article  Google Scholar 

  9. 9.

    Wang P, Liu G, Xu W, Liu H, Bu Q, Sun D. Long noncoding RNA H19 inhibits cell viability, migration, and invasion via downregulation of IRS-1 in thyroid cancer cells. Technol Cancer Res Treat. 2017;16:1102–12.

    CAS  Article  Google Scholar 

  10. 10.

    Aune TM, Spurlock CF 3rd. Long non-coding RNAs in innate and adaptive immunity. Virus Res. 2016;212:146–60.

    CAS  Article  Google Scholar 

  11. 11.

    He F, Zhang C, Huang Q. Long noncoding RNA nuclear enriched abundant transcript 1/miRNA-124 axis correlates with increased disease risk, elevated inflammation, deteriorative disease condition, and predicts decreased survival of sepsis. Medicine. 2019;98:e16470.

    CAS  Article  Google Scholar 

  12. 12.

    Yong H, Wu G, Chen J, Liu X, Bai Y, Tang N, et al. lncRNA MALAT1 accelerates skeletal muscle cell apoptosis and inflammatory response in sepsis by decreasing BRCA1 expression by recruiting EZH2. Mol Ther Nucleic Acids. 2020;19:97–108.

    CAS  Article  Google Scholar 

  13. 13.

    Klec C, Prinz F, Pichler M. Involvement of the long noncoding RNA NEAT1 in carcinogenesis. Mol Oncol. 2019;13:46–60.

    CAS  Article  Google Scholar 

  14. 14.

    Zhang CC, Niu F. LncRNA NEAT1 promotes inflammatory response in sepsis-induced liver injury via the Let-7a/TLR4 axis. Int Immunopharmacol. 2019;75:105731.

    CAS  Article  Google Scholar 

  15. 15.

    Wang SM, Liu GQ, Xian HB, Si JL, Qi SX, Yu YP. LncRNA NEAT1 alleviates sepsis-induced myocardial injury by regulating the TLR2/NF-kappaB signaling pathway. Eur Rev Med Pharmacol Sci. 2019;23:4898–907.

    PubMed  Google Scholar 

  16. 16.

    Huang S, Huang Z, Luo Q, Qing C. The expression of lncRNA NEAT1 in human tuberculosis and its antituberculosis effect. Biomed Res Int. 2018;2018:9529072.

    PubMed  PubMed Central  Google Scholar 

  17. 17.

    Shen Y, Yu J, Jing Y, Zhang J. MiR-106a aggravates sepsis-induced acute kidney injury by targeting THBS2 in mice model. Acta Cir Bras. 2019;34:e201900602.

    Article  Google Scholar 

  18. 18.

    Ling L, Lu HT, Wang HF, Shen MJ, Zhang HB. MicroRNA-203 acts as a potent suppressor in septic shock by alleviating lung injury via inhibition of VNN1. Kidney Blood Press Res. 2019;44:565–82.

    CAS  Article  Google Scholar 

  19. 19.

    Lin Q, Gao Z, Alarcon RM, Ye J, Yun Z. A role of miR-27 in the regulation of adipogenesis. FEBS J. 2009;276:2348–58.

    CAS  Article  Google Scholar 

  20. 20.

    Xie W, Li L, Zhang M, Cheng HP, Gong D, Lv YC, et al. MicroRNA-27 prevents atherosclerosis by suppressing lipoprotein lipase-induced lipid accumulation and inflammatory response in apolipoprotein E knockout mice. PLoS ONE. 2016;11:e0157085.

    Article  Google Scholar 

  21. 21.

    Zhang M, Wu JF, Chen WJ, Tang SL, Mo ZC, Tang YY, et al. MicroRNA-27a/b regulates cellular cholesterol efflux, influx and esterification/hydrolysis in THP-1 macrophages. Atherosclerosis. 2014;234:54–64.

    CAS  Article  Google Scholar 

  22. 22.

    Cheng Y, Kuang W, Hao Y, Zhang D, Lei M, Du L, et al. Downregulation of miR-27a* and miR-532-5p and upregulation of miR-146a and miR-155 in LPS-induced RAW264.7 macrophage cells. Inflammation. 2012;35:1308–13.

