MicroRNA‑139‑5p improves sepsis‑induced lung injury by targeting Rho‑kinase1

Zhao,Xinmin; Wang,Mengmeng; Sun,Zhaorui; Zhuang,Suyuan; Zhang,Wei; Yang,Zhizhou; Han,Xiaoqin; Nie,Shinan

doi:10.3892/etm.2021.10493

MicroRNA‑139‑5p improves sepsis‑induced lung injury by targeting Rho‑kinase1

Authors:
- Xinmin Zhao
- Mengmeng Wang
- Zhaorui Sun
- Suyuan Zhuang
- Wei Zhang
- Zhizhou Yang
- Xiaoqin Han
- Shinan Nie
View Affiliations

Affiliations: Department of Emergency Medicine, Jinling Hospital, The First School of Clinical Medicine, Southern Medical University, Nanjing, Jiangsu 210002, P.R. China, Department of Emergency Medicine, Jinling Hospital, Medical School of Nanjing University, Nanjing, Jiangsu 210002, P.R. China
Published online on: July 26, 2021 https://doi.org/10.3892/etm.2021.10493
Article Number: 1059
Copyright: © Zhao et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Sepsis‑induced acute lung injury (ALI) is an inflammatory process that involves inflammatory cytokine production and cell apoptosis. In the present study, the regulatory role of microRNA (miR)‑139‑5p in sepsis‑induced ALI was investigated using a murine model of cecal ligation puncture (CLP) and an in vitro model using lipopolysaccharide (LPS)‑induced normal human bronchial epithelial cells (NHBEs). Sepsis‑induced pathological changes in the lungs of ALI mice were detected using hematoxylin and eosin staining. Lung water content was determined, and the expression of proinflammatory cytokines in the bronchoalveolar lavage fluid and serum of sepsis‑induced ALI mice were quantified using ELISA. The levels of oxidative stress in lung tissues were determined using commercial kits. The degree of apoptosis was determined using a TUNEL assay. The expression levels of miR‑139‑5p and Rho‑kinase 1 (ROCK1) were determined using reverse transcription‑quantitative PCR and western blot analyses. A dual‑luciferase reporter assay was used to confirm the direct targeting of ROCK1 by miR‑139‑5p. NHBEs were co‑transfected with vectors expressing ROCK1 (or empty vector) and miR‑139‑5p mimics or control mimics prior to LPS treatment. The transcriptional activity of caspase‑3, the ratio of apoptotic cells, the expression levels of mucin 5AC, mucin 1, TNF‑α, IL‑1β, IL‑6, NLR family pyrin domain containing 3, apoptosis‑associated speck‑like protein containing a CARD and caspase‑1 were evaluated. Compared with the normal group, mice that underwent CLP exhibited abnormal lung morphology, enhanced production of TNF‑α, IL‑1β and IL‑6, increased reactive oxygen species (ROS), malondialdehyde and lactate dehydrogenase levels, an increased proportion of apoptotic cells and increased ROCK1 expression. Superoxide dismutase, glutathione peroxidase and miR‑139‑5p levels were decreased following CLP. In the NHBEs, stimulation with LPS caused a marked increase in inflammatory cytokine levels and apoptosis compared with the untreated cells. Overexpression of miR‑139‑5p attenuated cell apoptosis and inflammation. Overexpression of ROCK1 in NHBEs restored the ROS levels and proinflammatory cytokine production inhibited by miR‑139‑5p. In conclusion, miR‑139‑5p alleviated sepsis‑induced ALI via suppression of its downstream target, ROCK1, suggesting that miR‑139‑5p may hold promise in the treatment of sepsis‑induced ALI.

