Transcriptomic study of lipopolysaccharide‑induced sepsis damage in a mouse heart model

Chen,Cunrong; Weng,Junting; Fang,Dexiang; Chen,Jianfei; Chen,Min

doi:10.3892/etm.2020.9086

Transcriptomic study of lipopolysaccharide‑induced sepsis damage in a mouse heart model

Authors:
- Cunrong Chen
- Junting Weng
- Dexiang Fang
- Jianfei Chen
- Min Chen
View Affiliations

Affiliations: Department of Critical Care Medicine, Union Hospital Affiliated to Fujian Medical University, Fuzhou, Fujian 350000, P.R. China, Department of Critical Care Medicine, Affiliated Hospital of Putian University, Putian, Fujian 351100, P.R. China
Published online on: July 31, 2020 https://doi.org/10.3892/etm.2020.9086
Pages: 3782-3790
Copyright: © Chen 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 is an emergency systemic illness caused by pathogen infection and the combined result of the underactivity and overactivity of a patient's own immune system. However, the molecular mechanism of this illness remains largely unknown. Lipopolysaccharide (LPS) was injected to establish a sepsis model, and heart tissue was used to analyze transcriptome changes in mice. LPS injection was used to develop a sepsis model, which resulted in cardiac tissue rearrangement and inflammatory response activation. An RNA‑sequencing‑based transcriptome assay using mouse heart tissue with or without LPS injection showed that 3,326 and 1,769 genes were upregulated and downregulated, respectively (>2‑fold changes; P<0.05). Furthermore, these differentially expressed genes were classified into 20 pathways, including ‘Wnt signaling pathway’, ‘VEGF signaling pathway’ and ‘TGF‑β signaling pathway’, and these altered genes were enriched in 41 Gene Ontology terms. The application of Wnt3a inhibited the activation of the LPS‑induced inflammatory response and activated Wnt signaling, as well as protecting against LPS‑mediated cardiac tissue damage in mice. In contrast, inhibition of Wnt signaling by injection of its inhibitor IWR induced plasminogen activator inhibitor‑1 expression and resulted in cardiac structure derangement, which was similar to the symptoms caused by injection of LPS, suggesting that LPS‑induced damage to heart tissue may be via inhibition of Wnt signaling. The present analyses showed that Wnt signaling serves a pivotal role in sepsis development and may improve our understanding of the molecular basis underlying sepsis.

