Skip to main content
Top
Published in: BMC Ophthalmology 1/2017

Open Access 01-12-2017 | Research article

Assessment of biomarkers using multiplex assays in aqueous humor of patients with diabetic retinopathy

Authors: Hui Chen, Xiongze Zhang, Nanying Liao, Feng Wen

Published in: BMC Ophthalmology | Issue 1/2017

Login to get access

Abstract

Background

With the high prevalence of type 2 diabetes, diabetic retinopathy (DR) has become a leading health problem worldwide. The pathogenesis of DR is complex and several vascular, inflammatory, and neuronal mechanisms are involved. The purpose of this study was to assess the levels of immune and inflammatory biomarkers in the aqueous humor of patients with different severities of DR and to analyze the correlations between Interleukin-6 (IL-6) and these biomarkers, and between IL-6 and the severity of the disease.

Methods

Aqueous humor samples were obtained from 51 non-diabetic patients and 151 diabetic patients. Levels of 45 different cytokines, chemokines, and growth factors were measured using a multiplex bead immunoassay.

Results

IL-6, IL-8, Inducible Protein-10 (IP-10), leukemia inhibitory factor (LIF), hepatocyte growth factor (HGF) and vascular endothelial growth factor (VEGF)-A were significantly higher (p < 0.05) in the aqueous humor of the DR patients compared to the non-diabetic patients, while the concentrations of IL-1α, IL-4, IL-9, IL-21, IL-23, IL-27, IL-31, RANTES, interferon-α, growth regulated oncogene (GRO), and tumor necrosis factor (TNF)-α were significantly lower (p < 0.05) in the DR patients. The IL-6 levels increased as the severity of DR increased. In addition, the IL-6 level positively correlated with the IL-8, HGF and LIF levels, while negatively with the IL-31and GRO levels.

