open access

Vol 57, No 4 (2019)
Original paper
Submitted: 2019-09-27
Accepted: 2019-11-14
Published online: 2019-12-11
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MiR-539-5p alleviates sepsis-induced acute lung injury by targeting ROCK1

Li Meng1, Haohao Cao2, Chunhua Wan1, Lintao Jiang3
DOI: 10.5603/FHC.a2019.0019
·
Pubmed: 31825519
·
Folia Histochem Cytobiol 2019;57(4):168-178.
Affiliations
  1. Departement of Anesthesiology, The Affiliated Hospital of Hubei Provincial Government, Wuhan, China
  2. Departement of Critical Care Medicine, Wuhan Forth Hospital, Wuhan,Hubei Province, 430000, China
  3. Departement of Emergency, Central Hospital of Wuhan, Wuhan,Hubei Province,430000,China

open access

Vol 57, No 4 (2019)
ORIGINAL PAPERS
Submitted: 2019-09-27
Accepted: 2019-11-14
Published online: 2019-12-11

Abstract

Introduction. Sepsis-induced acute lung injury (ALI) is an inflammatory process involved with simultaneous production of inflammatory cytokines and chemokines. In this study, we investigated the regulatory role of miR-539-5p in sepsis-induced ALI using a mouse model of cecal ligation puncture (CLP) and an in vitro model of primary murine pulmonary microvascular endothelial cells (MPVECs).


Material and methods. Adult male C57BL/6 mice were intravenously injected with or without miR-539-5p agomir or scrambled control one week before CLP operation. MPVECs were transfected with miR-539-5p mimics or control mimics, followed by lipopolysaccharide (LPS) stimulation. ROCK1 was predicted and confirmed as a direct target of miR-539-5p using dual-luciferase reporter assay. In rescue experiment, MPVECs were co-transfected with lentiviral vector expressing ROCK1 (or empty vector) and miR-539-5p mimics 24 h before LPS treatment. The transcriptional activity of caspase-3, the apoptosis ratio, the levels of miR-539-5p, interleukin-1b (IL-1b), interleukin-6 (IL-6), and ROCK1 were assessed.


Results. Compared to sham group, mice following CLP showed pulmonary morphological abnormalities, elevated production of IL-1b and IL-6, and increased caspase-3 activity and apoptosis ratio in the lung. In MPVECs, LPS stimulation resulted in a significant induction of inflammatory cytokine levels and apoptosis compared to untreated cells. The overexpression of miR-539-5p in septic mice alleviated sepsis-induced pulmonary injury, apoptosis, and inflammation. MiR-539-5p also demonstrated anti-apoptotic and anti-inflammatory effect in LPS-treated MPVECs. The upregulation of ROCK1 in MPVECs recovered miR-539-5p-suppressed caspase-3 activity and proinflammatory cytokine production.


Conclusion. In conclusion, miR-539-5p alleviated sepsis-induced ALI via suppressing its downstream target ROCK1, suggesting a therapeutic potential of miR-539-5p for the management of sepsis-induced ALI.

Abstract

Introduction. Sepsis-induced acute lung injury (ALI) is an inflammatory process involved with simultaneous production of inflammatory cytokines and chemokines. In this study, we investigated the regulatory role of miR-539-5p in sepsis-induced ALI using a mouse model of cecal ligation puncture (CLP) and an in vitro model of primary murine pulmonary microvascular endothelial cells (MPVECs).


Material and methods. Adult male C57BL/6 mice were intravenously injected with or without miR-539-5p agomir or scrambled control one week before CLP operation. MPVECs were transfected with miR-539-5p mimics or control mimics, followed by lipopolysaccharide (LPS) stimulation. ROCK1 was predicted and confirmed as a direct target of miR-539-5p using dual-luciferase reporter assay. In rescue experiment, MPVECs were co-transfected with lentiviral vector expressing ROCK1 (or empty vector) and miR-539-5p mimics 24 h before LPS treatment. The transcriptional activity of caspase-3, the apoptosis ratio, the levels of miR-539-5p, interleukin-1b (IL-1b), interleukin-6 (IL-6), and ROCK1 were assessed.


Results. Compared to sham group, mice following CLP showed pulmonary morphological abnormalities, elevated production of IL-1b and IL-6, and increased caspase-3 activity and apoptosis ratio in the lung. In MPVECs, LPS stimulation resulted in a significant induction of inflammatory cytokine levels and apoptosis compared to untreated cells. The overexpression of miR-539-5p in septic mice alleviated sepsis-induced pulmonary injury, apoptosis, and inflammation. MiR-539-5p also demonstrated anti-apoptotic and anti-inflammatory effect in LPS-treated MPVECs. The upregulation of ROCK1 in MPVECs recovered miR-539-5p-suppressed caspase-3 activity and proinflammatory cytokine production.


