Expression of Toll-like receptor 4 and inflammatory factors in retina of diabetic rats_Chinese Journal of Ocular Fundus Diseases

Authors：

YuShujing ,  WangKang , LiNa , 林鹏耀 , 李茜 , 李明铭

Department of Ophthalmology, Beijing Friendship Hospital, Beijing Capital Medical University, Beijing 100050, China;

Corresponding author：

WangKang, Email: bnbn2000@sohu.com

Keywords：

Diabetic retinopathy/pathophysiology; Toll-like receptor 4/immunology; Drosophila proteins/immunology

DOI：

10.3760/cma.j.issn.1005-1015.2014.02.014

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Objective To observe the expression of Toll-like receptor 4 (TLR4) and inflammatory cytokines, leucocytic density and permeability in retina of diabetic rat. Methods A total of 106 Brown Norway rats were randomly divided into experimental group and control group with 53 rats in each group. Diabetic model was established in experimental group by intraperitoneal injection of streptozotocin, and control rats received intraperitoneal injection of an equal volume of citric acid-sodium citrate buffer. Four weeks later, the retinas were collected for further analysis. TLR4 RNA and protein expression were measured by quantitative polymerase chain reaction and Western blot. Inflammatory cytokines, including tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), monocyte chemo-attractant protein-1 (MCP-1), were measured by enzyme-linked immunosorbent assay in rat retina homogenate. Leukocyte density in the retina was measured by acridine orange fundus angiography. The retinal permeability was evaluated by Evans blue (EB) staining. Results TLR4 expression was significantly increased in diabetic rats of experimental group compared with non-diabetic rats of control group (F=1.606, 0.789; P < 0.05). Inflammatory cytokines (TNF-α, IL-1β and MCP-1) were significantly increased in retina of diabetic rats of experimental group versus non-diabetic rat of control group (F=24.622, 5.758, 4.829; P < 0.05). The retinal leukocyte density was (6.2±0.5)×10^-5, (2.2±0.3)×10^-5 cells/pixel² in experimental and control group respectively, the difference was statistically significant (F=2.025, P < 0.05). The amount of retinal EB leakage was (23.41±4.47), (13.22±3.59) ng/mg in experimental and control group respectively, the difference was statistically significant (F=21.08, P < 0.05). Conclusion TLR4 and inflammatory cytokines expression, leucocytic density and permeability increased significantly in retina of diabetic rat.

Citation： YuShujing, WangKang, LiNa. Expression of Toll-like receptor 4 and inflammatory factors in retina of diabetic rats. Chinese Journal of Ocular Fundus Diseases, 2014, 30(2): 175-179. doi: 10.3760/cma.j.issn.1005-1015.2014.02.014 Copy

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2.	Biondi-Zoccai GG, Abbate A, Liuzzo G, et al. Atherothrombosis, inflammation, and diabetes[J]. J Am Coll Cardiol, 2003, 41:1071-1077.
3.	Medzhitov R. Toll-like receptors and innate immunity[J]. Nat Rev Immunol, 2001, 1:135-145.
4.	Devaraj S, Tobias P, Jialal I. Knockout of toll-like receptor-4 attenuates the pro-inflammatory state of diabetes[J]. Cytokine, 2011, 55:441-445.
5.	Crook M. Type 2 diabetes mellitus:a disease of the innate immune system an update[J]. Diabet Med, 2004, 21:203-207.
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7.	Buraczynska M, Baranowicz-Gaszczyk I, Tarach J, et al. Toll-like receptor 4 gene polymorphism and early onset of diabetic retinopathy in patients with type 2 diabetes[J]. Hum Immunol, 2009, 70:121-124.
8.	Yamamoto M, Sato S, Hemmi H, et al. Role of adaptor TRIF in the MyD88-independent toll-like receptor signaling pathway[J]. Science, 2003, 301:640-643.
9.	Mohammad MK, Morran M, Slotterbeck B, et al. Dysregulated toll-like receptor expression and signaling in bone marrow-derived macrophages at the onset of diabetes in the non-obese diabetic mouse[J]. Int Immunol, 2006, 18:1101-1113.
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13.	王康, 王艳玲, 高立新, 等.地塞米松对糖尿病大鼠视网膜血管内白细胞聚集、血管通透性及细胞间黏附因子-1表达的影响[J].中华眼底病杂志, 2007, 23:273-276.
14.	Joussen AM, Poulaki V, Qin W, et al. Retinal vascular endothelial growth factor induces intercellular adhesion molecule-1 and endothelial nitric oxide synthase expression and initiates early diabetic retinal leukocyte adhesion in vivo[J].Am J Pathol, 2002, 160:501-509.
15.	Miyahara S, Kiryu J, Yamashiro K, et al. Simvastatin inhibits leukocyte accumulation and vascular permeability in the retinas of rats with streptozotocin-induced diabetes[J]. Am J Pathol, 2004, 164:1697-1706.
16.	Nishiwaki H, Ogura Y, Kimura H, et al. Quantitative evaluation of leukocyte dynamics in retinal microcirculation[J]. Invest Ophthalmol Vis Sci, 1995, 36:123-130.
17.	Miyamoto K, Khosrof S, Bursell SE, et al. Prevention of leukostasis and vascular leakage in streptozotocin-induced diabetic retinopathy via intercellular adhesion molecule-1 inhibition[J]. Proc Natl Acad Sci USA, 1999, 96:10836-10841.
18.	Joussen AM, Poulaki V, Mitsiades N, et al. Suppression of Fas-FasL-induced endothelial cell apoptosis prevents diabetic blood-retinal barrier breakdown in a model of streptozotocin induced diabetes[J]. FASEB J, 2003, 17:76-78.
19.	Miyamoto K, Khosrof S, Bursell SE, et al. Vascular endothelial growth factor (VEGF)-induced retinal vascular permeability is mediated by intercellular adhesion molecule-1 (ICAM-1)[J]. Am J Pathol, 2000, 156:1733-1739.
20.	Joussen AM, Murata T, Tsujikawa A, et al. Leukocyte-mediated endothelial cell injury and death in the diabetic retina[J]. Am J Pathol, 2001, 158:147-152.
21.	Kim I, Moon SO, Kim SH, et al. Vascular endothelial growth factor expression of intercellular adhesion molecule 1 (ICAM-1), vascular cell adhesion molecule 1 (VCAM-1), and E-selectin through nuclear factor-kappa B activation in endothelial cells[J]. J Biol Chem, 2001, 276:7614-7620.