Conclusions

These findings suggest that inflammation and immune response may contribute to the pathogenesis of DR, and these biomarkers may potentially be new therapeutic targets for DR.
Literature
1.
go back to reference Aiello LP, Gardner TW, King GL, Blankenship G, Cavallerano JD, Ferris FL, et al. Diabetic retinopathy. Diabetes Care. 1998;21:143–56.CrossRefPubMed Aiello LP, Gardner TW, King GL, Blankenship G, Cavallerano JD, Ferris FL, et al. Diabetic retinopathy. Diabetes Care. 1998;21:143–56.CrossRefPubMed
2.
go back to reference Funatsu H, Yamashita H, Noma H, Mimura T, Nakamura S, Sakata K, et al. Aqueous humor levels of cytokines are related to vitreous levels and progression of diabetic retinopathy in diabetic patients. Graefes Arch Clin Exp Ophthalmol. 2005;243:3–8.CrossRefPubMed Funatsu H, Yamashita H, Noma H, Mimura T, Nakamura S, Sakata K, et al. Aqueous humor levels of cytokines are related to vitreous levels and progression of diabetic retinopathy in diabetic patients. Graefes Arch Clin Exp Ophthalmol. 2005;243:3–8.CrossRefPubMed
3.
go back to reference Abu el Asrar AM, Maimone D, Morse PH, Gregory S, Reder AT. Cytokines in the vitreous of patients with proliferative diabetic retinopathy. Am J Ophthalmol. 1992;114:731–6.CrossRefPubMed Abu el Asrar AM, Maimone D, Morse PH, Gregory S, Reder AT. Cytokines in the vitreous of patients with proliferative diabetic retinopathy. Am J Ophthalmol. 1992;114:731–6.CrossRefPubMed
4.
go back to reference Vignali DA. Multiplexed particle-based flow cytometric assays. J Immunol Methods. 2000;243:243–55.CrossRefPubMed Vignali DA. Multiplexed particle-based flow cytometric assays. J Immunol Methods. 2000;243:243–55.CrossRefPubMed
5.
go back to reference Sharma RK, Rogojina AT, Chalam KV. Multiplex immunoassay analysis of biomarkers in clinically accessible quantities of human aqueous humor. Mol Vis. 2009;15:60–9.PubMedPubMedCentral Sharma RK, Rogojina AT, Chalam KV. Multiplex immunoassay analysis of biomarkers in clinically accessible quantities of human aqueous humor. Mol Vis. 2009;15:60–9.PubMedPubMedCentral
6.
go back to reference Banerjee S, Savant V, Scott RA, Curnow SJ, Wallace GR, Murray PI. Multiplex bead analysis of vitreous humor of patients with vitreoretinal disorders. Invest Ophthalmol Vis Sci. 2007;48:2203–7.CrossRefPubMed Banerjee S, Savant V, Scott RA, Curnow SJ, Wallace GR, Murray PI. Multiplex bead analysis of vitreous humor of patients with vitreoretinal disorders. Invest Ophthalmol Vis Sci. 2007;48:2203–7.CrossRefPubMed
7.
go back to reference Chen H, Lin H, Zheng D, Liu Y, Chen W, Liu Y. Expression of Cytokines, Chmokines and Growth Factors in Patients Undergoing Cataract Surgery with Femtosecond Laser Pretreatment. PloS one. 2015;10(9):e0137227.CrossRefPubMedPubMedCentral Chen H, Lin H, Zheng D, Liu Y, Chen W, Liu Y. Expression of Cytokines, Chmokines and Growth Factors in Patients Undergoing Cataract Surgery with Femtosecond Laser Pretreatment. PloS one. 2015;10(9):e0137227.CrossRefPubMedPubMedCentral
8.
go back to reference Navarro-Gonzalez JF, Mora-Fernandez C. The role of inflammatory cytokines in diabetic nephropathy. J Am Soci Nephrol. 2008;19:433–42.CrossRef Navarro-Gonzalez JF, Mora-Fernandez C. The role of inflammatory cytokines in diabetic nephropathy. J Am Soci Nephrol. 2008;19:433–42.CrossRef
9.
go back to reference Oh IK, Kim SW, Oh J, Lee TS, Huh K. Inflammatory and angiogenic factors in the aqueous humor and the relationship to diabetic retinopathy. Curr Eye Res. 2010;35:1116–27.CrossRefPubMed Oh IK, Kim SW, Oh J, Lee TS, Huh K. Inflammatory and angiogenic factors in the aqueous humor and the relationship to diabetic retinopathy. Curr Eye Res. 2010;35:1116–27.CrossRefPubMed
10.
go back to reference Banks WA, Kastin AJ, Gutierrez EG. Penetration of interleukin-6 across the murine blood-brain barrier. Neurosci Lett. 1994;179:53–6.CrossRefPubMed Banks WA, Kastin AJ, Gutierrez EG. Penetration of interleukin-6 across the murine blood-brain barrier. Neurosci Lett. 1994;179:53–6.CrossRefPubMed