View Figures

View References

1	Michalek SM, Moore RN, McGhee JR, Rosenstreich DL and Mergenhagen SE: The primary role of lymphoreticular cells in the mediation of host responses to bacterial endotoxim. J Infect Dis. 141:55–63. 1980.PubMed/NCBI View Article : Google Scholar
2	Beeson PB: Technical Assistance of Elizabeth Roberts: Tolerance to bacterial pyrogens: I. Factors influencing its development. J Exp Med. 86:29–38. 1947.PubMed/NCBI View Article : Google Scholar
3	Singer M, Deutschman CS, Seymour CW, Shankar-Hari M, Annane D, Bauer M, Bellomo R, Bernard GR, Chiche JD, Coopersmith CM, et al: The third international consensus definitions for sepsis and septic shock (sepsis-3). JAMA. 315:801–810. 2016.PubMed/NCBI View Article : Google Scholar
4	Seymour CW, Liu VX, Iwashyna TJ, Brunkhorst FM, Rea TD, Scherag A, Rubenfeld G, Kahn JM, Shankar-Hari M, Singer M, et al: Assessment of clinical criteria for sepsis: For the third international consensus definitions for sepsis and septic shock (sepsis-3). JAMA. 315:762–774. 2016.PubMed/NCBI View Article : Google Scholar
5	Engel C, Brunkhorst FM, Bone HG, Brunkhorst R, Gerlach H, Grond S, Gruendling M, Huhle G, Jaschinski U, John S, et al: Epidemiology of sepsis in Germany: Results from a national prospective multicenter study. Intensive Care Med. 33:606–618. 2007.PubMed/NCBI View Article : Google Scholar
6	Angus DC and Wax RS: Epidemiology of sepsis: An update. Crit Care Med. 29 (7 Suppl):S109–S116. 2001.PubMed/NCBI View Article : Google Scholar
7	Cohen J: The immunopathogenesis of sepsis. Nature. 420:885–891. 2002.PubMed/NCBI View Article : Google Scholar
8	Drosatos K, Lymperopoulos A, Kennel PJ, Pollak N, Schulze PC and Goldberg IJ: Pathophysiology of sepsis-related cardiac dysfunction: Driven by inflammation, energy mismanagement, or both? Curr Heart Fail Rep. 12:130–140. 2015.PubMed/NCBI View Article : Google Scholar
9	Harbarth S, Holeckova K, Froidevaux C, Pittet D, Ricou B, Grau GE, Vadas L and Pugin J: Geneva Sepsis Network: Diagnostic value of procalcitonin, interleukin-6, and interleukin-8 in critically ill patients admitted with suspected sepsis. Am J Respir Crit Care Med. 164:396–402. 2001.PubMed/NCBI View Article : Google Scholar
10	Simon L, Gauvin F, Amre DK, Saint-Louis P and Lacroix J: Serum procalcitonin and C-reactive protein levels as markers of bacterial infection: A systematic review and meta-analysis. Clin Infect Dis. 39:206–217. 2004.PubMed/NCBI View Article : Google Scholar
11	Bozza FA, Salluh JI, Japiassu AM, Soares M, Assis EF, Gomes RN, Bozza MT, Castro-Faria-Neto HC and Bozza PT: Cytokine profiles as markers of disease severity in sepsis: A multiplex analysis. Crit Care. 11(R49)2007.PubMed/NCBI View Article : Google Scholar
12	Vaschetto R, Nicola S, Olivieri C, Boggio E, Piccolella F, Mesturini R, Damnotti F, Colombo D, Navalesi P, Della Corte F, et al: Serum levels of osteopontin are increased in SIRS and sepsis. Intensive Care Med. 34:2176–2184. 2008.PubMed/NCBI View Article : Google Scholar
13	Brenner T, Rosenhagen C, Steppan J, Lichtenstern C, Weitz J, Bruckner T, Martin EO, Hoffmann U, Weigand MA and Hofer S: Redox responses in patients with sepsis: High correlation of thioredoxin-1 and macrophage migration inhibitory factor plasma levels. Mediators Inflamm. 2010(985614)2010.PubMed/NCBI View Article : Google Scholar
14	Bae JS: Role of high mobility group box 1 in inflammatory disease: Focus on sepsis. Arch Pharm Res. 35:1511–1523. 2012.PubMed/NCBI View Article : Google Scholar
