Correlation between pathological and functional changes of the retina and the expression of monocyte chemoattractant protein after retinal laser injury in mice_Chinese Journal of Ocular Fundus Diseases

Authors：

YuBo , ZhangXiaomin , WangMeiyan , 苏畅 , 蒋元丰 , 刘勋 ,  李筱荣

Tianjin Medical University Eye Hospital, Tianjin Medical University Eye Institute, Tianjin Medical University College of Optometry, Tianjin 300384, China;

Corresponding author：

李筱荣, Email: xiaorli@163.com

Keywords：

Retina diseases/etiology; Electroretinography; Chemokine CCL2; Animal experimentation

DOI：

10.3760/cma.j.issn.1005-1015.2014.05.017

Video：

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Objective To investigate the relationship between the pathological and functional changes of the retina and the expression of monocyte chemoattractant protein (MCP)-1 after retinal laser injury in mice. Methods A total of 116 C57BL/6 mice were randomly divided into the normal group (58 mice) and the injured group (58 mice). Retinal laser injuries were induced by Argon ion laser. At 1, 3, 7 days after laser injury, electroretinogram (ERG) responses were recorded to detect the function of the retina. Hematoxylin and eosin (HE) staining was performed to observe pathological changes. Quantitative real-time polymerase chain reaction (PCR) was performed to detect gene expression of MCP-1. Western blot was used to measure the protein expression of MCP-1. Results HE staining showed a progressive damage of the retinal structure. The results of ERG showed that the differences of dark-adaptive a wave (t=6.998, 9.594, 13.778) and b wave (t=12.089, 13.310, 21.989) amplitudes of 1, 3 and 7 day post-injury between normal group and injured group were statistically significant (P=0.000). At 1 day post-injury, the differences of light adaptive b wave amplitudes between the two groups were statistically significant (t=8.844, P=0.000). While the differences of light-adaptive a wave amplitudes were not (t=2.659,P=0.200). At 3, 7 days post-injury, the differences of a (t=3.076, 7.544) and b wave amplitudes (t=10.418, 8.485) between the two groups were statistically significant (P=0.000). In dark-adaptive ERG, the differences of a wave amplitudes between 1 day and 3 days (t=3.773), 1 day and 7 days (t=5.070) and b wave amplitudes between 1 day and 7 days (t=4.762) were statistically significant (P<0.01), while the differences of a wave amplitudes between the 3 days and 7 days (t=1.297) and b wave amplitudes between 1 day and 3 days (t=2.236), 3 day and 7 days (t=2.526) were not significant (P=0.660, 0.120, 0.060). In light-adaptive ERG, the differences of a wave amplitudes between 1 day and 7 days (t=2.992) and b wave amplitudes between 1 day and 3 days (t=3.570), 1day and 7 days (t=4.989) were statistically significant (P<0.05), while the differences of a wave amplitudes between 1 day and 3 days (t=0.516), the 3 days and 7 days (t=2.475) and b wave amplitudes between 3 days and 7 days (t=1.419) were not significant (P=1.000, 0.710, 0.070). Quantitative real-time PCR showed that the differences of MCP-1 gene expression at 1, 3 and 7 day post-injury between normal group and injured group were statistically significant (t=14.329, 16.861, 5.743; P<0.05). Western blot showed that the differences of MCP-1 protein expression at 1, 3 and 7 day post-injury between normal group and injured group were statistically significant (t=75.068, 54.145, 14.653; P<0.05). Conclusion In the first 7 days after mice retinal laser injury, there are progressive pathological and functional damage of the retina, which might be correlated with MCP-1 expression.

