Mesenchymal stem cell-derived exosomes inhibit the VEGF-A expression in human retinal vascular endothelial cells induced by high glucose

AIM: To determine the effect of exosomes derived from human umbilical cord blood mesenchymal stem cells (hUCMSCs) on the expression of vascular endothelial growth factor A (VEGF-A) in human retinal vascular endothelial cells (HRECs). METHODS: Exosomes were isolated from hUCMSCs using cryogenic ultra...

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Autores principales:	Guang-Hui He, Ying-Xue Ma, Meng Dong, Song Chen, Yu-Chuan Wang, Xiang Gao, Bin Wu, Jian Wang, Jun-Hua Wang
Formato:	article
Lenguaje:	EN
Publicado:	Press of International Journal of Ophthalmology (IJO PRESS) 2021
Materias:	mesenchymal stem cells exosomes retinal vascular endothelial cells vascular endothelial growth factor a coculture Ophthalmology RE1-994
Acceso en línea:	https://doaj.org/article/882ae91dd0274511a5f859fff9c221f0
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spelling	oai:doaj.org-article:882ae91dd0274511a5f859fff9c221f02021-11-26T04:02:12ZMesenchymal stem cell-derived exosomes inhibit the VEGF-A expression in human retinal vascular endothelial cells induced by high glucose2222-39592227-489810.18240/ijo.2021.12.03https://doaj.org/article/882ae91dd0274511a5f859fff9c221f02021-12-01T00:00:00Zhttp://ies.ijo.cn/en_publish/2021/12/20211203.pdfhttps://doaj.org/toc/2222-3959https://doaj.org/toc/2227-4898AIM: To determine the effect of exosomes derived from human umbilical cord blood mesenchymal stem cells (hUCMSCs) on the expression of vascular endothelial growth factor A (VEGF-A) in human retinal vascular endothelial cells (HRECs). METHODS: Exosomes were isolated from hUCMSCs using cryogenic ultracentrifugation and characterized by transmission electron microscopy, Western blotting and nanoparticle tracking analysis. HRECs were randomly divided into a normal control group (group A), a high glucose model group (group B), a high glucose group with 25 μg/mL (group C), 50 μg/mL (group D), and 100 μg/mL exosomes (group E). Twenty-four hours after coculture, the cell proliferation rate was detected using flow cytometry, and the VEGF-A level was detected using immunofluorescence. After coculture 8, 16, and 24h, the expression levels of VEGF-A in each group were detected using PCR and Western blots. RESULTS: The characteristic morphology (membrane structured vesicles) and size (diameter between 50 and 200 nm) were observed under transmission electron microscopy. The average diameter of 122.7 nm was discovered by nanoparticle tracking analysis (NTA). The exosomal markers CD9, CD63, and HSP70 were strongly detected. The proliferation rate of the cells in group B increased after 24h of coculture. Immunofluorescence analyses revealed that the upregulation of VEGF-A expression in HRECs stimulated by high glucose could be downregulated by cocultured hUCMSC-derived exosomes (F=39.03, P<0.01). The upregulation of VEGF-A protein (group C: F=7.96; group D: F=17.29; group E: F=11.89; 8h: F=9.45; 16h: F=12.86; 24h: F=42.28, P<0.05) and mRNA (group C: F=4.137; group D: F=13.64; group E: F=22.19; 8h: F=7.253; 16h: F=16.98; 24h: F=22.62, P<0.05) in HRECs stimulated by high glucose was downregulated by cocultured hUCMSC-derived exosomes (P<0.05). CONCLUSION: hUCMSC-derived exosomes downregulate VEGF-A expression in HRECs stimulated by high glucose in time and concentration dependent manner.Guang-Hui HeYing-Xue MaMeng DongSong ChenYu-Chuan WangXiang GaoBin WuJian WangJun-Hua WangPress of International Journal of Ophthalmology (IJO PRESS)articlemesenchymal stem cellsexosomesretinal vascular endothelial cellsvascular endothelial growth factor acocultureOphthalmologyRE1-994ENInternational Journal of Ophthalmology, Vol 14, Iss 12, Pp 1820-1827 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	mesenchymal stem cells exosomes retinal vascular endothelial cells vascular endothelial growth factor a coculture Ophthalmology RE1-994