View Figures

View References

1	Cecconi M, Evans L, Levy M and Rhodes A: Sepsis and septic shock. Lancet. 392:75–87. 2018.PubMed/NCBI View Article : Google Scholar
2	Vincent JL, Marshall JC, Namendys-Silva SA, François B, Martin-Loeches I, Lipman J, Reinhart K, Antonelli M, Pickkers P, Njimi H, et al: Assessment of the worldwide burden of critical illness: The intensive care over nations (ICON) audit. Lancet Respir Med. 2:380–386. 2014.PubMed/NCBI View Article : Google Scholar
3	Singer BH, Dickson RP, Denstaedt SJ, Newstead MW, Kim K, Falkowski NR, Erb-Downward JR, Schmidt TM, Huffnagle GB and Standiford TJ: Bacterial dissemination to the brain in Sepsis. Am J Respir Crit Care Med. 197:747–756. 2018.PubMed/NCBI View Article : Google Scholar
4	Merx MW and Weber C: Sepsis and the heart. Circulation. 116:793–802. 2007.PubMed/NCBI View Article : Google Scholar
5	Meneses G, Cardenas G, Espinosa A, Rassy D, Pérez-Osorio IN, Bárcena B, Fleury A, Besedovsky H, Fragoso G and Sciutto E: Sepsis: Developing new alternatives to reduce neuroinflammation and attenuate brain injury. Ann N Y Acad Sci. 1437:43–56. 2019.PubMed/NCBI View Article : Google Scholar
6	Hato T, Maier B, Syed F, Myslinski J, Zollman A, Plotkin Z, Eadon MT and Dagher PC: Bacterial sepsis triggers an antiviral response that causes translation shutdown. J Clin Invest. 129:296–309. 2019.PubMed/NCBI View Article : Google Scholar
7	Fowler AA III, Truwit JD, Hite RD, Morris PE, DeWilde C, Priday A, Fisher B, Thacker LR II, Natarajan R, Brophy DF, et al: Effect of vitamin C infusion on organ failure and biomarkers of inflammation and vascular injury in patients with sepsis and severe acute respiratory failure: The CITRIS-ALI randomized clinical trial. JAMA. 322:1261–1270. 2019.PubMed/NCBI View Article : Google Scholar
8	Fernandez FG, Kosinski AS, Furnary AP, Onaitis M, Kim S, Habib RH, Tong BC, Cowper P, Boffa D, Jacobs JP, et al: Differential effects of operative complications on survival after surgery for primary lung cancer. J Thorac Cardiovasc Surg. 155:1254–1264.e1. 2018.PubMed/NCBI View Article : Google Scholar
9	Vazquez-Medina JP, Tao JQ, Patel P, Bannitz-Fernandes R, Dodia C, Sorokina EM, Feinstein SI, Chatterjee S and Fisher AB: Genetic inactivation of the phospholipase A2 activity of peroxiredoxin 6 in mice protects against LPS-induced acute lung injury. Am J Physiol Lung Cell Mol Physiol. 316:L656–L668. 2019.PubMed/NCBI View Article : Google Scholar
10	Guo RF and Ward PA: Role of oxidants in lung injury during sepsis. Antioxid Redox Signal. 9:1991–2002. 2007.PubMed/NCBI View Article : Google Scholar
11	Yi L, Chang M, Zhao Q, Zhou Z, Huang X, Guo F and Huan J: Genistein-3'-sodium sulphonate protects against lipopolysaccharide-induced lung vascular endothelial cell apoptosis and acute lung injury via BCL-2 signalling. J Cell Mol Med. 24:1022–1035. 2020.PubMed/NCBI View Article : Google Scholar
12	Swisher EM, Lin KK, Oza AM, Scott CL, Giordano H, Sun J, Konecny GE, Coleman RL, Tinker AV, O'Malley DM, et al: Rucaparib in relapsed, platinum-sensitive high-grade ovarian carcinoma (ARIEL2 Part 1): An international, multicentre, open-label, phase 2 trial. Lancet Oncol. 18:75–87. 2017.PubMed/NCBI View Article : Google Scholar
13	Hennessy EJ, Parker AE and O'Neill LA: Targeting Toll-like receptors: Emerging therapeutics? Nat Rev Drug Discov. 9:293–307. 2010.PubMed/NCBI View Article : Google Scholar
14	Seeley JJ, Baker RG, Mohamed G, Bruns T, Hayden MS, Deshmukh SD, Freedberg DE and Ghosh S: Induction of innate immune memory via microRNA targeting of chromatin remodelling factors. Nature. 559:114–119. 2018.PubMed/NCBI View Article : Google Scholar
15	Gotts JE and Matthay MA: Sepsis: Pathophysiology and clinical management. BMJ. 353(i1585)2016.PubMed/NCBI View Article : Google Scholar
16	Zhu M, Zhang W, Ma J, Dai Y, Zhang Q, Liu Q, Yang B and Li G: MicroRNA-139-5p regulates chronic inflammation by suppressing nuclear factor-κB activity to inhibit cell proliferation and invasion in colorectal cancer. Exp Ther Med. 18:4049–4057. 2019.PubMed/NCBI View Article : Google Scholar
17	Qu Y, Wu J, Chen D, Zhao F, Liu J, Yang C, Wei D, Ferriero DM and Mu D: MiR-139-5p inhibits HGTD-P and regulates neuronal apoptosis induced by hypoxia-ischemia in neonatal rats. Neurobiol Dis. 63:184–193. 2014.PubMed/NCBI View Article : Google Scholar
18	Huang N, Guo W, Ren K, Li W, Jiang Y, Sun J, Dai W and Zhao W: LncRNA AFAP1-AS1 Supresses miR-139-5p and promotes cell proliferation and chemotherapy resistance of non-small cell lung cancer by competitively upregulating RRM2. Front Oncol. 9(1103)2019.PubMed/NCBI View Article : Google Scholar