Citation： YuBo, ZhangXiaomin, WangMeiyan. Correlation between pathological and functional changes of the retina and the expression of monocyte chemoattractant protein after retinal laser injury in mice. Chinese Journal of Ocular Fundus Diseases, 2014, 30(5): 495-499. doi: 10.3760/cma.j.issn.1005-1015.2014.05.017 Copy

1.	Deshmane SL, Kremlev S, Amini S, et al. Monocyte chemoattractant protein-1 (MCP-1):an overview[J]. J Interferon Cytokine Res, 2009, 29:313-326.
2.	Marshall J, Bird AC. A comparative histopathological study of argon and krypton laser irradiations of the human retina[J]. Br J Ophthalmol, 1979, 63:657-668.
3.	Lambert V, Lecomte J, Hansen S, et al. Laser-induced choroidal neovascularization model to study age-related macular degeneration in mice[J]. Nat Protoc, 2013, 8:2197-2211.
4.	Ng TF, Turpie B, Masli S. Thrombospondin-1-mediated regulation of microglia activation after retinal injury[J]. Invest Ophthalmol Vis Sci, 2009, 50:5472-5478.
5.	蒋元丰,张晓敏,张琰,等. 尾静脉注射骨髓间充质干细胞对小鼠视网膜激光损伤后细胞凋亡的影响[J]. 中华眼底病杂志, 2013, 29:67-71.
6.	Belokopytov M, Shulman S, Dubinsky G, et al. Intravitreal saline injection ameliorates laser-induced retinal damage in rats[J]. Retina, 2012, 32:1165-1170.
7.	Ben-Shlomo G, Belokopytov M, Rosner M, et al. Functional deficits resulting from laser-induced damage in the rat retina[J]. Lasers Surg Med, 2006, 38:689-694.
8.	Suzuki M, Ozawa Y, Kubota S, et al. Neuroprotective response after photodynamic therapy:role of vascular endothelial growth factor[J]. J Neuroinflammation, 2011, 8:176.
9.	Kasaoka M, Ma J, Lashkari K. c-Met modulates RPE migratory response to laser-induced retinal injury. PLoS One, 2012, 7:40771.[2012-07-13]http://www.plosone.org/article/info:doi/10.1371/journal.pone.0040771?imageURI=info:doi/10.1371/journal.pone.0040771.g002.
10.	Liang KJ, Lee JE, Wang YD, et al. Regulation of dynamic behavior of retinal microglia by CX3CR1 signaling[J]. Invest Ophthalmol Vis Sci, 2009, 50:4444-4451.
11.	Cushing SD, Berliner JA, Valente AJ, et al. Minimally modified low density lipoprotein induces monocyte chemotactic protein 1 in human endothelial cells and smooth muscle cells[J]. Proc Natl Acad Sci USA, 1990, 87:5134-5138.
12.	van Coillie E, van Damme J, Opdenakker G. The MCP/eotaxin subfamily of CC chemokines[J]. Cytokine Growth Factor Rev, 1999, 10:61-86.
13.	Chen H, Liu B, Lukas TJ, et al. The aged retinal pigment epithelium/choroid:a potential substratum for the pathogenesis of age-related macular degeneration. PLoS One, 2008, 3:2339.[2008-06-04]http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.pone.0002339.
14.	Yamada K, Sakurai E, Itaya M, et al. Inhibition of laser-induced choroidal neovascularization by atorvastatin by downregulation of monocyte chemotactic protein-1 synthesis in mice[J]. Invest Ophthalmol Vis Sci, 2007, 48:1839-1843.
15.	Tsutsumi C, Sonoda KH, Egashira K, et al. The critical role of ocular-infiltrating macrophages in the development of choroidal neovascularization[J]. J Leukoc Biol, 2003, 74:25-32.
16.	Krady JK, Basu A, Allen CM, et al. Minocycline reduces proinflammatory cytokine expression, microglial activation, and caspase-3 activation in a rodent model of diabetic retinopathy[J]. Diabetes, 2005, 54:1559-1565.
17.	Dong N, Li X, Xiao L, et al. Upregulation of retinal neuronal MCP-1 in the rodent model of diabetic retinopathy and its function in vitro[J]. Invest Ophthalmol Vis Sci, 2012, 53:7567-7575.