    CAS  Article  Google Scholar 

  23. 23.

    Yang Q, Zhang D, Li Y, Li Y, Li Y. Paclitaxel alleviated liver injury of septic mice by alleviating inflammatory response via microRNA-27a/TAB3/NF-kappaB signaling pathway. Biomed Pharmacother. 2018;97:1424–33.

    CAS  Article  Google Scholar 

  24. 24.

    Vasilescu C, Dragomir M, Tanase M, Giza D, Purnichescu-Purtan R, Chen M, et al. Circulating miRNAs in sepsis-A network under attack: an in-silico prediction of the potential existence of miRNA sponges in sepsis. PLoS ONE. 2017;12:e0183334.

    Article  Google Scholar 

  25. 25.

    Lee S, Nakahira K, Dalli J, Siempos II, Norris PC, Colas RA, et al. NLRP3 inflammasome deficiency protects against microbial sepsis via increased lipoxin B4 synthesis. Am J Respir Crit Care Med. 2017;196:713–26.

    CAS  Article  Google Scholar 

  26. 26.

    Mohnle P, Hirschberger S, Hinske LC, Briegel J, Hubner M, Weis S, et al. MicroRNAs 143 and 150 in whole blood enable detection of T-cell immunoparalysis in sepsis. Mol Med. 2018;24:54.

    CAS  Article  Google Scholar 

  27. 27.

    Neudecker, V, Brodsky, KS, Clambey, ET, Schmidt, EP, Packard, TA, Davenport, B et al. Neutrophil transfer of miR-223 to lung epithelial cells dampens acute lung injury in mice. Sci Transl Med. 9, eaah5360 (2017).

  28. 28.

    Bao, X, Zhang, Q, Liu, N, Zhuang, S, Li, Z, Meng, Q et al. Characteristics of circular RNA expression of pulmonary macrophages in mice with sepsis-induced acute lung injury. J Cell Mol Med https://doi.org/10.1111/jcmm.14577 (2019).

  29. 29.

    Liu W, Liu K, Zhang S, Shan L, Tang J. Tetramethylpyrazine showed therapeutic effects on sepsis-induced acute lung injury in rats by inhibiting endoplasmic reticulum stress protein kinase RNA-like endoplasmic reticulum kinase (PERK) signaling-induced apoptosis of pulmonary microvascular endothelial cells. Med Sci Monit. 2018;24:1225–31.

    CAS  Article  Google Scholar 

  30. 30.

    Wang L, Liu J, Xie W, Li G, Yao L, Zhang R, et al. Overexpression of MALAT1 relates to lung injury through sponging miR-425 and promoting cell apoptosis during ARDS. Can Respir J. 2019;2019:1871394.

    PubMed  PubMed Central  Google Scholar 

  31. 31.

    Wang Y, Fu X, Yu B, Ai F. Long non-coding RNA THRIL predicts increased acute respiratory distress syndrome risk and positively correlates with disease severity, inflammation, and mortality in sepsis patients. J Clin Lab Anal. 2019;33:e22882.

    PubMed  PubMed Central  Google Scholar 

  32. 32.

    Chen Y, Qiu J, Chen B, Lin Y, Chen Y, Xie G, et al. Long non-coding RNA NEAT1 plays an important role in sepsis-induced acute kidney injury by targeting miR-204 and modulating the NF-kappaB pathway. Int Immunopharmacol. 2018;59:252–60.

    CAS  Article  Google Scholar 

  33. 33.

    Huang S, Qian K, Zhu Y, Huang Z, Luo Q, Qing C. Diagnostic value of the lncRNA NEAT1 in peripheral blood mononuclear cells of patients with sepsis. Dis Markers. 2017;2017:7962836.

    PubMed  PubMed Central  Google Scholar 

  34. 34.

    Prinz, F, Kapeller, A, Pichler, M, & Klec, C. The implications of the long non-coding RNA NEAT1 in non-cancerous diseases. Int J Mol Sci. 20, 627 (2019).

  35. 35.

    Li W, Qiu X, Liu J, Han Y, Wei D, Ji G, et al. miR-27a protects against acute lung injury in LPS-treated mice by inhibiting NF-kappaB-mediated inflammatory response. Int J Clin Exp Pathol. 2018;11:2980–9.