19	Wang Y, Wang X, Liu W and Zhang L: Role of the Rho/ROCK signaling pathway in the protective effects of fasudil against acute lung injury in septic rats. Mol Med Rep. 18:4486–4498. 2018.PubMed/NCBI View Article : Google Scholar
20	Ruiz S, Vardon-Bounes F, Merlet-Dupuy V, Conil JM, Buléon M, Fourcade O, Tack I and Minville V: Sepsis modeling in mice: Ligation length is a major severity factor in cecal ligation and puncture. Intensive Care Med Exp. 4(22)2016.PubMed/NCBI View Article : Google Scholar
21	Zhuo Y, Li D, Cui L, Li C, Zhang S, Zhang Q, Zhang L, Wang X and Yang L: Treatment with 3,4-dihydroxyphenylethyl alcohol glycoside ameliorates sepsis-induced ALI in mice by reducing inflammation and regulating M1 polarization. Biomed Pharmacother. 116(109012)2019.PubMed/NCBI View Article : Google Scholar
22	Matute-Bello G, Downey G, Moore BB, Groshong SD, Matthay MA, Slutsky AS and Kuebler WM: An official American Thoracic Society workshop report: Features and measurements of experimental acute lung injury in animals. Am J Respir Cell Mol Biol. 44:725–738. 2011.PubMed/NCBI View Article : Google Scholar
23	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods. 25:402–408. 2001.PubMed/NCBI View Article : Google Scholar
24	Shang G, Jin Y, Zheng Q, Shen X, Yang M, Li Y and Zhang L: Histology and oncogenic driver alterations of lung adenocarcinoma in Chinese. Am J Cancer Res. 9:1212–1223. 2019.PubMed/NCBI
25	Englert JA, Bobba C and Baron RM: Integrating molecular pathogenesis and clinical translation in sepsis-induced acute respiratory distress syndrome. JCI Insight. 4(e124061)2019.PubMed/NCBI View Article : Google Scholar
26	Konduri K, Gallant JN, Chae YK, Giles FJ, Gitlitz BJ, Gowen K, Ichihara E, Owonikoko TK, Peddareddigari V, Ramalingam SS, et al: EGFR fusions as novel therapeutic targets in lung cancer. Cancer Discov. 6:601–611. 2016.PubMed/NCBI View Article : Google Scholar
27	Dejager L, Pinheiro I, Dejonckheere E and Libert C: Cecal ligation and puncture: The gold standard model for polymicrobial sepsis? Trends Microbiol. 19:198–208. 2011.PubMed/NCBI View Article : Google Scholar
28	Hong HC, Chuang CH, Huang WC, Weng SL, Chen CH, Chang KH, Liao KW and Huang HD: A panel of eight microRNAs is a good predictive parameter for triple-negative breast cancer relapse. Theranostics. 10:8771–8789. 2020.PubMed/NCBI View Article : Google Scholar
29	Du F, Cao T, Xie H, Li T, Sun L, Liu H, Guo H, Wang X, Liu Q, Kim T, et al: KRAS Mutation-Responsive miR-139-5p inhibits colorectal cancer progression and is repressed by Wnt Signaling. Theranostics. 10:7335–7350. 2020.PubMed/NCBI View Article : Google Scholar
30	Xiu D, Liu L, Cheng M, Sun X and Ma X: Knockdown of lncRNA TUG1 enhances radiosensitivity of prostate cancer via the TUG1/miR-139-5p/SMC1A Axis. Onco Targets Ther. 13:2319–2331. 2020.PubMed/NCBI View Article : Google Scholar
31	Wen J, Wang G, Xie X, Lin G, Yang H, Luo K, Liu Q, Ling Y, Xie X, Lin P, et al: Prognostic value of a Four-miRNA signature in patients with lymph node positive locoregional esophageal squamous cell carcinoma undergoing complete surgical resection. Ann Surg. 273:523–531. 2021.PubMed/NCBI View Article : Google Scholar
32	Pajic M, Froio D, Daly S, Doculara L, Millar E, Graham PH, Drury A, Steinmann A, de Bock CE, Boulghourjian A, et al: MiR-139-5p modulates radiotherapy resistance in breast cancer by repressing multiple gene networks of DNA repair and ROS defense. Cancer Res. 78:501–515. 2018.PubMed/NCBI View Article : Google Scholar
33	Zou F, Mao R, Yang L, Lin S, Lei K, Zheng Y, Ding Y, Zhang P, Cai G, Liang X and Liu J: Targeted deletion of miR-139-5p activates MAPK, NF-κB and STAT3 signaling and promotes intestinal inflammation and colorectal cancer. FEBS J. 283:1438–1452. 2016.PubMed/NCBI View Article : Google Scholar
34	Li C, Ma D, Yang J, Lin X and Chen B: MiR-202-5p inhibits the migration and invasion of osteosarcoma cells by targeting ROCK1. Oncol Lett. 16:829–834. 2018.PubMed/NCBI View Article : Google Scholar
35	Roberto GM, Delsin LEA, Vieira GM, Silva MO, Hakime RG, Gava NF, Engel EE, Scrideli CA, Tone LG and Brassesco MS: ROCK1-PredictedmicroRNAs dysregulation contributes to tumor progression in ewing sarcoma. Pathol Oncol Res. 26:133–139. 2020.PubMed/NCBI View Article : Google Scholar
36	Tian K, Sun D, Chen M, Yang Y, Wang F, Guo T and Shi Z: Long Noncoding RNA X-Inactive specific transcript facilitates cellular functions in melanoma via miR-139-5p/ROCK1 Pathway. Onco Targets Ther. 13:1277–1287. 2020.PubMed/NCBI View Article : Google Scholar