The effect of interleukin 10 modified endothelial progenitor cells in rat diabetic retinopathy_Chinese Journal of Ocular Fundus Diseases

Authors：

WangYing , JiangFeng , ChenQingzhong , 张竹红 , 孟祥达 ,  颜华

Department of Ophthalmology, General Hospital of Tianjin Medical University, Tianjin 300052, China;

Corresponding author：

颜华, Email: phuayan2000@163.com

Keywords：

Diabetic retinopathy/physiopathology; Interleukin-10; Stem cells; Animal experimentation

DOI：

10.3760/cma.j.issn.1005-1015.2015.06.016

Video：

Export PDF Favorites Scan Get Citation

Abstract Full text Figures/Tables Video References Cited by

Objective To observe the effect of interleukin (IL) 10 modified endothelial progenitor cells (EPC) in diabetic retinopathy (DR). Methods EPC cells were collected and cultivated from the bone marrow of rats and identified by immuno-fluorescence staining. EPC cells were infected with lentivirus (LV) of EPC-LV-IL10-GFP (EPC-LV-IL10-GFP group) or EPC-LV-NC-GFP (GFP group). EPC cells without lentivirus infection was the EPC group. Enzyme-linked immuno sorbent assay (ELISA) was used to measure the concentrations of tumor necrosis factor (TNF)-α, IL10, IL8 and vascular endothelial growth factor (VEGF) in the supernatant of these three groups. 168 male Wistar rats were divided into normal control group (28 rats), diabetes mellitus (DM) group (28 rats), DM-blank control group (56 rats) and DM-intervention group (56 rats). DM was introduced in the latter 3 groups by streptozotocin intravenous injection. Three months later, the rats in the DM-blank control group and DM-intervention group were injected with EPC-LV-NC-GFP or EPC-LV-IL10-GFP by tail vein, respectively. Immunohistochemistry was used to observe the GFP expression in rat retinas. The blood-retinal barrier breakdown was detected by Evans blue (EB) dye. The retinal histopathologic changes were observed by transmission electron microscope. The mRNA level of VEGF, matrix metallproteinases-9 (MMP-9), angiopoietin-1 (Ang-1), inducible nitric oxide synthase (iNOS) and endothelial nitric oxide synthase (eNOS) in retina were measured by reverse transcription-polymerase chain reaction (RT-PCR). Results ELISA showed that the levels of TNF-αand IL8 in the supernatant significantly decreased, while the levels of IL10 and VEGF increased (P < 0.05) in EPC-LV-IL10-GFP group. GFP expressed in the retina of blank control group and intervention group, mainly in the ganglion cell layer, inner nuclear layer and outer plexiform layer. The retinal blood vessel pathological change and EB permeability significantly decreased in intervention group compared with DM group (P < 0.05), and blank control group (P < 0.05). RT-PCR revealed that the mRNA level of VEGF, MMP-9 and Ang-1 significantly increased, and eNOS decreased in DM group compared to the normal control group (P < 0.05). The mRNA level of VEGF and iNOS decreased, eNOS increased while Ang-1 and MMP-9 had not changed in DM-blank control group and DM-intervention group compared with DM group (P < 0.05). Conclusions IL10 modified EPC can improve the inflammative microenvironment and suppressed the pathogenesis of DR. Furthermore, EPC transplantation can increase the number of EPC and exerted their effect.

Citation： WangYing, JiangFeng, ChenQingzhong. The effect of interleukin 10 modified endothelial progenitor cells in rat diabetic retinopathy. Chinese Journal of Ocular Fundus Diseases, 2015, 31(6): 575-580. doi: 10.3760/cma.j.issn.1005-1015.2015.06.016 Copy

