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BMC Ophthalmology
Published in: BMC Ophthalmology 1/2016

Open Access 01-12-2016 | Research article

Angiogenin ameliorates corneal opacity and neovascularization via regulating immune response in corneal fibroblasts

Authors: Seung Hoon Lee, Kyoung Woo Kim, Kwangsic Joo, Jae Chan Kim

Published in: BMC Ophthalmology | Issue 1/2016

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Abstract

Background

Angiogenin (ANG), a component of tears, is involved in the innate immune system and is related with inflammatory disease. We investigated whether ANG has an immune modulatory function in human corneal fibroblasts (HCFs).

Methods

HCFs were cultured from excised corneal tissues. The gene or protein expression levels of interleukin (IL)-1beta (β), IL-4, IL-6, IL-8, IL-10, complements, toll-like receptor (TLR)4, myeloid differentiation primary response gene (MYD)88, TANK-binding kinase (TBK)1, IkappaB kinase-epsilon (IKK-ε) and nuclear factor-kappaB (NF-κB) were analyzed with or without ANG treatment in tumor necrosis factor-alpha (TNF-α)- or lipopolysaccharide (LPS)-induced inflammatory HCFs by real-time polymerase chain reaction (PCR), Western blotting and immunocytochemistry. Inflammatory cytokine profiles with or without ANG were evaluated through immunodot blot analysis in inflammatory HCFs. Corneal neovascularization and opacity in a rat model of corneal alkali burn were evaluated after application of ANG eye drops.

Results

ANG decreased the mRNA levels of IL-1β, IL-6, IL-8, TNF-α receptor (TNFR)1, 2, TLR4, MYD88, and complement components except for C1r and C1s and elevated the mRNA expression of IL-4 and IL-10. Increased signal intensity of IL-6, IL-8 and monocyte chemotactic protein (MCP)-1 and MCP-2 induced by TNF-α or LPS was weakened by ANG treatment. ANG reduced the protein levels of IKK-ε by either TNF-α and LPS, and decreased TBK1 production induced by TNF-α, but not induced by LPS. The expression of NF-κB in the nuclei was decreased after ANG treatment. ANG application lowered corneal neovascularization and opacity in rats compared to controls.