    PubMed  PubMed Central  Google Scholar 

  36. 36.

    Xie N, Cui H, Banerjee S, Tan Z, Salomao R, Fu M, et al. miR-27a regulates inflammatory response of macrophages by targeting IL-10. J Immunol. 2014;193:327–34.

    CAS  Article  Google Scholar 

  37. 37.

    Ju M, Liu B, He H, Gu Z, Liu Y, Su Y, et al. MicroRNA-27a alleviates LPS-induced acute lung injury in mice via inhibiting in fl ammation and apoptosis through modulating TLR4/MyD88/NF-kappaB pathway. Cell Cycle. 2018;17:2001–18.

    CAS  Article  Google Scholar 

  38. 38.

    Wang Z, Ruan Z, Mao Y, Dong W, Zhang Y, Yin N, et al. miR-27a is up regulated and promotes inflammatory response in sepsis. Cell Immunol. 2014;290:190–5.

    CAS  Article  Google Scholar 

  39. 39.

    Wang Y, Zhang X, Tian J, Liu G, Li X, Shen D. Sevoflurane alleviates LPSinduced acute lung injury via the microRNA27a3p/TLR4/MyD88/NFkappaB signaling pathway. Int J Mol Med. 2019;44:479–90.

    CAS  PubMed  PubMed Central  Google Scholar 

  40. 40.

    Brandmaier A, Hou SQ, Shen WH. Cell cycle control by PTEN. J Mol Biol. 2017;429:2265–77.

    CAS  Article  Google Scholar 

  41. 41.

    Sachdeva K, Do DC, Zhang Y, Hu X, Chen J, Gao P. Environmental exposures and asthma development: autophagy, mitophagy, and cellular senescence. Front Immunol. 2019;10:2787.

    CAS  Article  Google Scholar 

  42. 42.

    Zhou M, Fang H, Du M, Li C, Tang R, Liu H, et al. The modulation of regulatory T cells via HMGB1/PTEN/β-catenin axis in LPS induced acute lung injury. Front Immunol. 2019;10:1612.

    CAS  Article  Google Scholar 

  43. 43.

    Wu J, Sun Z, Sun H, Li Y. MicroRNA27a promotes tumorigenesis via targeting AKT in triple negative breast cancer. Mol Med Rep. 2018;17:562–70.

    CAS  PubMed  Google Scholar 

  44. 44.

    Liu, L, Chen, J, Zhang, X, Cui, X, Qiao, N, Zhang, Y et al. The protective effect of PPARgamma in sepsis-induced acute lung injury via inhibiting PTEN/beta-catenin pathway. Biosci Rep. 40, BSR20192639 (2020).

Download references

Funding

This work is supported by special funds for basic scientific research business expenses of the central public welfare research institutes of the Chinese Academy of Medical Sciences (No. 2019PT32000), Doctor Foundation of Guizhou Provincial People’s Hospital (GZSYBS [2019]04).

Author information

Affiliations

Authors

Contributions

XL: study concepts, definition of intellectual content and statistical analysis; XYZ: study concepts, manuscript preparation and editing; YJN: data acquisition and definition of intellectual content; GHL and QZ: literature research and experimental studies; XF: data analysis and statistical analysis; SY: data analysis; QHZ: study design, manuscript preparation and editing and manuscript review; JQL: guarantor of integrity of the entire study, study concepts and design and manuscript review. All authors read and approved the final manuscript.

Corresponding authors

Correspondence to Qing-Hua Zhao or Jian-Quan Li.

Ethics declarations

Competing interests

The authors declare no competing interests.

Ethics approval

All protocol was approved by the Ethics Committee of Guizhou Provincial People’s Hospital and conducted according to the Declaration of Helsinki.

Informed consent

All patients were provided with informed consent.

Additional information

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

Supplementary information

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Lv, X., Zhang, XY., Zhang, Q. et al. lncRNA NEAT1 aggravates sepsis-induced lung injury by regulating the miR-27a/PTEN axis. Lab Invest 101, 1371–1381 (2021). https://doi.org/10.1038/s41374-021-00620-7

Download citation

Search

Quick links