携带TSP-1的脐带间充质干细胞来源外泌体对视网膜静脉阻塞大鼠视网膜变性的保护作用

doi:10.3969/j.issn.1006-6233.2022.07.06

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Abstract Objective: To investigate the protective effect of human umbilical cord mesenchymal stem cell (hUCMSC)-derived exosomes carrying thrombospondin-1 (TSP-1) in a rat model of retinal vein occlusion (RVO). Methods: The hUCMSCs were randomly divided into hUCMSC group, NC-hUCMSC group and TSP-1-hUCMSC group, the NC-hUCMSC group and TSP-1-hUCMSC group were infected with the lentivirus containing TSP-1 gene sequence and the negative control lentivirus, respectively, and the cell transfection effect was determined by fluorescent inverted microscope, real-time fluorescent quantitative PCR and Western blot. The hUCMSC-derived exosomes carrying the TSP-1 gene were isolated, the morphology was observed by transmission electron microscopy, the diameter was determined by the nanoparticle tracking analysis method, and the expression of the exosomal marker proteins CD9, CD63 and Alix was detected by Western blot. The RVO model of SD rats was established by laser photodynamic method and divided into model group, hUCMSC-Exo group, TSP-1-hUCMSC-Exo group, each with 10 rats, and another 10 healthy ratswerewolf made the control group. The hUCMSC-Exo group was subconjunctivally injected with 100 μl of hUCMSC-derived exosomes, and the TSP-1-hUCMSC-Exo group was injected with 100 μL of hUCMSC-derived exosomes containing TSP-1 lentivirus infection. 1 day, 7 days, 14 days, 21 days after the establishment of the RVO model, the fundus images were observed. 21 d later, the vascular lesions were examined by fluorescein fundus angiography (FFA), and the histopathological changes of the retina were observed by HE staining, Real-time fluorescent quantitative PCR and Western blot were used to detect the mRNA and protein levels of hypoxia-inducible factor-1α (HIF-1α), vascular endothelial growth factor (VEGF) and vascular cell adhesion molecule-1 (VCAM-1) in retinal tissues. Results: The inverted fluorescence microscope showed that there was obvious green fluorescent protein expression in the cells transfected by the lentivirus, and the relative expression of TSP-1 mRNA and protein in hUCMSC in the TSP-1-hUCMSC group were significantly increased (P<0.05), indicates that the cell transfection was successful. The separated particles are spherical with a diameter of approximately 100 nm. CD9, CD63 and Alix proteins are all highly expressed, indicating that exosomes have been successfully isolated. Compared with the control group, different degrees of white occluded blood vessels and plaques appeared in the eyes of the model group on the 1st, 7th, 14th and 21th day of the model, and the model was judged to be successful; in addition, the retinal blood vessels of the model group showed tortuous and dilated veins. Uneven, fluorescence staining and exudation of blood vessel wall, retinal tissue structure was obviously damaged, and the relative expression of HIF-1α, VEGF and VCAM-1 mRNA and protein in the tissue were significantly increased (P<0.05). Compared with the model group, the hUCMSC-Exo group and TSP-1-hUCMSC-Exo group reduced the fluorescence staining of the blood vessel wall, restored the retinal structure, and the relative expression of HIF-1α, VEGF and VCAM-1 mRNA and protein decreased significantly (P<0.05). Compared with the hUCMSC-Exo group, the TSP-1-hUCMSC-Exo group did not see obvious fluorescence leakage in the blood vessels, the retina structure was clearer, and the relative expressions of HIF-1α, VEGF and VCAM-1 mRNA and protein in the tissue were significantly decreased ( P<0.05). Conclusion: HUCMSC-derived exosomes carrying TSP-1 can alleviate retinal damage in RVO rats and protect the retina. The mechanism may be related to the inhibition of the expression of HIF-1α, VEGF and VCAM-1.

Key words： Retinal vein occlusion TSP-1 Umbilical cord mesenchymal stem cell Exosomes

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	LI Xia
	ZHANG Maoju

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LI Xia,ZHANG Maoju. Protective Effect of Umbilical Cord Mesenchymal Stem Cell-Derived Exosomes Carrying TSP-1 on Retinal Degeneration in Rats with Retinal Vein Occlusion[J]. HeBei Med, 2022, 28(7): 1080-1087.

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http://www.hbyxzzs.cn/EN/10.3969/j.issn.1006-6233.2022.07.06 OR http://www.hbyxzzs.cn/EN/Y2022/V28/I7/1080

[1] Goker YS,Atilgan CU,Tekin K,et al.Association between disorganization of the retinal inner layers and capillary nonperfusion area in patients with retinal vein occlusion[J].Arq Bras Oftalmol,2020,83(6):497-504.
[2] Marcinkowska A,Cisiecki S,Rozalski M.Platelet and thrombophilia-related risk factors of retinal vein occlusion[J].Clin Med,2021,10(14):3080.
[3] Xie Q,Liu R,Jiang J,et al.What is the impact of human umbilical cord mesenchymal stem cell transplantation on clinical treatment[J].Stem Cell Res Ther,2020,11(1):519.
[4] Nikfarjam S,Rezaie J,Zolbanin NM,et al.Mesenchymal stem cell derived-exosomes:a modern approach in translational medicine[J].Transl Med,2020,18(1):449.
[5] Mir FA,Contreras-Ruiz L,Masli S.Thrombospondin-1-dependent immune regulation by transforming growth factor-β2-exposed antigen-presenting cells[J].Immunology,2015,146(4):547-556.
[6] Foulsham W,Dohlman TH,Mittal SK,et al.Thrombospondin-1 in ocular surface health and disease[J].Ocul Surf,2019,17(3):374-383.
[7] 吕小利,邹红,黎蕾,等.加味桃红四物汤灌胃治疗视网膜静脉阻塞大鼠的实验研究[J].上海中医药杂志,2019,53(8):81-88.
[8] Asato T,Nagai N,Suzuki M,et al.Closure of macular hole secondary to ischemic hemi-central retinal vein occlusion by retinal photocoagulation and topical anti-inflammatory treatment[J].Lasers Med Sci,2021,36(2):469-471.
[9] Tang Y,Zhou Y,Li HJ.Advances in mesenchymal stem cell exosomes:a review[J].Stem Cell Res Ther,2021,12(1):71.
[10] Zhang B,Wang M,Gong A,et al.HucMSC-exosome mediated-Wnt4 signaling is required for cutaneous wound healing[J].Stem Cells,2015,33(7):2158-2168.
[11] Li T,Yan Y,Wang B,et al.Exosomes derived from human umbilical cord mesenchymal stem cells alleviate liver fibrosis[J].Stem Cells Dev,2013,22(6):845-854.
[12] Suto MJ,Gupta V,Mathew B,et al.Identification of inhibitors of thrombospondin 1 activation of TGF-β[J].ACS Med Chem Lett,2020,11(6):1130-1136.
[13] Tan X,Chen Y,Foulsham W,et al.The immunoregulatory role of corneal epithelium-derived thrombospondin-1 in dry eye disease[J].Ocul Surf,2018,16(4):470-477.
[14] Blanco-Mezquita JT,Hutcheon AE,Zieske JD.Role of thrombospondin-1 in repair of penetrating corneal wounds[J].Invest Ophthalmol Vis Sci,2013,54(9):6262-6268.