15	Wu Y, Wang F, Fan X, Bao R, Bo L, Li J and Deng X: Accuracy of plasma sTREM-1 for sepsis diagnosis in systemic inflammatory patients: A systematic review and meta-analysis. Crit Care. 16(R229)2012.PubMed/NCBI View Article : Google Scholar
16	Backes Y, van der Sluijs KF, Mackie DP, Tacke F, Koch A, Tenhunen JJ and Schultz MJ: Usefulness of suPAR as a biological marker in patients with systemic inflammation or infection: A systematic review. Intensive Care Med. 38:1418–1428. 2012.PubMed/NCBI View Article : Google Scholar
17	Rautanen A, Mills TC, Gordon AC, Hutton P, Steffens M, Nuamah R, Chiche JD, Parks T, Chapman SJ, Davenport EE, et al: Genome-wide association study of survival from sepsis due to pneumonia: An observational cohort study. Lancet Respir Med. 3:53–60. 2015.PubMed/NCBI View Article : Google Scholar
18	Scherag A, Schöneweck F, Kesselmeier M, Taudien S, Platzer M, Felder M, Sponholz C, Rautanen A, Hill AVS, Hinds CJ, et al: Genetic factors of the disease course after sepsis: A genome-wide study for 28 day mortality. EbioMedicine. 12:239–246. 2016.PubMed/NCBI View Article : Google Scholar
19	Wang L, Ko ER2, Gilchrist JJ, Pittman KJ, Rautanen A, Pirinen M, Thompson JW, Dubois LG, Langley RJ, Jaslow SL, et al: Human genetic and metabolite variation reveals that methylthioadenosine is a prognostic biomarker and an inflammatory regulator in sepsis. Sci Adv. 3(e1602096)2017.PubMed/NCBI View Article : Google Scholar
20	Brown MA and Jones WK: NF-kappaB action in sepsis: The innate immune system and the heart. Front Biosci. 9:1201–1217. 2004.PubMed/NCBI View Article : Google Scholar
21	Li Y, Ge S, Peng Y and Chen X: Inflammation and cardiac dysfunction during sepsis, muscular dystrophy, and myocarditis. Burns Trauma. 1:109–121. 2013.PubMed/NCBI View Article : Google Scholar
22	MacDonald BT, Tamai K and He X: Wnt/beta-catenin signaling: Components, mechanisms, and diseases. Dev Cell. 17:9–26. 2009.PubMed/NCBI View Article : Google Scholar
23	Gordon MD and Nusse R: Wnt signaling: Multiple pathways, multiple receptors, and multiple transcription factors. J Biol Chem. 281:22429–22433. 2006.PubMed/NCBI View Article : Google Scholar
24	Shtutman M, Zhurinsky J, Simcha I, Albanese C, D'Amico M, Pestell R and Ben-Ze'ev A: The cyclin D1 gene is a target of the beta-catenin/LEF-1 pathway. Proc Natl Acad Sci USA. 96:5522–5527. 1999.PubMed/NCBI View Article : Google Scholar
25	Moon RT, Kohn AD, De Ferrari GV and Kaykas A: WNT and β-catenin signalling: Diseases and therapies. Nat Rev Genet. 5:691–701. 2004.PubMed/NCBI View Article : Google Scholar
26	Clevers H: Wnt/β-catenin signaling in development and disease. Cell. 127:469–480. 2006.PubMed/NCBI View Article : Google Scholar
27	Brack AS, Conboy MJ, Roy S, Lee M, Kuo CJ, Keller C and Rando TA: Increased Wnt signaling during aging alters muscle stem cell fate and increases fibrosis. Science. 317:807–810. 2007.PubMed/NCBI View Article : Google Scholar
28	Aberle H, Bauer A, Stappert J, Kispert A and Kemler R: Beta-catenin is a target for the ubiquitin-proteasome pathway. EMBO J. 16:3797–3804. 1997.PubMed/NCBI View Article : Google Scholar
29	National Research Council: Guide for the care and use of laboratory animals. National Academies Press, Washington, DC, 1996.
30	Leary S: AVMA Guidelines for the Euthanasia of Animals: 2013 Edition. American Veterinary Medical Association. J Am Veterinary Med Association, 2013.
31	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
32	Rani B: RNA Sequencing (RNA-Seq): Method and Applications. Int J Pure App Biosci. 6:167–173. 2018.
33	Kim D, Pertea G, Trapnell C, Pimentel H, Kelley R and Salzberg SL: TopHat2: Accurate alignment of transcriptomes in the presence of insertions, deletions and gene fusions. Genome Biol. 14(R36)2013.PubMed/NCBI View Article : Google Scholar