1. Deshmane SL, Kremlev S, Amini S, et al. Monocyte chemoattractant protein-1 (MCP-1):an overview[J]. J Interferon Cytokine Res, 2009, 29:313-326.
2. Marshall J, Bird AC. A comparative histopathological study of argon and krypton laser irradiations of the human retina[J]. Br J Ophthalmol, 1979, 63:657-668.
3. Lambert V, Lecomte J, Hansen S, et al. Laser-induced choroidal neovascularization model to study age-related macular degeneration in mice[J]. Nat Protoc, 2013, 8:2197-2211.
4. Ng TF, Turpie B, Masli S. Thrombospondin-1-mediated regulation of microglia activation after retinal injury[J]. Invest Ophthalmol Vis Sci, 2009, 50:5472-5478.
5. 蒋元丰,张晓敏,张琰,等. 尾静脉注射骨髓间充质干细胞对小鼠视网膜激光损伤后细胞凋亡的影响[J]. 中华眼底病杂志, 2013, 29:67-71.
6. Belokopytov M, Shulman S, Dubinsky G, et al. Intravitreal saline injection ameliorates laser-induced retinal damage in rats[J]. Retina, 2012, 32:1165-1170.
7. Ben-Shlomo G, Belokopytov M, Rosner M, et al. Functional deficits resulting from laser-induced damage in the rat retina[J]. Lasers Surg Med, 2006, 38:689-694.
8. Suzuki M, Ozawa Y, Kubota S, et al. Neuroprotective response after photodynamic therapy:role of vascular endothelial growth factor[J]. J Neuroinflammation, 2011, 8:176.
9. Kasaoka M, Ma J, Lashkari K. c-Met modulates RPE migratory response to laser-induced retinal injury. PLoS One, 2012, 7:40771.[2012-07-13]http://www.plosone.org/article/info:doi/10.1371/journal.pone.0040771?imageURI=info:doi/10.1371/journal.pone.0040771.g002.
10. Liang KJ, Lee JE, Wang YD, et al. Regulation of dynamic behavior of retinal microglia by CX3CR1 signaling[J]. Invest Ophthalmol Vis Sci, 2009, 50:4444-4451.
11. Cushing SD, Berliner JA, Valente AJ, et al. Minimally modified low density lipoprotein induces monocyte chemotactic protein 1 in human endothelial cells and smooth muscle cells[J]. Proc Natl Acad Sci USA, 1990, 87:5134-5138.
12. van Coillie E, van Damme J, Opdenakker G. The MCP/eotaxin subfamily of CC chemokines[J]. Cytokine Growth Factor Rev, 1999, 10:61-86.
13. Chen H, Liu B, Lukas TJ, et al. The aged retinal pigment epithelium/choroid:a potential substratum for the pathogenesis of age-related macular degeneration. PLoS One, 2008, 3:2339.[2008-06-04]http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.pone.0002339.
14. Yamada K, Sakurai E, Itaya M, et al. Inhibition of laser-induced choroidal neovascularization by atorvastatin by downregulation of monocyte chemotactic protein-1 synthesis in mice[J]. Invest Ophthalmol Vis Sci, 2007, 48:1839-1843.
15. Tsutsumi C, Sonoda KH, Egashira K, et al. The critical role of ocular-infiltrating macrophages in the development of choroidal neovascularization[J]. J Leukoc Biol, 2003, 74:25-32.
16. Krady JK, Basu A, Allen CM, et al. Minocycline reduces proinflammatory cytokine expression, microglial activation, and caspase-3 activation in a rodent model of diabetic retinopathy[J]. Diabetes, 2005, 54:1559-1565.
17. Dong N, Li X, Xiao L, et al. Upregulation of retinal neuronal MCP-1 in the rodent model of diabetic retinopathy and its function in vitro[J]. Invest Ophthalmol Vis Sci, 2012, 53:7567-7575.

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Correlation between pathological and functional changes of the retina and the expression of monocyte chemoattractant protein after retinal laser injury in mice

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