1. Joussen AM, Poulaki V, Le ML, et al. A central role for inflammation in the pathogenesis of diabetic retinopathy[J].FASEB J, 2004, 18:1450-1452.
2. Biondi-Zoccai GG, Abbate A, Liuzzo G, et al. Atherothrombosis, inflammation, and diabetes[J]. J Am Coll Cardiol, 2003, 41:1071-1077.
3. Medzhitov R. Toll-like receptors and innate immunity[J]. Nat Rev Immunol, 2001, 1:135-145.
4. Devaraj S, Tobias P, Jialal I. Knockout of toll-like receptor-4 attenuates the pro-inflammatory state of diabetes[J]. Cytokine, 2011, 55:441-445.
5. Crook M. Type 2 diabetes mellitus:a disease of the innate immune system an update[J]. Diabet Med, 2004, 21:203-207.
6. Kolb H, Mandrup-Poulsen T. An immune origin of type 2 diabetes?[J]. Diabetologia, 2005, 48:1038-1050.
7. Buraczynska M, Baranowicz-Gaszczyk I, Tarach J, et al. Toll-like receptor 4 gene polymorphism and early onset of diabetic retinopathy in patients with type 2 diabetes[J]. Hum Immunol, 2009, 70:121-124.
8. Yamamoto M, Sato S, Hemmi H, et al. Role of adaptor TRIF in the MyD88-independent toll-like receptor signaling pathway[J]. Science, 2003, 301:640-643.
9. Mohammad MK, Morran M, Slotterbeck B, et al. Dysregulated toll-like receptor expression and signaling in bone marrow-derived macrophages at the onset of diabetes in the non-obese diabetic mouse[J]. Int Immunol, 2006, 18:1101-1113.
10. Devaraj S, Dasu MR, Park SH, et al. Increased levels of ligands of Toll-like receptors 2 and 4 in type 1 diabetes[J]. Diabetologia, 2009, 52:1665-1668.
11. Dasu MR, Devaraj S, Zhao L, et al. High glucose induces toll-like receptor expression in human monocytes:mechanism of activation[J]. Diabetes, 2008, 57:3090-3098.
12. Hodgkinson CP, Laxton RC, Patel K, et al. Advanced glycation end-product of low density lipoprotein activates the toll-like 4 receptor pathway implications for diabetic atherosclerosis[J]. Arterioscler Thromb Vasc Biol, 2008, 28:2275-2281.
13. 王康, 王艳玲, 高立新, 等.地塞米松对糖尿病大鼠视网膜血管内白细胞聚集、血管通透性及细胞间黏附因子-1表达的影响[J].中华眼底病杂志, 2007, 23:273-276.
14. Joussen AM, Poulaki V, Qin W, et al. Retinal vascular endothelial growth factor induces intercellular adhesion molecule-1 and endothelial nitric oxide synthase expression and initiates early diabetic retinal leukocyte adhesion in vivo[J].Am J Pathol, 2002, 160:501-509.
15. Miyahara S, Kiryu J, Yamashiro K, et al. Simvastatin inhibits leukocyte accumulation and vascular permeability in the retinas of rats with streptozotocin-induced diabetes[J]. Am J Pathol, 2004, 164:1697-1706.
16. Nishiwaki H, Ogura Y, Kimura H, et al. Quantitative evaluation of leukocyte dynamics in retinal microcirculation[J]. Invest Ophthalmol Vis Sci, 1995, 36:123-130.
17. Miyamoto K, Khosrof S, Bursell SE, et al. Prevention of leukostasis and vascular leakage in streptozotocin-induced diabetic retinopathy via intercellular adhesion molecule-1 inhibition[J]. Proc Natl Acad Sci USA, 1999, 96:10836-10841.
18. Joussen AM, Poulaki V, Mitsiades N, et al. Suppression of Fas-FasL-induced endothelial cell apoptosis prevents diabetic blood-retinal barrier breakdown in a model of streptozotocin induced diabetes[J]. FASEB J, 2003, 17:76-78.
19. Miyamoto K, Khosrof S, Bursell SE, et al. Vascular endothelial growth factor (VEGF)-induced retinal vascular permeability is mediated by intercellular adhesion molecule-1 (ICAM-1)[J]. Am J Pathol, 2000, 156:1733-1739.
20. Joussen AM, Murata T, Tsujikawa A, et al. Leukocyte-mediated endothelial cell injury and death in the diabetic retina[J]. Am J Pathol, 2001, 158:147-152.
21. Kim I, Moon SO, Kim SH, et al. Vascular endothelial growth factor expression of intercellular adhesion molecule 1 (ICAM-1), vascular cell adhesion molecule 1 (VCAM-1), and E-selectin through nuclear factor-kappa B activation in endothelial cells[J]. J Biol Chem, 2001, 276:7614-7620.

Chinese Journal of Ocular Fundus Diseases

Expression of Toll-like receptor 4 and inflammatory factors in retina of diabetic rats

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