34	Langmead B and Salzberg SL: Fast gapped-read alignment with Bowtie 2. Nat Methods. 9:357–359. 2012.PubMed/NCBI View Article : Google Scholar
35	Gene Ontology Consortium. The gene ontology (GO) project in 2006. Nucleic Acids Res 34: D322-D326, 2006.
36	Dupuy D, Bertin N, Hidalgo CA, Venkatesan K, Tu D, Lee D, Rosenberg J, Svrzikapa N, Blanc A, Carnec A, et al: Genome-scale analysis of in vivo spatiotemporal promoter activity in Caenorhabditis elegans. Nat Biotechnol. 25:663–668. 2007.PubMed/NCBI View Article : Google Scholar
37	Dunnick JK, Brix A, Cunny H, Vallant M and Shockley KR: Characterization of polybrominated diphenyl ether toxicity in Wistar Han rats and use of liver microarray data for predicting disease susceptibilities. Toxicol Pathol. 40:93–106. 2012.PubMed/NCBI View Article : Google Scholar
38	Kanehisa M, Goto S, Kawashima S and Nakaya A: The KEGG databases at Genome Net Nucleic Acids. Res. 30:42–46. 2002.PubMed/NCBI View Article : Google Scholar
39	Nishimura D: BioCarta. Biotech Software Internet Rep. 2:117–120. 200.
40	Matthews L, Gopinath G, Gillespie M, Caudy M, Croft D, de Bono B, Garapati P, Hemish J, Hermjakob H, Jassal B, et al: Reactome knowledgebase of human biological pathways and processes. Nucleic Acids Res. 37 (Database Issue):D619–D622. 2009.PubMed/NCBI View Article : Google Scholar
41	Draghici S, Khatri P, Tarca AL, Amin K, Done A, Voichita C, Georgescu C and Romero R: A systems biology approach for pathway level analysis. Genome Res. 17:1537–1545. 2007.PubMed/NCBI View Article : Google Scholar
42	Turner A, Tsamitros M and Bellomo R: Myocardial cell injury in septic shock. Crit Care Med. 27:2035–2036. 1999.PubMed/NCBI View Article : Google Scholar
43	Hochstadt A, Meroz Y and Landesberg G: Myocardial dysfunction in severe sepsis and septic shock: More questions than answers? J Cardiothorac Vasc Anesth. 25:526–535. 2011.PubMed/NCBI View Article : Google Scholar
44	Dios S, Balseiro P, Costa MM, Romero A, Boltaña S, Roher N, Mackenzie S, Figueras A and Novoa B: The involvement of cholesterol in sepsis and tolerance to lipopolysaccharide highlighted by the transcriptome analysis of zebrafish (Danio rerio). Zebrafis. 11:421–433. 2014.PubMed/NCBI View Article : Google Scholar
45	Traenckner EB, Pahl HL, Henkel T, Schmidt KN, Wilk S and Baeuerle PA: Phosphorylation of human I kappa B-alpha on serines 32 and 36 controls I kappa B-alpha proteolysis and NF-kappa B activation in response to diverse stimuli. EMBO J. 14:2876–2883. 1995.PubMed/NCBI
46	Jaffer U, Wade RG and Gourlay T: Cytokines in the systemic inflammatory response syndrome: A review. HSR Proc Intensive Care Cardiovasc Anesth. 2:161–175. 2010.PubMed/NCBI
47	Carmeliet P: VEGF as a key mediator of angiogenesis in cancer. Oncology. 69:4–10. 2005.PubMed/NCBI View Article : Google Scholar
48	Derynck R, Akhurst RJ and Balmain A: TGF-β signaling in tumor suppression and cancer progression. Nature Genet. 29:117–129. 2001.PubMed/NCBI View Article : Google Scholar
49	Planavila A, Redondo-Angulo I, Ribas F, Garrabou G, Casademont J, Giralt M and Villarroya F: Fibroblast growth factor 21 protects the heart from oxidative stress. Cardiovasc Res. 106:19–31. 2015.PubMed/NCBI View Article : Google Scholar
50	Singhal G, Fisher FM, Chee MJ, Tan TG El Ouaamari A, Adams AC, Najarian R, Kulkarni RN, Benoist C, Flier JS and Maratos-Flier E: Fibroblast Growth Factor 21 (FGF21) protects against high fat diet induced inflammation and islet hyperplasia in pancreas. PLoS One. 11(e0148252)2016.PubMed/NCBI View Article : Google Scholar