Conclusion

These results demonstrate that ANG reduces the inflammatory response induced by TNF-α or LPS in HCFs through common suppression of IKK-ε-mediated activation of NF-κB. This may support the targeting of immune-mediated corneal inflammation by using ANG.
Literature
1.
Whitcher JP, Srinivasan M, Upadhyay MP. Corneal blindness: a global perspective. Bull World Health Organ. 2001;79:214–21.PubMedPubMedCentral
2.
Wang H, Zhang Y, Li Z, Wang T, Liu P. Prevalence and causes of corneal blindness. Clin Experiment Ophthalmol. 2014;42:249–53.CrossRefPubMed
3.
Xi X, McMillan DH, Lehmann GM, et al. Ocular fibroblast diversity: implications for inflammation and ocular wound healing. Invest Ophthalmol Vis Sci. 2011;52:4859–65.CrossRefPubMedPubMedCentral
4.
Liu Y, Kimura K, Yanai R, Chikama T, Nishida T. Cytokine, chemokine, and adhesion molecule expression mediated by MAPKs in human corneal fibroblasts exposed to poly(I:C). Invest Ophthalmol Vis Sci. 2008;49:3336–44.CrossRefPubMed
5.
Smith RS, Smith TJ, Blieden TM, Phipps RP. Fibroblasts as sentinel cells. Synthesis of chemokines and regulation of inflammation. Am J Pathol. 1997;151:317–22.PubMedPubMedCentral
6.
Daniels JT, Geerling G, Alexander RA, Murphy G, Khaw PT, Saarialho-Kere U. Temporal and spatial expression of matrix metalloproteinases during wound healing of human corneal tissue. Exp Eye Res. 2003;77:653–64.CrossRefPubMed
7.
Maertzdorf J, Osterhaus AD, Verjans GM. IL-17 expression in human herpetic stromal keratitis: modulatory effects on chemokine production by corneal fibroblasts. J Immunol. 2002;169:5897–903.CrossRefPubMed
8.
Mondino BJ, Sundar-Raj CV, Brady KJ. Production of first component of complement by corneal fibroblasts in tissue culture. Arch Ophthalmol. 1982;100:478–80.CrossRefPubMed
9.
Rothman B, Despins A, Webb S, et al. Cytokine regulation of C3 and C5 production by human corneal fibroblasts. Exp Eye Res. 1991;53:353–61.CrossRefPubMed
10.
11.
12.
Kawakami Y, Watanabe Y, Yamaguchi M, Sakaguchi H, Kono I, Ueki A. TNF-alpha stimulates the biosynthesis of complement C3 and factor B by human umbilical cord vein endothelial cells. Cancer Lett. 1997;116:21–6.CrossRefPubMed
13.
Nygren H, Dahlen G, Nilsson LA. Human complement activation by lipopolysaccharides from bacteroides oralis, fusobacterium nucleatum, and veillonella parvula. Infect Immun. 1979;26:391–6.PubMedPubMedCentral
14.
Sakimoto T, Yamada A, Sawa M. Release of soluble tumor necrosis factor receptor 1 from corneal epithelium by TNF-alpha-converting enzyme-dependent ectodomain shedding. Invest Ophthalmol Vis Sci. 2009;50:4618–21.CrossRefPubMed
15.
16.
Carlson EC, Drazba J, Yang X, Perez VL. Visualization and characterization of inflammatory cell recruitment and migration through the corneal stroma in endotoxin-induced keratitis. Invest Ophthalmol Vis Sci. 2006;47:241–8.CrossRefPubMed
17.
18.
Miyamoto S, Maki M, Schmitt MJ, Hatanaka M, Verma IM. Tumor necrosis factor alpha-induced phosphorylation of I kappa B alpha is a signal for its degradation but not dissociation from NF-kappa B. Proc Natl Acad Sci U S A. 1994;91:12740–4.CrossRefPubMedPubMedCentral
19.
Andreakos E, Sacre SM, Smith C, et al. Distinct pathways of LPS-induced NF-kappa B activation and cytokine production in human myeloid and nonmyeloid cells defined by selective utilization of MYD88 and Mal/TIRAP. Blood. 2004;103:2229–37.CrossRefPubMed
20.
Niederberger E, Geisslinger G. The IKK-NF-kappaB pathway: a source for novel molecular drug targets in pain therapy? FASEB J. 2008;22:3432–42.CrossRefPubMed
21.
Kravchenko VV, Mathison JC, Schwamborn K, Mercurio F, Ulevitch RJ. IKKi/IKKepsilon plays a key role in integrating signals induced by pro-inflammatory stimuli. J Biol Chem. 2003;278:26612–9.CrossRefPubMed
22.
Pomerantz JL, Baltimore D. NF-kappaB activation by a signaling complex containing TRAF2, TANK and TBK1, a novel IKK-related kinase. EMBO J. 1999;18:6694–704.CrossRefPubMedPubMedCentral
23.
24.
Lee SH, Kim KW, Min KM, Kim KW, Chang SI, Kim JC. Angiogenin reduces immune inflammation via inhibition of TANK-binding kinase 1 expression in human corneal fibroblast cells. Mediators Inflamm. 2014;2014:861435.PubMedPubMedCentral
25.
Reilly SM, Chiang SH, Decker SJ, et al. An inhibitor of the protein kinases TBK1 and IKK-varepsilon improves obesity-related metabolic dysfunctions in mice. Nat Med. 2013;19:313–21.CrossRefPubMedPubMedCentral
26.
Oikonomou KA, Kapsoritakis AN, Kapsoritaki AI, et al. Angiogenin, angiopoietin-1, angiopoietin-2, and endostatin serum levels in inflammatory bowel disease. Inflamm Bowel Dis. 2011;17:963–70.CrossRefPubMed
27.
Xu Z, Monti DM, Hu G. Angiogenin activates human umbilical artery smooth muscle cells. Biochem Biophys Res Commun. 2001;285:909–14.CrossRefPubMed
28.
Etoh T, Shibuta K, Barnard GF, Kitano S, Mori M. Angiogenin expression in human colorectal cancer: the role of focal macrophage infiltration. Clin Cancer Res. 2000;6:3545–51.PubMed
29.