11.
go back to reference Moriarty AP, Spalton DJ, Moriarty BJ, Shilling JS, Ffytche TJ, Bulsara M. Studies of the blood-aqueous barrier in diabetes mellitus. Am J Ophthalmol. 1994;117:768–71.CrossRefPubMed Moriarty AP, Spalton DJ, Moriarty BJ, Shilling JS, Ffytche TJ, Bulsara M. Studies of the blood-aqueous barrier in diabetes mellitus. Am J Ophthalmol. 1994;117:768–71.CrossRefPubMed
12.
go back to reference de Vries HE, Blom-Roosemalen MC, van Oosten M, de Boer AG, van Berkel TJ, Breimer DD, et al. The influence of cytokines on the integrity of the blood-brain barrier in vitro. J Neuroimmunol. 1996;64:37–43.CrossRefPubMed de Vries HE, Blom-Roosemalen MC, van Oosten M, de Boer AG, van Berkel TJ, Breimer DD, et al. The influence of cytokines on the integrity of the blood-brain barrier in vitro. J Neuroimmunol. 1996;64:37–43.CrossRefPubMed
13.
go back to reference Funatsu H, Yamashita H, Shimizu E, Kojima R, Hori S. Relationship between vascular endothelial growth factor and interleukin-6 in diabetic retinopathy. Retina. 2001;21:469–77.CrossRefPubMed Funatsu H, Yamashita H, Shimizu E, Kojima R, Hori S. Relationship between vascular endothelial growth factor and interleukin-6 in diabetic retinopathy. Retina. 2001;21:469–77.CrossRefPubMed
14.
go back to reference Holzinger C, Weissinger E, Zuckermann A, Imhof M, Kink F, Schollhammer A, et al. Effects of interleukin-1, −2, −4, −6, interferon-gamma and granulocyte/macrophage colony stimulating factor on human vascular endothelial cells. Immunol Lett. 1993;35:109–17.CrossRefPubMed Holzinger C, Weissinger E, Zuckermann A, Imhof M, Kink F, Schollhammer A, et al. Effects of interleukin-1, −2, −4, −6, interferon-gamma and granulocyte/macrophage colony stimulating factor on human vascular endothelial cells. Immunol Lett. 1993;35:109–17.CrossRefPubMed
15.
go back to reference Funatsu H, Yamashita H, Ikeda T, Mimura T, Eguchi S, Hori S. Vitreous levels of interleukin-6 and vascular endothelial growth factor are related to diabetic macular edema. Ophthalmology. 2003;110:1690–6.CrossRefPubMed Funatsu H, Yamashita H, Ikeda T, Mimura T, Eguchi S, Hori S. Vitreous levels of interleukin-6 and vascular endothelial growth factor are related to diabetic macular edema. Ophthalmology. 2003;110:1690–6.CrossRefPubMed
16.
go back to reference Stewart CL, Kaspar P, Brunet LJ, Bhatt H, Gadi I, Kontgen F, et al. Blastocyst implantation depends on maternal expression of leukaemia inhibitory factor. Nature. 1992;359:76–9.CrossRefPubMed Stewart CL, Kaspar P, Brunet LJ, Bhatt H, Gadi I, Kontgen F, et al. Blastocyst implantation depends on maternal expression of leukaemia inhibitory factor. Nature. 1992;359:76–9.CrossRefPubMed
17.
go back to reference Kubota Y, Hirashima M, Kishi K, Stewart CL, Suda T. Leukemia inhibitory factor regulates microvessel density by modulating oxygen-dependent VEGF expression in mice. J Clin Invest. 2008;118:2393–403.PubMedPubMedCentral Kubota Y, Hirashima M, Kishi K, Stewart CL, Suda T. Leukemia inhibitory factor regulates microvessel density by modulating oxygen-dependent VEGF expression in mice. J Clin Invest. 2008;118:2393–403.PubMedPubMedCentral
18.
go back to reference Petrovic MG, Korosec P, Kosnik M, Hawlina M. Vitreous levels of interleukin-8 in patients with proliferative diabetic retinopathy. Am J Ophthalmol. 2007;143:175–6.CrossRefPubMed Petrovic MG, Korosec P, Kosnik M, Hawlina M. Vitreous levels of interleukin-8 in patients with proliferative diabetic retinopathy. Am J Ophthalmol. 2007;143:175–6.CrossRefPubMed
19.
go back to reference Loetscher M, Gerber B, Loetscher P, Jones SA, Piali L, Clark-Lewis I, et al. Chemokine receptor specific for IP10 and mig: structure, function, and expression in activated T-lymphocytes. J Exp Med. 1996;184:963–9.CrossRefPubMed Loetscher M, Gerber B, Loetscher P, Jones SA, Piali L, Clark-Lewis I, et al. Chemokine receptor specific for IP10 and mig: structure, function, and expression in activated T-lymphocytes. J Exp Med. 1996;184:963–9.CrossRefPubMed