Conclusion. In conclusion, miR-539-5p alleviated sepsis-induced ALI via suppressing its downstream target ROCK1, suggesting a therapeutic potential of miR-539-5p for the management of sepsis-induced ALI.

Get Citation

Keywords

mouse; acute lung injury; sepsis; inflammation; MPVEC cells; miRNA; ROCK1; caspase-3; cytokines

About this article
Title

MiR-539-5p alleviates sepsis-induced acute lung injury by targeting ROCK1

Journal

Folia Histochemica et Cytobiologica

Issue

Vol 57, No 4 (2019)

Article type

Original paper

Pages

168-178

Published online

2019-12-11

DOI

10.5603/FHC.a2019.0019

Pubmed

31825519

Bibliographic record

Folia Histochem Cytobiol 2019;57(4):168-178.

Keywords

mouse
acute lung injury
sepsis
inflammation
MPVEC cells
miRNA
ROCK1
caspase-3
cytokines

Authors

Li Meng
Haohao Cao
Chunhua Wan
Lintao Jiang

References (37)
  1. Sagy M, Al-Qaqaa Y, Kim P. Definitions and pathophysiology of sepsis. Curr Probl Pediatr Adolesc Health Care. 2013; 43(10): 260–263.
  2. Gando S. Microvascular thrombosis and multiple organ dysfunction syndrome. Crit Care Med. 2010; 38(2 Suppl): S35–S42.
  3. Levitt JE, Matthay MA. The utility of clinical predictors of acute lung injury: towards prevention and earlier recognition. Expert Rev Respir Med. 2010; 4(6): 785–797.
  4. Johnson ER, Matthay MA. Acute lung injury: epidemiology, pathogenesis, and treatment. J Aerosol Med Pulm Drug Deliv. 2010; 23(4): 243–252.
  5. Levitt JE, Matthay MA. Clinical review: Early treatment of acute lung injury--paradigm shift toward prevention and treatment prior to respiratory failure. Crit Care. 2012; 16(3): 223.
  6. Park WY, Goodman RB, Steinberg KP, et al. Cytokine balance in the lungs of patients with acute respiratory distress syndrome. Am J Respir Crit Care Med. 2001; 164(10 Pt 1): 1896–1903.
  7. Goodman RB, Strieter RM, Martin DP, et al. Inflammatory cytokines in patients with persistence of the acute respiratory distress syndrome. Am J Respir Crit Care Med. 1996; 154(3 Pt 1): 602–611.
  8. Martin TR, Nakamura M, Matute-Bello G. The role of apoptosis in acute lung injury. Crit Care Med. 2003; 31(4 Suppl): S184–S188.
  9. Perl M, Chung CS, Perl U, et al. Fas-induced pulmonary apoptosis and inflammation during indirect acute lung injury. Am J Respir Crit Care Med. 2007; 176(6): 591–601.
  10. Kitamura Y, Hashimoto S, Mizuta N, et al. Fas/FasL-dependent apoptosis of alveolar cells after lipopolysaccharide-induced lung injury in mice. Am J Respir Crit Care Med. 2001; 163(3 Pt 1): 762–769.
  11. Kingsley SM, Bhat BV. Role of microRNAs in sepsis. Inflamm Res. 2017; 66(7): 553–569.
  12. Adameova AD, Bhullar SK, Elimban V, et al. Activation of -adrenoceptors may not be involved in arrhythmogenesis in ischemic heart disease. Rev Cardiovasc Med. 2018; 19(3): 97–101.
  13. Yang Q, Zhang D, Li Ya, et al. Paclitaxel alleviated liver injury of septic mice by alleviating inflammatory response via microRNA-27a/TAB3/NF-κB signaling pathway. Biomed Pharmacother. 2018; 97: 1424–1433.
  14. Feng Y, Wang J, Yuan Y, et al. miR-539-5p inhibits experimental choroidal neovascularization by targeting CXCR7. FASEB J. 2018; 32(3): 1626–1639.
  15. Hu L, Liu Y, Wang B, et al. MiR-539-5p negatively regulates migration of rMSCs induced by Bushen Huoxue decoction through targeting Wnt5a. Int J Med Sci. 2019; 16(7): 998–1006.
  16. National Research Council Institute for Laboratory Animal R. Guide for the Care and Use of Laboratory Animals. Washington (DC): National Academies Press (US), 1996.
  17. Matsuda N, Hattori Y, Jesmin S, et al. Nuclear factor-kappaB decoy oligodeoxynucleotides prevent acute lung injury in mice with cecal ligation and puncture-induced sepsis. Mol Pharmacol. 2005; 67(4): 1018–1025.
