Wnt3a suppresses Pseudomonas aeruginosa‑induced inflammation and promotes bacterial killing in macrophages

Chen,Kang; Fu,Qiang; Li,Dandan; Wu,Yongjian; Sun,Shijun; Zhang,Xiuming

doi:10.3892/mmr.2016.4869

Wnt3a suppresses Pseudomonas aeruginosa‑induced inflammation and promotes bacterial killing in macrophages

Authors:
- Kang Chen
- Qiang Fu
- Dandan Li
- Yongjian Wu
- Shijun Sun
- Xiuming Zhang
View Affiliations

Affiliations: Division of Clinical Laboratory, Zhongshan Hospital of Sun Yat‑Sen University, Zhongshan, Guangdong 528403, P.R. China, Department of Immunology, Institute of Human Virology, Zhongshan School of Medicine, Sun Yat‑Sen University, Guangzhou, Guangdong 510080, P.R. China
Published online on: February 5, 2016 https://doi.org/10.3892/mmr.2016.4869
Pages: 2439-2446
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

Pseudomonas aeruginosa (PA) is a common Gram‑negative bacterium and can cause serious infections, including hospital‑acquired pneumonia, suppurative bacterial keratitis and acute burn wound infection. The pathogenesis of PA infections is closely associated with excessive inflammatory responses and bacterial virulence factors. Wingless‑type MMTV integration site family, member 3A (Wnt3a), an upstream mediator in the canonical Wnt signaling pathway, has been implicated as a regulator of inflammation. However, its role in PA‑induced inflammation and bacterial clearance remains unknown. In the present study, the efficacy of Wnt3a conditioned media (Wnt3a‑CM) was assessed using western blotting and immunofluorescence, which showed that β‑catenin, a downstream molecule of Wnt3a, was upregulated and translocated to the nucleus following exposure to 50% Wnt3a‑CM for 6 h. To explore the role of Wnt3a in PA‑induced inflammation, the mRNA levels of pro‑inflammatory cytokines and apoptosis in macrophages were measured using reverse transcription‑quantitative polymerase chain reaction and flow cytometry, respectively. This indicated that Wnt3a suppressed inflammation by reducing the production of pro‑inflammatory cytokines and by promoting apoptosis in macrophages. Furthermore, the mechanism of macrophage‑mediated bacterial killing was investigated, and the results indicated that Wnt3a enhanced macrophage‑mediated intracellular bacterial killing via the induction of the production of cathelicidin‑related antimicrobial peptide and β‑defensins 1. Taken together, the current study explored the role of Wnt3a in inflammation and bacterial invasion, which may provide an improved understanding of host resistance to PA infection.