20.
go back to reference Abu El-Asrar AM, Struyf S, Kangave D, Geboes K, Van Damme J. Chemokines in proliferative diabetic retinopathy and proliferative vitreoretinopathy. Eur Cytokine Netw. 2006;17:155–65.PubMed Abu El-Asrar AM, Struyf S, Kangave D, Geboes K, Van Damme J. Chemokines in proliferative diabetic retinopathy and proliferative vitreoretinopathy. Eur Cytokine Netw. 2006;17:155–65.PubMed
21.
go back to reference Boulday G, Haskova Z, Reinders ME, Pal S, Briscoe DM. Vascular endothelial growth factor-induced signaling pathways in endothelial cells that mediate overexpression of the chemokine IFN-gamma-inducible protein of 10 kDa in vitro and in vivo. J Immunol. 2006;176:3098–107.CrossRefPubMed Boulday G, Haskova Z, Reinders ME, Pal S, Briscoe DM. Vascular endothelial growth factor-induced signaling pathways in endothelial cells that mediate overexpression of the chemokine IFN-gamma-inducible protein of 10 kDa in vitro and in vivo. J Immunol. 2006;176:3098–107.CrossRefPubMed
22.
go back to reference Schoenberger SD, Kim SJ, Sheng J, Rezaei KA, Lalezary M, Cherney E. Increased prostaglandin E2 (PGE2) levels in proliferative diabetic retinopathy, and correlation with VEGF and inflammatory cytokines. Investi Ophthalmol Vis Sci. 2012;53:5906–11.CrossRef Schoenberger SD, Kim SJ, Sheng J, Rezaei KA, Lalezary M, Cherney E. Increased prostaglandin E2 (PGE2) levels in proliferative diabetic retinopathy, and correlation with VEGF and inflammatory cytokines. Investi Ophthalmol Vis Sci. 2012;53:5906–11.CrossRef
23.
go back to reference Lange CA, Stavrakas P, Luhmann UF, de Silva DJ, Ali RR, Gregor ZJ, et al. Intraocular oxygen distribution in advanced proliferative diabetic retinopathy. Am J Ophthalmol. 2011;152:406–12.CrossRefPubMed Lange CA, Stavrakas P, Luhmann UF, de Silva DJ, Ali RR, Gregor ZJ, et al. Intraocular oxygen distribution in advanced proliferative diabetic retinopathy. Am J Ophthalmol. 2011;152:406–12.CrossRefPubMed
24.
go back to reference Crane IJ, Kuppner MC, McKillop-Smith S, Knott RM, Forrester JV. Cytokine regulation of RANTES production by human retinal pigment epithelial cells. Cell Immunol. 1998;184:37–44.CrossRefPubMed Crane IJ, Kuppner MC, McKillop-Smith S, Knott RM, Forrester JV. Cytokine regulation of RANTES production by human retinal pigment epithelial cells. Cell Immunol. 1998;184:37–44.CrossRefPubMed
25.
go back to reference Meleth AD, Agron E, Chan CC, Reed GF, Arora K, Byrnes G, et al. Serum inflammatory markers in diabetic retinopathy. Investi Ophthalmol Vis Sci. 2005;46:4295–301.CrossRef Meleth AD, Agron E, Chan CC, Reed GF, Arora K, Byrnes G, et al. Serum inflammatory markers in diabetic retinopathy. Investi Ophthalmol Vis Sci. 2005;46:4295–301.CrossRef
26.
go back to reference Ozturk BT, Bozkurt B, Kerimoglu H, Okka M, Kamis U, Gunduz K. Effect of serum cytokines and VEGF levels on diabetic retinopathy and macular thickness. Mol Vis. 2009;15:1906–14.PubMedPubMedCentral Ozturk BT, Bozkurt B, Kerimoglu H, Okka M, Kamis U, Gunduz K. Effect of serum cytokines and VEGF levels on diabetic retinopathy and macular thickness. Mol Vis. 2009;15:1906–14.PubMedPubMedCentral
27.
go back to reference Adamis AP, Miller JW, Bernal MT, D'Amico DJ, Folkman J, Yeo TK, et al. Increased vascular endothelial growth factor levels in the vitreous of eyes with proliferative diabetic retinopathy. Am J Ophthalmol. 1994;118:445–50.CrossRefPubMed Adamis AP, Miller JW, Bernal MT, D'Amico DJ, Folkman J, Yeo TK, et al. Increased vascular endothelial growth factor levels in the vitreous of eyes with proliferative diabetic retinopathy. Am J Ophthalmol. 1994;118:445–50.CrossRefPubMed
28.