1.	Zhang W, Yan H. Simvastatin increases circulating endothelial progenitor cells and reduces the formation and progression of diabetic retinopathy in rats[J]. Exp Eye Res, 2012, 105:1-8.
2.	Zhang W, Yan H. Dysfunction of circulating endothelial progenitor cells in type 1 diabetic rats with diabetic retinopathy[J]. Graefe's Arch Clin Exp Ophthalmol, 2013, 251(4):1123-1131.
3.	Atesok K, Li R, Stewart DJ, et al. Endothelial progenitor cells promote fracture healing in a segmental bone defect model[J]. J Orthop Res, 2010, 28(8):1007-1014.
4.	Algandaby MM, Alghamdi HA, Ashour OM, et al. Mechanisms of the antihyperglycemic activity of Retamaraetam in streptozotocin-induced diabetic rats[J]. Food Chem Toxicol, 2010, 48(8-9): 2448-2453.
5.	Xu Q, Qaum T, Adamis AP. Sensitive blood-retinal barrier breakdown quantitation using Evans blue[J]. Invest Ophthalmol Vis Sci, 2001, 42(3):789-794.
6.	Chen J, Chen S. Transfusion of CXCR4-primed endothelial progenitor cells reduces cerebral ischemic damage and promotes repair in db/db diabetic mice[J/OL]. PLoS One, 2012, 7(11):50105[2012-11-21].http://dx.plos.org/10.1371/journal.pone.0050105.
7.	Varma NR, Janic B, Iskander AS, et al. Endothelial progenitor cells (EPCs) as gene carrier system for rat model of human glioma[J/OL]. PLos One, 2012, 7(1):30310[2012-01-20].http://dx.plos.org/10.1371/journal.pone.0030310.
8.	Krishnamurthy P, Thal M, Verma S, et al. Interleukin-10 deficiency impairs bone marrow-derived endothelial progenitor cell survival and function in ischemic myocardium[J]. Circ Res, 2011, 109(11): 1280-1289.
9.	Joussen AM, Poulaki V, Tsujikawa A, et al. Suppression of diabetic retinopathy with angiopoietin-1[J]. Am J Pathol, 2002, 160(5):1683-1693.
10.	Hattori K, Dias S, Heissig B, et al. Vascular endothelial growth factor and angiopoietin-1 stimulate postnatal hematopoiesis by recruitment of vasculogenic and hematopoietic stem cells[J]. J Exp Med, 2001, 193(9):1005-1014.
11.	Kowluru RA, Zhong Q, Santos JM. Matrix metalloproteinases in diabetic retinopathy: potential role of MMP-9[J]. Expert Opin Investig Drugs, 2012, 21(6):797-805.
12.	Hfistov M, Erl R, Weber PC. Endothelial progenitor cells mobilization, differentiation, and homing[J]. Arterioscler Thromb Vasc Biol, 2003, 23(7):1185-1189.
13.	Mishra A, Newman EA. Inhibition of inducible nitric oxide synthase reverses the loss of functional hyperemia in diabetic retinopathy[J]. Glia, 2010, 58(16):1996-2004.

1. Zhang W, Yan H. Simvastatin increases circulating endothelial progenitor cells and reduces the formation and progression of diabetic retinopathy in rats[J]. Exp Eye Res, 2012, 105:1-8.
2. Zhang W, Yan H. Dysfunction of circulating endothelial progenitor cells in type 1 diabetic rats with diabetic retinopathy[J]. Graefe's Arch Clin Exp Ophthalmol, 2013, 251(4):1123-1131.
3. Atesok K, Li R, Stewart DJ, et al. Endothelial progenitor cells promote fracture healing in a segmental bone defect model[J]. J Orthop Res, 2010, 28(8):1007-1014.
4. Algandaby MM, Alghamdi HA, Ashour OM, et al. Mechanisms of the antihyperglycemic activity of Retamaraetam in streptozotocin-induced diabetic rats[J]. Food Chem Toxicol, 2010, 48(8-9): 2448-2453.
5. Xu Q, Qaum T, Adamis AP. Sensitive blood-retinal barrier breakdown quantitation using Evans blue[J]. Invest Ophthalmol Vis Sci, 2001, 42(3):789-794.
6. Chen J, Chen S. Transfusion of CXCR4-primed endothelial progenitor cells reduces cerebral ischemic damage and promotes repair in db/db diabetic mice[J/OL]. PLoS One, 2012, 7(11):50105[2012-11-21].http://dx.plos.org/10.1371/journal.pone.0050105.
7. Varma NR, Janic B, Iskander AS, et al. Endothelial progenitor cells (EPCs) as gene carrier system for rat model of human glioma[J/OL]. PLos One, 2012, 7(1):30310[2012-01-20].http://dx.plos.org/10.1371/journal.pone.0030310.
8. Krishnamurthy P, Thal M, Verma S, et al. Interleukin-10 deficiency impairs bone marrow-derived endothelial progenitor cell survival and function in ischemic myocardium[J]. Circ Res, 2011, 109(11): 1280-1289.
9. Joussen AM, Poulaki V, Tsujikawa A, et al. Suppression of diabetic retinopathy with angiopoietin-1[J]. Am J Pathol, 2002, 160(5):1683-1693.
10. Hattori K, Dias S, Heissig B, et al. Vascular endothelial growth factor and angiopoietin-1 stimulate postnatal hematopoiesis by recruitment of vasculogenic and hematopoietic stem cells[J]. J Exp Med, 2001, 193(9):1005-1014.
11. Kowluru RA, Zhong Q, Santos JM. Matrix metalloproteinases in diabetic retinopathy: potential role of MMP-9[J]. Expert Opin Investig Drugs, 2012, 21(6):797-805.
12. Hfistov M, Erl R, Weber PC. Endothelial progenitor cells mobilization, differentiation, and homing[J]. Arterioscler Thromb Vasc Biol, 2003, 23(7):1185-1189.
13. Mishra A, Newman EA. Inhibition of inducible nitric oxide synthase reverses the loss of functional hyperemia in diabetic retinopathy[J]. Glia, 2010, 58(16):1996-2004.

Chinese Journal of Ocular Fundus Diseases

The effect of interleukin 10 modified endothelial progenitor cells in rat diabetic retinopathy

Abstract Full text Figures/Tables Video References Cited by

Previous Article

Next Article

Format

Content