  18. Ruiz S, Vardon-Bounes F, Merlet-Dupuy V, et al. Sepsis modeling in mice: ligation length is a major severity factor in cecal ligation and puncture. Intensive Care Med Exp. 2016; 4(1): 22.
  19. Morishita Y, Imai T, Yoshizawa H, et al. Delivery of microRNA-146a with polyethylenimine nanoparticles inhibits renal fibrosis in vivo. Int J Nanomedicine. 2015; 10: 3475–3488.
  20. Matute-Bello G, Downey G, Moore BB, et al. Acute Lung Injury in Animals Study Group. An official American Thoracic Society workshop report: features and measurements of experimental acute lung injury in animals. Am J Respir Cell Mol Biol. 2011; 44(5): 725–738.
  21. Hebbel RP, Vercellotti GM, Pace BS, et al. The HDAC inhibitors trichostatin A and suberoylanilide hydroxamic acid exhibit multiple modalities of benefit for the vascular pathobiology of sickle transgenic mice. Blood. 2010; 115(12): 2483–2490.
  22. Sevransky JE, Martin GS, Shanholtz C, et al. Mortality in sepsis versus non-sepsis induced acute lung injury. Crit Care. 2009; 13(5): R150.
  23. Sweeney RM, Griffiths M, McAuley D. Treatment of acute lung injury: current and emerging pharmacological therapies. Semin Respir Crit Care Med. 2013; 34(4): 487–498.
  24. Rittirsch D, Hoesel LM, Ward PA. The disconnect between animal models of sepsis and human sepsis. J Leukoc Biol. 2007; 81(1): 137–143.
  25. Chopra M, Reuben JS, Sharma AC. Acute lung injury:apoptosis and signaling mechanisms. Exp Biol Med (Maywood). 2009; 234(4): 361–371.
  26. Ganter MT, Roux J, Miyazawa B, et al. Interleukin-1beta causes acute lung injury via alphavbeta5 and alphavbeta6 integrin-dependent mechanisms. Circ Res. 2008; 102(7): 804–812.
  27. Cross LJ, Matthay MA. Biomarkers in acute lung injury: insights into the pathogenesis of acute lung injury. Crit Care Clin. 2011; 27(2): 355–377.
  28. Fielhaber JA, Carroll SF, Dydensborg AB, et al. Inhibition of mammalian target of rapamycin augments lipopolysaccharide-induced lung injury and apoptosis. J Immunol. 2012; 188(9): 4535–4542.
  29. Wang L, Ye Y, Su HB, et al. The anesthetic agent sevoflurane attenuates pulmonary acute lung injury by modulating apoptotic pathways. Braz J Med Biol Res. 2017; 50(3): e5747.
  30. Jeyaseelan S, Chu HW, Young SK, et al. Transcriptional profiling of lipopolysaccharide-induced acute lung injury. Infect Immun. 2004; 72(12): 7247–7256.
  31. Cai ZG, Zhang SM, Zhang Y, et al. MicroRNAs are dynamically regulated and play an important role in LPS-induced lung injury. Can J Physiol Pharmacol. 2012; 90(1): 37–43.
  32. Cao Y, Lyu YI, Tang J, et al. MicroRNAs: Novel regulatory molecules in acute lung injury/acute respiratory distress syndrome. Biomed Rep. 2016; 4(5): 523–527.
  33. Liu Y, Guan H, Zhang JL, et al. Acute downregulation of miR-199a attenuates sepsis-induced acute lung injury by targeting SIRT1. Am J Physiol Cell Physiol. 2018; 314(4): C449–C455.
  34. Fang Y, Gao F, Hao J, et al. microRNA-1246 mediates lipopolysaccharide-induced pulmonary endothelial cell apoptosis and acute lung injury by targeting angiotensin-converting enzyme 2. Am J Transl Res. 2017; 9(3): 1287–1296.
  35. Xie T, Liang J, Liu N, et al. MicroRNA-127 inhibits lung inflammation by targeting IgG Fcγ receptor I. J Immunol. 2012; 188(5): 2437–2444.
  36. Zeng Z, Gong H, Li Y, et al. Upregulation of miR-146a contributes to the suppression of inflammatory responses in LPS-induced acute lung injury. Exp Lung Res. 2013; 39(7): 275–282.
  37. Cinel I, Ark M, Dellinger P, et al. Involvement of Rho kinase (ROCK) in sepsis-induced acute lung injury. J Thorac Dis. 2012; 4(1): 30–39.

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