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Effects of rat mesenchymal stem cell-derived exosomes on rat experimental autoimmune uveitis

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Author:
No author available
Journal Title:
Chinese Journal of Ocular Fundus Diseases
Issue:
6
DOI:
10.3760/cma.j.issn.1005-1015.2018.06.008
Key Word:
间质干细胞;外泌体;葡萄膜炎/治疗;动物实验;Mesenchymal stem cells;Exosomes;Uveitis/therapy;Animal experimentation

Abstract´╝Ü Objective To observe the effects of exosomes derived from rat mesenchymal stem cells (MSC-exosomes) on the rat experimental autoimmune uveitis (EAU) model.Methods Twelve Lewis rats were randomly divided into experimental group and control group by random number table,with 6 rats in each group.Rats in the experimental group were established with EAU model,100 μl of MSC-exosomes (50 μg) were periocular injected on the 9th day after modeling while the control rats were injected with the same volume of phosphate buffer.At different time points after modeling,the retinal structure was observed by hematoxylin and eosin (HE) staining,and the clinical and pathological manifestations were evaluated.T cells from the two groups were analyzed by flow cytometry,Immunohistochemical staining was used to observe the expression of macrophage surface marker CD68.The effect of MSC-exosomes on T cells was measured by lymphocyte proliferation assays.And flow cytometry was used to detect Th 1,Th 17 and regulatory T cells Variety.Electroretinogram (ERG) was used to evaluate the retinal function.Data were compared between the two groups using the t test.Results HE staining showed that the retina structure of the experimental group was more complete than that of the control group on the 15th day after modeling.Immunohistochemical staining showed that the positive expression of CD68 in the experimental group was significantly less than that in the control group.On the 15th day after modeling,the retinal pathological score of the experimental group was lower than that of the control group.On the 9th to 13th day after modeling,compared to the control group,the average clinical scores of the retina in the experimental group were lower,and the difference was statistically significant (t=3.665,3.21,3.181,4.121,3.227;P<0.01).The results of T cell proliferation assay showed that exosomes (1.0,10.0 μg/ml) inhibited the proliferation of T cells under different concentrations of R16 (1,10,30 μg/ml),and the difference was statistically significant (F=1 1.630,4.188,6.011;P<0.05).The results of flow cytometry showed that the number ofThl,Th17 and Treg cell subsets in the experimental group was decreased compared with the control group,and the difference was statistically significant (t=7.374,4.525,6.910;P<0.01).There was no difference in the proportion of cells in the T cells and lymph nodes (t=1.126,0.493,0.178;P=0.286,0.632,0.862).The results of ERG showed that,compared with the control group,the amplitudes of 0.01,3.0 cd/m2 a wave and b wave of the experiment group were all increased on the 15th day after modeling,and the differences were statistically significant (t=3.604,4.178,4.551,2.566,P<0.05).Conclusions MSC-exosomes can reduce the clinical and pathological manifestations of EAU,protect retinal function,reduce ocular macrophage infiltration,down-regulate the proportion of inflammatory cells in the eye,and inhibit T cell proliferation.

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