View Figures

View References

1	Lovewell RR, Patankar YR and Berwin B: Mechanisms of phagocytosis and host clearance of Pseudomonas aeruginosa. Am J Physiol Lung Cell Mol Physiol. 306:L591–L603. 2014. View Article : Google Scholar : PubMed/NCBI
2	Hazlett LD: Corneal response to Pseudomonas aeruginosa infection. Prog Retin Eye Res. 23:1–30. 2004. View Article : Google Scholar : PubMed/NCBI
3	Bielecki P, Glik J, Kawecki M and Martins dos Santos VA: Towards understanding Pseudomonas aeruginosa burn wound infections by profiling gene expression. Biotechnol Lett. 30:777–790. 2008. View Article : Google Scholar
4	Kowalski RP, Romanowski EG, Mah FS, Shanks RM and Gordon YJ: Topical levofloxacin 1.5% overcomes in vitro resistance in rabbit keratitis models. Acta Ophthalmol. 88:e120–e125. 2010. View Article : Google Scholar : PubMed/NCBI
5	McCormick C, Caballero A, Tang A, Balzli C, Song J and O'Callaghan R: Effectiveness of a new tobramycin (0.3%) and dexamethasone (0.05%) formulation in the treatment of experimental Pseudomonas keratitis. Curr Med Res Opin. 24:1569–1575. 2008. View Article : Google Scholar : PubMed/NCBI
6	Mohammadpour M, Mohajernezhadfard Z, Khodabande A and Vahedi P: Antibiotic susceptibility patterns of Pseudomonas corneal ulcers in contact lens wearers. Middle East Afr J Ophthalmol. 18:228–231. 2011. View Article : Google Scholar : PubMed/NCBI
7	Hazlett LD: Pathogenic mechanisms of P. aeruginosa keratitis: A review of the role of T cells, Langerhans cells, PMN and cytokines. DNA Cell Biol. 21:383–390. 2002. View Article : Google Scholar : PubMed/NCBI
8	Kernacki KA, Goebel DJ, Poosch MS and Hazlett LD: Early cytokine and chemokine gene expression during Pseudomonas aeruginosa corneal infection in mice. Infect Immun. 66:376–379. 1998.PubMed/NCBI
9	Zhou T, Hu Y, Chen Y, Zhou KK, Zhang B, Gao G and Ma JX: The pathogenic role of the canonical Wnt pathway in age-related macular degeneration. Invest Ophthalmol Vis Sci. 51:4371–4379. 2010. View Article : Google Scholar :
10	You J, Nguyen AV, Albers CG, Lin F and Holcombe RF: Wnt pathway-related gene expression in inflammatory bowel disease. Dig Dis Sci. 53:1013–1019. 2008. View Article : Google Scholar
11	Miao CG, Yang YY, He X, Li XF, Huang C, Huang Y, Zhang L, Lv XW, Jin Y and Li J: Wnt signaling pathway in rheumatoid arthritis, with special emphasis on the different roles in synovial inflammation and bone remodeling. Cell Signal. 25:2069–2078. 2013. View Article : Google Scholar : PubMed/NCBI
12	Wodarz A and Nusse R: Mechanisms of Wnt signaling in development. Annu Rev Cell Dev Biol. 14:59–88. 1998. View Article : Google Scholar
13	Aumiller V, Balsara N, Wilhelm J, Günther A and Königshoff M: WNT/β-catenin signaling induces IL-1β expression by alveolar epithelial cells in pulmonary fibrosis. Am J Respir Cell Mol Biol. 49:96–104. 2013. View Article : Google Scholar : PubMed/NCBI
14	Liu X, Lu R, Wu S, Zhang YG, Xia Y, Sartor RB and Sun J: Wnt2 inhibits enteric bacterial-induced inflammation in intestinal epithelial cells. Inflamm Bowel Dis. 18:418–429. 2012. View Article : Google Scholar :
15	Liu X, Wu S, Xia Y, Li XE, Xia Y, Zhou ZD and Sun J: Wingless homolog Wnt11 suppresses bacterial invasion and inflammation in intestinal epithelial cells. Am J Physiol Gastrointest Liver Physiol. 301:G992–G1003. 2011. View Article : Google Scholar : PubMed/NCBI
16	Zimmerman ZF, Kulikauskas RM, Bomsztyk K, Moon RT and Chien AJ: Activation of Wnt/β-catenin signaling increases apoptosis in melanoma cells treated with trail. PLoS One. 8:e695932013. View Article : Google Scholar
17	Gui S, Yuan G, Wang L, Zhou L, Xue Y, Yu Y, Zhang J, Zhang M, Yang Y and Wang DW: Wnt3a regulates proliferation, apoptosis and function of pancreatic NIT-1 beta cells via activation of IRS2/PI3K signaling. J Cell Biochem. 114:1488–1497. 2013. View Article : Google Scholar : PubMed/NCBI
18	Kim JS, Yeo S, Shin DG, Bae YS, Lee JJ, Chin BR, Lee CH and Baek SH: Glycogen synthase kinase 3beta and beta-catenin pathway is involved in toll-like receptor 4-mediated NADPH oxidase 1 expression in macrophages. FEBS J. 277:2830–2837. 2010. View Article : Google Scholar : PubMed/NCBI
19	Du Q, Park KS, Guo Z, He P, Nagashima M, Shao L, Sahai R, Geller DA and Hussain SP: Regulation of human nitric oxide synthase 2 expression by Wnt beta-catenin signaling. Cancer Res. 66:7024–7031. 2006. View Article : Google Scholar : PubMed/NCBI
20	Huang LC, Reins RY, Gallo RL and McDermott AM: Cathelicidin-deficient (Cnlp −/−) mice show increased susceptibility to Pseudomonas aeruginosa keratitis. Invest Ophthalmol Vis Sci. 48:4498–4508. 2007. View Article : Google Scholar : PubMed/NCBI
21	Kumar A, Hazlett LD and Yu FS: Flagellin suppresses the inflammatory response and enhances bacterial clearance in a murine model of Pseudomonas aeruginosa keratitis. Infect Immun. 76:89–96. 2008. View Article : Google Scholar :
22	Hazlett L and Wu M: Defensins in innate immunity. Cell Tissue Res. 343:175–188. 2011. View Article : Google Scholar
23	Chen K, Yin L, Nie X, Deng Q, Wu Y, Zhu M, Li D, Li M, Wu M and Huang X: β-Catenin promotes host resistance against Pseudomonas aeruginosa keratitis. J Infect. 67:584–594. 2013. View Article : Google Scholar : PubMed/NCBI
24	Mariencheck WI, Savov J, Dong Q, Tino MJ and Wright JR: Surfactant protein A enhances alveolar macrophage phagocytosis of a live, mucoid strain of P. aeruginosa. Am J Physiol. 277:L777–L786. 1999.PubMed/NCBI
25	Neumann J, Schaale K, Farhat K, Endermann T, Ulmer AJ, Ehlers S and Reiling N: Frizzled1 is a marker of inflammatory macrophages and its ligand Wnt3a is involved in reprogramming Mycobacterium tuberculosis-infected macrophages. FASEB J. 24:4599–4612. 2010. View Article : Google Scholar : PubMed/NCBI
26	Sun J, Hobert ME, Rao AS, Neish AS and Madara JL: Bacterial activation of beta-catenin signaling in human epithelia. Am J Physiol Gastrointest Liver Physiol. 287:G220–G227. 2004. View Article : Google Scholar : PubMed/NCBI
27	Sun J, Hobert ME, Duan Y, Rao AS, He TC, Chang EB and Madara JL: Crosstalk between NF-kappaB and beta-catenin pathways in bacterial-colonized intestinal epithelial cells. Am J Physiol Gastrointest Liver Physiol. 289:G129–G137. 2005. View Article : Google Scholar : PubMed/NCBI
28	Wu X, Deng G, Hao X, Li Y, Zeng J, Ma C, He Y, Liu X and Wang Y: A caspase-dependent pathway is involved in Wnt/β-catenin signaling promoted apoptosis in Bacillus Calmette-Guerin infected RAW264.7 macrophages. Int J Mol Sci. 15:5045–5062. 2014. View Article : Google Scholar : PubMed/NCBI
29	Xu S and Gotlieb AI: Wnt3a/β-catenin increases proliferation in heart valve interstitial cells. Cardiovasc Pathol. 22:156–166. 2013. View Article : Google Scholar
30	Thomas EL, Lehrer RI and Rest RF: Human neutrophil antimicrobial activity. Rev Infect Dis. 10(Suppl 2): S450–S456. 1988. View Article : Google Scholar : PubMed/NCBI
31	Wiesner J and Vilcinskas A: Antimicrobial peptides: The ancient arm of the human immune system. Virulence. 1:440–464. 2010. View Article : Google Scholar : PubMed/NCBI