go back to reference Duh E, Aiello LP. Vascular endothelial growth factor and diabetes: the agonist versus antagonist paradox. Diabetes. 1999;48:1899–906.CrossRefPubMed Duh E, Aiello LP. Vascular endothelial growth factor and diabetes: the agonist versus antagonist paradox. Diabetes. 1999;48:1899–906.CrossRefPubMed
29.
go back to reference Sydorova M, Lee MS. Vascular endothelial growth factor levels in vitreous and serum of patients with either proliferative diabetic retinopathy or proliferative vitreoretinopathy. Ophthalmic Res. 2005;37:188–90.CrossRefPubMed Sydorova M, Lee MS. Vascular endothelial growth factor levels in vitreous and serum of patients with either proliferative diabetic retinopathy or proliferative vitreoretinopathy. Ophthalmic Res. 2005;37:188–90.CrossRefPubMed
30.
go back to reference Bhargava M, Joseph A, Knesel J, Halaban R, Li Y, Pang S, et al. Scatter factor and hepatocyte growth factor: activities, properties, and mechanism. Cell Growth Differ. 1992;3:11–20.PubMed Bhargava M, Joseph A, Knesel J, Halaban R, Li Y, Pang S, et al. Scatter factor and hepatocyte growth factor: activities, properties, and mechanism. Cell Growth Differ. 1992;3:11–20.PubMed
31.
go back to reference Grierson I, Heathcote L, Hiscott P, Hogg P, Briggs M, Hagan S. Hepatocyte growth factor/scatter factor in the eye. Prog Retin Eye Res. 2000;19:779–802.CrossRefPubMed Grierson I, Heathcote L, Hiscott P, Hogg P, Briggs M, Hagan S. Hepatocyte growth factor/scatter factor in the eye. Prog Retin Eye Res. 2000;19:779–802.CrossRefPubMed
32.
go back to reference Canton A, Burgos R, Hernandez C, Mateo C, Segura RM, Mesa J, et al. Hepatocyte growth factor in vitreous and serum from patients with proliferative diabetic retinopathy. Br J Ophthalmol. 2000;84:732–5.CrossRefPubMedPubMedCentral Canton A, Burgos R, Hernandez C, Mateo C, Segura RM, Mesa J, et al. Hepatocyte growth factor in vitreous and serum from patients with proliferative diabetic retinopathy. Br J Ophthalmol. 2000;84:732–5.CrossRefPubMedPubMedCentral
33.
go back to reference Simo R, Vidal MT, Garcia-Arumi J, Carrasco E, Garcia-Ramirez M, Segura RM, et al. Intravitreous hepatocyte growth factor in patients with proliferative diabetic retinopathy: a case-control study. Diabetes Res Clin Pract. 2006;71:36–44.CrossRefPubMed Simo R, Vidal MT, Garcia-Arumi J, Carrasco E, Garcia-Ramirez M, Segura RM, et al. Intravitreous hepatocyte growth factor in patients with proliferative diabetic retinopathy: a case-control study. Diabetes Res Clin Pract. 2006;71:36–44.CrossRefPubMed
34.
go back to reference Leibovitch I, Loewenstein A, Alster Y, Rosenblatt I, Lazar M, Yassur Y, et al. Interferon alpha-2a for proliferative diabetic retinopathy after complete laser panretinal photocoagulation treatment. Ophthalmic Surg Lasers Imaging. 2004;35:16–22.PubMed Leibovitch I, Loewenstein A, Alster Y, Rosenblatt I, Lazar M, Yassur Y, et al. Interferon alpha-2a for proliferative diabetic retinopathy after complete laser panretinal photocoagulation treatment. Ophthalmic Surg Lasers Imaging. 2004;35:16–22.PubMed
35.
go back to reference Skowsky WR, Siddiqui T, Hodgetts D, Lambrou FH Jr, Stewart MW, Foster MT Jr. A pilot study of chronic recombinant interferon-alfa 2a for diabetic proliferative retinopathy: metabolic effects and opthalmologic effects. J Diabetes Complicat. 1996;10:94–9.CrossRefPubMed Skowsky WR, Siddiqui T, Hodgetts D, Lambrou FH Jr, Stewart MW, Foster MT Jr. A pilot study of chronic recombinant interferon-alfa 2a for diabetic proliferative retinopathy: metabolic effects and opthalmologic effects. J Diabetes Complicat. 1996;10:94–9.CrossRefPubMed
Metadata
Title
Assessment of biomarkers using multiplex assays in aqueous humor of patients with diabetic retinopathy
Authors
Hui Chen
Xiongze Zhang
Nanying Liao
Feng Wen
Publication date
01-12-2017
Publisher
BioMed Central
Published in
BMC Ophthalmology / Issue 1/2017
Electronic ISSN: 1471-2415
DOI
https://doi.org/10.1186/s12886-017-0572-6

Other articles of this Issue 1/2017

BMC Ophthalmology 1/2017 Go to the issue