The effect of netrin-1 on the retinal vascular permeability in early diabetic retinopathy rats_Chinese Journal of Ocular Fundus Diseases

Authors：

ZhouXianhui ,  MengXuxia , SunYunbo , 胡迭 , 郭庆敏

Department of Ophthalmology, The Affiliated Hospital of Qingdao University, Qingdao 266500, China;

Corresponding author：

MengXuxia, Email: mengxuxia@163.com

Keywords：

Diabetic retinopathy/physiopathology; Blood retinal barrier/physiopathology; Nerve growth factors; Animal experimentation

DOI：

10.3760/cma.j.issn.1005-1015.2015.05.015

Video：

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Objective To explore the effect and mechanism of netrin-1 on blood-retinal barrier permeability in diabetes mellitus (DM) rats. Methods Eighty Sprague-Dawley rats were randomly divided into the normal control group, DM+balanced salt solution (BSS) group, DM+netrin-1 low dose group and DM+netrin-1 high dose group, with 20 rats in each group. DM rats were induced by intraperitoneal injection of streptozocin (STZ). These rats were feed with high sugar and fat for 3 months after STZ injection. All rats were sacrificed at 1 month after intravitreal injection. Retinal vascular permeability was measured by Evans blue. The expression level of occludin was determined by immunohistochemistry. Hematoxylin-eosin (HE) staining of retina was used to observe the pathological change of DM and the level of occludin mRNA was analyzed by real-time fluorescence quantitative reverse transcription polymerase chain reaction (RT-PCR). Five rats of each group. Results HE staining of retina showed that the degree of edema and vascularization in DM+netrin-1 high dose group was better than DM+BSS group. Staining of occludin in retina was limited to nerve fiber layer, ganglion cells, inner plexiform layer and inner nuclear layer in normal rats, but in DM+BSS group, the color of staining positive of occludin was lighter and more reduced. However, DM+ netrin-1 group occludin staining was deepen and enlarged. The result of RT-PCR showed that the expression of occludin mRNA in other three groups was less than normal control group (P < 0.05). The significant difference during DM+BSS group, low dose group and DM+netrin-1 high dose group (F=177.13, P=0.00), and the more concentrate of netrin-1 the higher expression of occluding. Compared the DM+netrin-1 low dose group with DM+BSS group, there was significant difference expression of occludin (t=-13.98, P=0.00). There was significant difference between the DM+netrin-1 high dose group and normal control group (t=12.87, P=0.00). There was statistically significant difference in DM+BSS group, DM+netrin-1 low dose group and DM+netrin-1 high dose group (F=179.69, P=0.00). Compared the two group of different concentration netrin-1, the quantification of vascular permeability in DM+netrin-1 high dose group reduced more (t=12.73, P=0.00). Conclusions Netrin-1 can protect the blood-retinal barrier in DM rats. Netrin-1 may decrease BRB leakage in DM rats by protecting the expression of occludin.

Citation： ZhouXianhui, MengXuxia, SunYunbo. The effect of netrin-1 on the retinal vascular permeability in early diabetic retinopathy rats. Chinese Journal of Ocular Fundus Diseases, 2015, 31(5): 472-476. doi: 10.3760/cma.j.issn.1005-1015.2015.05.015 Copy

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1. Kollias AN, Ulbig MW. Diabetic retinopathy:early diagnosis and effective treatment[J]. Dtsch Arztebl Int, 2010, 107(5):75-78.
2. Liang B, Jin ML, Liu HB. Water-soluble polysaccharide from dried lycium barb arum fruits: isolation, structural feature Sandantioxi dan activity carbohydrate polymers[J]. Ophthmology, 2011, 83:1947-1951.
3. Tessier-Lavigne M, Goodman CS.The molecular biology of axon guidance [J].Science, 1996, 274(5290):1123-1133.
4. Park KW, Crouse D, Lee M, et al. The axonal attractant Netrin-1 is an angiogenic factor.[J].Proc Natl Acad Sci USA, 2004, 101(46):16210-16215.
5. 谢涵, 邹丽, 朱剑文, 等.轴突导向因子netrin-1基因沉默对胎盘血管生成影响的体内外实验研究[J].中华妇产科杂志, 2011, 46(5):364-369.
6. Vinores SA, Van NIEL E, Swerdioff JL, et al. Electron microscopic immunocytochemical demonstration of blood retinal barrier breakdown in human diabetics and its association with aldose reductase in retinal vascular endothelium and pigment epithelium[J]. Histochem J, 1993, 25(9):648-663.
7. Takenaga Y, Takagi N, Murotomi K, et al. Inhibition of Src activity decreases tyrosine phosphorylation of occludin in brain capillaries and attenuates increase in permeability of the blood-brain barrier after transient focal cerebral ischemia[J].J Cereb Blood Flow Metab, 2009, 29(6):1099-1108.
8. Förster C.Tight junctions and the modulation of barrier function in disease[J]. Histochem Cell Biol, 2008, 130(1):55-70.
9. Larrivee B, Freitas C, Suchting S, et al. Guidance of vascular development: lessons from the nervous system[J].Circ Res, 2009, 104(4):428-441.
10. Nguyen A, Cai H.Netrin-1 induces angiogenesis via a DCC-dependent ERK1/2-eNOS feed-forward mechanism[J].Proc Natl Acad Sci USA, 2006, 103(17):6530-5653.
11. Fan Y, Shen F, Chen Y, et al. Overexpression of netrin-1 induces neovascularization in the adult mouse brain.[J].J Cereb Blood Flow Metab, 2008, 28(9): 1543-1551.
12. Lu X, Le Noble F, Yuan L, et al.The netrin receptor UNC5B mediates guidance events controlling morphogenesis of the vascular system[J]. Nature, 2004, 432, 179-186.
13. Bouvrée K, Larrivée B, Lv X, et al.Netrin-1 inhibits sprouting angiogenesis in developing avian embryos[J].Dev Biol, 2008, 318(1):172-183.
14. Yang Y, Zou L, Wang Y, et al. Axon guidance cue netrin-1 has dual function in angiogenesis[J].Cancer Biol Ther, 2007, 6(5):743-748.
15. Zhang X, Liu J, Xiong S, et al.Expression of netrin-1 in diabetic rat retina[J].Eye Sci, 2013, 28(3)148-152.

Chinese Journal of Ocular Fundus Diseases

The effect of netrin-1 on the retinal vascular permeability in early diabetic retinopathy rats

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