Hooper LV ST, Hong CV, et al. Angiogenins: a new class of microbicidal proteins involved in innate immunity. Nat Immunol. 2003;4(3):269–73.CrossRefPubMed
30.
RA Sack LC, Krumholz D, Beaton A, et al. Membrane array characterization of 80 Chemokines, cytokines, and growth factors in open- and closed-Eye tears: angiogenin and other defense system constituents. Invest Ophthalmol Vis Sci. 2005;46(4):1228–38.CrossRef
31.
Sonoda Y, Streilein JW. Orthotopic corneal transplantation in mice--evidence that the immunogenetic rules of rejection do not apply. Transplantation. 1992;54:694–704.CrossRefPubMed
32.
33.
Srisa-Art M, Kang DK, Hong J, et al. Analysis of protein-protein interactions by using droplet-based microfluidics. Chembiochem. 2009;10:1605–11.CrossRefPubMed
34.
Fett JW, Strydom DJ, Lobb RR, et al. Isolation and characterization of angiogenin, an angiogenic protein from human carcinoma cells. Biochemistry. 1985;24:5480–6.CrossRefPubMed
35.
Moser CV, Kynast K, Baatz K, et al. The protein kinase IKKepsilon is a potential target for the treatment of inflammatory hyperalgesia. J Immunol. 2011;187:2617–25.CrossRefPubMed
36.
Cao C, Li L, Chen W, et al. Deficiency of IKKepsilon inhibits inflammation and induces cardiac protection in high-fat diet-induced obesity in mice. Int J Mol Med. 2014;34:244–52.PubMed
37.
Peant B, Diallo JS, Dufour F, et al. Over-expression of IkappaB-kinase-epsilon (IKKepsilon/IKKi) induces secretion of inflammatory cytokines in prostate cancer cell lines. Prostate. 2009;69:706–18.CrossRefPubMed
38.
Su WH, Cheng MH, Lee WL, et al. Nonsteroidal anti-inflammatory drugs for wounds: pain relief or excessive scar formation? Mediators Inflamm. 2010;2010:413238.CrossRefPubMedPubMedCentral
39.
Sonoda K, Sakamoto T, Yoshikawa H, et al. Inhibition of corneal inflammation by the topical use of Ras farnesyltransferase inhibitors: selective inhibition of macrophage localization. Invest Ophthalmol Vis Sci. 1998;39:2245–51.PubMed
40.
Nakamura T, Hamuro J, Takaishi M, et al. LRIG1 inhibits STAT3-dependent inflammation to maintain corneal homeostasis. J Clin Invest. 2014;124:385–97.CrossRefPubMed
41.
Xia Y, Feng L, Yoshimura T, Wilson CB. LPS-induced MCP-1, IL-1 beta, and TNF-alpha mRNA expression in isolated erythrocyte-perfused rat kidney. Am J Physiol. 1993;264:F774–80.PubMed
42.
Eggesbo JB, Hjermann I, Hostmark AT, Kierulf P. LPS induced release of IL-1 beta, IL-6, IL-8 and TNF-alpha in EDTA or heparin anticoagulated whole blood from persons with high or low levels of serum HDL. Cytokine. 1996;8:152–60.CrossRefPubMed
43.
44.
Grund EM, Kagan D, Tran CA, et al. Tumor necrosis factor-alpha regulates inflammatory and mesenchymal responses via mitogen-activated protein kinase kinase, p38, and nuclear factor kappaB in human endometriotic epithelial cells. Mol Pharmacol. 2008;73:1394–404.CrossRefPubMed
45.
46.
Harada A, Sekido N, Akahoshi T, Wada T, Mukaida N, Matsushima K. Essential involvement of interleukin-8 (IL-8) in acute inflammation. J Leukoc Biol. 1994;56:559–64.PubMed
47.
Ghasemi H, Ghazanfari T, Yaraee R, Faghihzadeh S, Hassan ZM. Roles of IL-8 in ocular inflammations: a review. Ocul Immunol Inflamm. 2011;19:401–12.CrossRefPubMed
48.
49.
Marie C, Pitton C, Fitting C, Cavaillon JM. Regulation by anti-inflammatory cytokines (IL-4, IL-10, IL-13, TGFbeta)of interleukin-8 production by LPS- and/or TNFalpha-activated human polymorphonuclear cells. Mediators Inflamm. 1996;5:334–40.CrossRefPubMedPubMedCentral
50.
Olson KA, Verselis SJ, Fett JW. Angiogenin is regulated in vivo as an acute phase protein. Biochem Biophys Res Commun. 1998;242:480–3.CrossRefPubMed
51.
Verselis SJ, Olson KA, Fett JW. Regulation of angiogenin expression in human HepG2 hepatoma cells by mediators of the acute-phase response. Biochem Biophys Res Commun. 1999;259:178–84.CrossRefPubMed
52.
Trouillon R, Kang DK, Park H, Chang SI, O’Hare D. Angiogenin induces nitric oxide synthesis in endothelial cells through PI-3 and Akt kinases. Biochemistry. 2010;49:3282–8.CrossRefPubMed
53.
54.
55.
Sahu A, Lambris JD. Complement inhibitors: a resurgent concept in anti-inflammatory therapeutics. Immunopharmacology. 2000;49:133–48.CrossRefPubMed
56.
Song H, Qiao F, Atkinson C, Holers VM, Tomlinson S. A complement C3 inhibitor specifically targeted to sites of complement activation effectively ameliorates collagen-induced arthritis in DBA/1J mice. J Immunol. 2007;179:7860–7.CrossRefPubMed
57.
Choi JA, Choi JS, Joo CK. Effects of amniotic membrane suspension in the rat alkali burn model. Mol Vis. 2011;17:404–12.PubMedPubMedCentral
Metadata
Title
Angiogenin ameliorates corneal opacity and neovascularization via regulating immune response in corneal fibroblasts
Authors
Seung Hoon Lee
Kyoung Woo Kim
Kwangsic Joo
Jae Chan Kim
Publication date
01-12-2016
Publisher
BioMed Central
Published in
BMC Ophthalmology / Issue 1/2016
Electronic ISSN: 1471-2415
DOI
https://doi.org/10.1186/s12886-016-0235-z

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