MiR-539-5p alleviates sepsis-induced acute lung injury by targeting ROCK1
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.
Keywords: mouseacute lung injurysepsisinflammationMPVEC cellsmiRNAROCK1caspase-3cytokines
References
- Sagy M, Al-Qaqaa Y, Kim P. Definitions and pathophysiology of sepsis. Curr Probl Pediatr Adolesc Health Care. 2013; 43(10): 260–263.
- Gando S. Microvascular thrombosis and multiple organ dysfunction syndrome. Crit Care Med. 2010; 38(2 Suppl): S35–S42.
- 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.
- Johnson ER, Matthay MA. Acute lung injury: epidemiology, pathogenesis, and treatment. J Aerosol Med Pulm Drug Deliv. 2010; 23(4): 243–252.
- 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.
- 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.
- 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.
- Martin TR, Nakamura M, Matute-Bello G. The role of apoptosis in acute lung injury. Crit Care Med. 2003; 31(4 Suppl): S184–S188.
- 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.
- 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.
- Kingsley SM, Bhat BV. Role of microRNAs in sepsis. Inflamm Res. 2017; 66(7): 553–569.
- 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.
- 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.
- 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.
- 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.
- National Research Council Institute for Laboratory Animal R. Guide for the Care and Use of Laboratory Animals. Washington (DC): National Academies Press (US), 1996.
- 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.
- 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.
- 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.
- 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.
- 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.
- Sevransky JE, Martin GS, Shanholtz C, et al. Mortality in sepsis versus non-sepsis induced acute lung injury. Crit Care. 2009; 13(5): R150.
- 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.
- Rittirsch D, Hoesel LM, Ward PA. The disconnect between animal models of sepsis and human sepsis. J Leukoc Biol. 2007; 81(1): 137–143.
- Chopra M, Reuben JS, Sharma AC. Acute lung injury:apoptosis and signaling mechanisms. Exp Biol Med (Maywood). 2009; 234(4): 361–371.
- 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.
- 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.
- 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.
- 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.
- Jeyaseelan S, Chu HW, Young SK, et al. Transcriptional profiling of lipopolysaccharide-induced acute lung injury. Infect Immun. 2004; 72(12): 7247–7256.
- 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.
- 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.
- 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.
- 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.
- 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.
- 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.
- 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.