spellingShingle	mesenchymal stem cells exosomes retinal vascular endothelial cells vascular endothelial growth factor a coculture Ophthalmology RE1-994 Guang-Hui He Ying-Xue Ma Meng Dong Song Chen Yu-Chuan Wang Xiang Gao Bin Wu Jian Wang Jun-Hua Wang Mesenchymal stem cell-derived exosomes inhibit the VEGF-A expression in human retinal vascular endothelial cells induced by high glucose
description	AIM: To determine the effect of exosomes derived from human umbilical cord blood mesenchymal stem cells (hUCMSCs) on the expression of vascular endothelial growth factor A (VEGF-A) in human retinal vascular endothelial cells (HRECs). METHODS: Exosomes were isolated from hUCMSCs using cryogenic ultracentrifugation and characterized by transmission electron microscopy, Western blotting and nanoparticle tracking analysis. HRECs were randomly divided into a normal control group (group A), a high glucose model group (group B), a high glucose group with 25 μg/mL (group C), 50 μg/mL (group D), and 100 μg/mL exosomes (group E). Twenty-four hours after coculture, the cell proliferation rate was detected using flow cytometry, and the VEGF-A level was detected using immunofluorescence. After coculture 8, 16, and 24h, the expression levels of VEGF-A in each group were detected using PCR and Western blots. RESULTS: The characteristic morphology (membrane structured vesicles) and size (diameter between 50 and 200 nm) were observed under transmission electron microscopy. The average diameter of 122.7 nm was discovered by nanoparticle tracking analysis (NTA). The exosomal markers CD9, CD63, and HSP70 were strongly detected. The proliferation rate of the cells in group B increased after 24h of coculture. Immunofluorescence analyses revealed that the upregulation of VEGF-A expression in HRECs stimulated by high glucose could be downregulated by cocultured hUCMSC-derived exosomes (F=39.03, P<0.01). The upregulation of VEGF-A protein (group C: F=7.96; group D: F=17.29; group E: F=11.89; 8h: F=9.45; 16h: F=12.86; 24h: F=42.28, P<0.05) and mRNA (group C: F=4.137; group D: F=13.64; group E: F=22.19; 8h: F=7.253; 16h: F=16.98; 24h: F=22.62, P<0.05) in HRECs stimulated by high glucose was downregulated by cocultured hUCMSC-derived exosomes (P<0.05). CONCLUSION: hUCMSC-derived exosomes downregulate VEGF-A expression in HRECs stimulated by high glucose in time and concentration dependent manner.
format	article
author	Guang-Hui He Ying-Xue Ma Meng Dong Song Chen Yu-Chuan Wang Xiang Gao Bin Wu Jian Wang Jun-Hua Wang
author_facet	Guang-Hui He Ying-Xue Ma Meng Dong Song Chen Yu-Chuan Wang Xiang Gao Bin Wu Jian Wang Jun-Hua Wang
author_sort	Guang-Hui He
title	Mesenchymal stem cell-derived exosomes inhibit the VEGF-A expression in human retinal vascular endothelial cells induced by high glucose
title_short	Mesenchymal stem cell-derived exosomes inhibit the VEGF-A expression in human retinal vascular endothelial cells induced by high glucose
title_full	Mesenchymal stem cell-derived exosomes inhibit the VEGF-A expression in human retinal vascular endothelial cells induced by high glucose
title_fullStr	Mesenchymal stem cell-derived exosomes inhibit the VEGF-A expression in human retinal vascular endothelial cells induced by high glucose
title_full_unstemmed	Mesenchymal stem cell-derived exosomes inhibit the VEGF-A expression in human retinal vascular endothelial cells induced by high glucose
title_sort	mesenchymal stem cell-derived exosomes inhibit the vegf-a expression in human retinal vascular endothelial cells induced by high glucose
publisher	Press of International Journal of Ophthalmology (IJO PRESS)
publishDate	2021
url	https://doaj.org/article/882ae91dd0274511a5f859fff9c221f0
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Mesenchymal stem cell-derived exosomes inhibit the VEGF-A expression in human retinal vascular endothelial cells induced by high glucose

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